US20030236208A1

US20030236208A1 - Targeted chromosomal genomic alterations in plants using modified single stranded oligonucleotides

Info

Publication number: US20030236208A1
Application number: US10/307,005
Authority: US
Inventors: Eric Kmiec; Howard Gamper; Michael Rice; Jungsup Kim
Original assignee: University of Delaware
Current assignee: University of Delaware
Priority date: 2000-06-01
Filing date: 2002-11-26
Publication date: 2003-12-25
Also published as: WO2001092512A2; AU2001265277B2; WO2001092512A3; EP1297122A2; US7258854B2; US20030217377A1; AU6527701A; US6936467B2; IL153122A0; US20030051270A1; CA2410523A1

Abstract

Presented are methods and compositions for targeted chromosomal genomic alterations with modified single-stranded oligonucleotides. The oligonucleotides of the invention have modified nuclease-resistant termini comprising LNA, phosphorothioate linkages or 2′-O-Me base analogues or combinations of such modifications.

Description

FIELD OF THE INVENTION

The technical field of the invention is oligonucleotide-directed repair or alteration of plant genetic information using novel chemically modified oligonucleotides.

BACKGROUND OF THE INVENTION

A number of methods have been developed specifically to alter the genomic information of plants. These methods generally include the use of vectors such as, for example, T-DNA, carrying nucleic acid sequences encoding partial or complete portions of a particular protein which is expressed in a cell or tissue to effect the alteration. The expression of the particular protein then results in the desired phenotype. See, for example, U.S. Pat. No. 4,459,355 which describes a method for transforming plants with a DNA vector and U.S. Pat. No. 5,188,642 which describes cloning or expression vectors containing a transgenic DNA sequence which when expressed in plants confers resistance to the herbicide glyphosate. The use of such transgene-containing vectors adds one or more exogenous copies of a gene in a usually random fashion at one or more integration sites of the plant's genome at some variable frequency. The introduced gene may be foreign or may be derived from the host plant. Any gene which was originally present in the genome, which may be, for example, a normal allelic variant, mutated, defective, and/or functional copy of the introduced gene, is retained in the genome of the host plant.

These methods of gene alteration are problematic in that complications which can compromise the vigor, productivity, yield, etc. of the plant may result. One such problem is that insertion of exogenous nucleic acid at random location(s) in the genome can have deleterious effects. The random nature of this insertion and/or the use of exogenous promoters can also cause the timing, location or strength of expression of the introduced transgene to be inappropriate or unpredictable. Another problem with such systems includes the addition of unnecessary and unwanted genetic material to the genome of the recipient, including, for example, T-DNA ends or other vector remnants, exogenous control sequences required to allow production of the transgene protein, which control sequences may be exogenous or native to the host plant and/or the transgene, and reporter genes or resistance markers. Such remnants and added sequences may have presently unrecognized consequences, for example, involving genetic rearrangements of the recipient genomes. In addition, concerns have been raised with consumption, especially by humans, of plants containing such exogenous genetic material.

More recently, simpler systems involving poly- or oligo-nucleotides have been described for use in the alteration of genomic DNA. These chimeric RNA-DNA oligonucleotides, requiring contiguous RNA and DNA bases in a double-stranded molecule folded by complementarity into a double hairpin conformation, have been shown to effect single basepair or frameshift alterations, for example, for mutation or repair of plant, animal or fungal genomes. See, for example, WO 99/07865 and U.S. Pat. No. 5,565,350. In the chimeric RNA-DNA oligonucleotide, an uninterrupted stretch of DNA bases within the molecule is required for sequence alteration of the targeted genome while the obligate RNA residues are involved in complex stability. Due to the length, backbone composition, and structural configuration of these chimeric RNA-DNA molecules, they are expensive to synthesize and difficult to purify. Moreover, if the RNA-containing strand of the chimeric RNA-DNA oligonucleotide is designed so as to direct gene alteration, a series of mutagenic reactions resulting in nonspecific base alteration can result. Such a result reduces the utility of such a molecule in methods designed for targeted gene alteration.

Alternatively, other oligo- or poly-nucleotides have been used which require a triplex forming, usually polypurine or polypyrimidine, structural domain which binds to a DNA helical duplex through Hoogsteen interactions between the major groove of the DNA duplex and the oligonucleotide. Such oligonucleotides may have an additional DNA reactive moiety, such as psoralen, covalently linked to the oligonucleotide. These reactive moieties function as effective intercalation agents, stabilize the formation of a triplex and can be mutagenic. Such agents may be required in order to stabilize the triplex forming domain of the oligonucleotide with the DNA double helix if the Hoogsteen interactions from the oligonucleotide/target base composition are insufficient. See, e.g., U.S. Pat. No. 5,422,251. The utility of these oligonucleotides for directing targeted gene alteration is compromised by a high frequency of nonspecific base changes.

In more recent work, the domain for altering a genome is linked or tethered to the triplex forming domain of the bi-functional oligonucleotide, adding an additional linking or tethering functional domain to the oligonucleotide. See, e.g., Culver et al., Nature Biotechnology 17: 989-93 (1999). Such chimeric or triplex forming molecules have distinct structural requirements for each of the different domains of the complete poly- or oligo-nucleotide in order to effect the desired genomic alteration in either episomal or chromosomal targets.

Other genes, e.g. CFTR, have been targeted by homologous recombination using duplex fragments having several hundred basepairs. See, e.g., Kunzelmann et al., Gene Ther. 3:859-867 (1996). Similar efforts to target genes by homologous recombination in plants using large fragments of DNA had some success. See Kempin et al., Nature 389:802-803 (1997). However, the efficiency and reproducibility of the published homologous recombination approach in plants has severely limited the widespread use of this method.

Earlier experiments to mutagenize an antibiotic resistance indicator gene by homologous recombination used an unmodified DNA oligonucleotide rather than larger fragments of DNA, wherein the oligonucleotide had no functional domains other than a region of complementary sequence to the target. See Campbell et al., New Biologist 1: 223-227 (1989). These experiments required large concentrations of the oligonucleotide, exhibited a very low frequency of episomal modification of a targeted exogenous plasmid gene not normally found in the cell and have not been reproduced. However, as shown in examples herein, we have observed that an unmodified DNA oligonucleotide can convert a base at low frequency which is detectable using the assay systems described herein.

Oligonucleotides designed for use in the targeted alteration of genetic information are significantly different from oligonucleotides designed for antisense approaches. For example, antisense oligonucleotides are perfectly complementary to and bind an mRNA strand in order to modify expression of a targeted mRNA and are used at high concentration. As a consequence, they are unable to produce a gene conversion event by either mutagenesis or repair of a defect in the chromosomal DNA of a host genome. Furthermore, the backbone chemical composition used in most oligonucleotides designed for use in antisense approaches renders them inactive as substrates for homologous pairing or mismatch repair enzymes and the high concentrations of oligonucleotide required for antisense applications can be toxic with some types of nucleotide modifications. In addition, antisense oligonucleotides must be complementary to the mRNA and therefore, may not be complementary to the other DNA strand or to genomic sequences that span the junction between intron sequence and exon sequence.

Artificial chromosomes can be useful for the screening purposes identified herein. These molecules are man-made linear or circular DNA molecules constructed from essential cis-acting DNA sequence elements that are responsible for the proper replication and partitioning of natural chromosomes (Murray et al., 1983). The essential elements are: (1) Autonomous Replication Sequences (ARS), (2) Centromeres, and (3) Telomeres.

Yeast artificial chromosomes (YACs) allow large segments of genomic DNA to be cloned and modified (Burke et al., Science 236:806; Peterson et al., Trends Genet. 13:61 (1997); Choi, et al., Nat. Genet., 4:117-223 (1993), Davies, et al., Biotechnology 11:911-914 (1993), Matsuura, et al., Hum. Mol. Genet., 5:451-459 (1996), Peterson et al., Proc. Natl. Acad. Sci., 93:6605-6609 (1996); and Schedl, et al., Cell, 86:71-82 (1996)). Other vectors also have been developed for the cloning of large segments of genomic DNA, including cosmids, and bacteriophage P1 (Sternberg et al., Proc. Natl. Acad. Sci. U.S.A., 87:103-107 (1990)). YACs have certain advantages over these alternative large capacity cloning vectors (Burke et al., Science, 236:806-812 (1987)). The maximum insert size is 35-30 kb for cosmids, and 100 kb for bacteriophage P1, both of which are much smaller than the maximal insert size for a YAC.

An alternative to YACs are cloning systems based on the E. coli fertility factor that have been developed to construct large genomic DNA insert libraries. They are bacterial artificial chromosomes (BACs) and P-1 derived artificial chromosomes (PACs) (Mejia et al., Genome Res. 7:179-186 (1997); Shizuya et al., Proc. Natl. Acad. Sci. 89:8794-8797 (1992); Ioannou et al., Nat. Genet., 6:84-89 (1994); Hosoda et al., Nucleic Acids Res. 18:3863 (1990)). BACs are based on the E. coli fertility plasmid (F factor); and PACs are based on the bacteriophage P1. These vectors propagate at a very low copy number (1-2 per cell) enabling genomic inserts up to 300 kb in size to be stably maintained in recombination deficient hosts. The PACs and BACs are circular DNA molecules that are readily isolated from the host genomic background by classical alkaline lysis (Birnboim et al., Nucleic Acids Res. 7:1513-1523 (1979)). In addition, BACs have been developed for transformation of plants with high-molecular weight DNA using the T-DNA system (Hamilton, Gene 24:107-116 (1997); Frary & Hamilton, Transgenic Res. 10: 121-132 (2001)).

A need exists for simple, inexpensive oligonucleotides capable of producing targeted alteration of genetic material such as those described herein as well as methods to identify optimal oligonucleotides that accurately and efficiently alter target DNA.

SUMMARY OF THE INVENTION

Novel, modified single-stranded nucleic acid molecules that direct gene alteration in plants are identified and the efficiency of alteration is analyzed both in vitro using a cell-free extract assay and in vivo using a yeast system and a plant system. The alteration in an oligonucleotide of the invention may comprise an insertion, deletion, substitution, as well as any combination of these. Site specific alteration of DNA is not only useful for studying function of proteins in vivo, but it is also useful for creating plants with desired phenotypes, including, for example, environmental stress tolerance, improved nutritional value, herbicide resistance, disease resistance, modified oil production, modified starch production, and altered floral morphology including selective sterility. As described herein, oligonucleotides of the invention target directed specific gene alterations in genomic double-stranded DNA in cells. The target genomic DNA can be nuclear chromosomal DNA as well as plastid or mitochondrial chromosomal DNA. The target DNA can also be a transgene present in the plant cell, including, for example, a previously introduced T-DNA. For screening purposes, the target plant DNA can also be extrachromosomal DNA present in plant or non-plant cells in various forms including, e.g., mammalian artificial chromosomes (MACs), PACs from P-1 vectors, yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), plant artificial chromosomes (PLACs), as well as episomal DNA, including episomal DNA from an exogenous source such as a plasmid or recombinant vector. Many of these artificial chromosome constructs containing plant DNA can be obtained from a variety of sources, including, e.g., the Arabidopsis Biological Resource Center (ABRC) at the Ohio State University, and the Rice Genome Research Program at the MAFF DNA bank in Ibaraki, Japan. The target DNA may be transcriptionally silent or active. In a preferred embodiment, the target DNA to be altered is the non-transcribed strand of a genomic DNA duplex. In a more preferred embodiment, the target DNA to be altered is the non-transcribed strand of a transcribed gene of a genomic DNA duplex.

The low efficiency of targeted gene alteration obtained using unmodified DNA oligonucleotides is believed to be largely the result of degradation by nucleases present in the reaction mixture or the target cell. Although different modifications are known to have different effects on the nuclease resistance of oligonucleotides or stability of duplexes formed by such oligonucleotides (see, e.g., Koshkin et al., J. Am. Chem. Soc., 120:13252-3), we have found that it is not possible to predict which of any particular known modification would be most useful for any given alteration event, including for the construction of gene alteration oligonucleotides, because of the interaction of different as yet unidentified proteins during the gene alteration event. Herein, a variety of nucleic acid analogs have been developed that increase the nuclease resistance of oligonucleotides that contain them, including, e.g., nucleotides containing phosphorothioate linkages or 2′-O-methyl analogs. We recently discovered that single-stranded DNA oligonucleotides modified to contain 2′-O-methyl RNA nucleotides or phosphorothioate linkages can enable specific alteration of genetic information at a higher level than either unmodified single-stranded DNA or a chimeric RNA/DNA molecule. See, for example, copending applications U.S. application Ser. No. 60/208,538, U.S. application Ser. No. 60/244,989, U.S. application Ser. No. 09/818,875, international application no. PCT/US01/09761 and Gamper et al., Nucleic Acids Research 28: 4332-4339 (2000), the disclosures of which are incorporated herein in their entirety by reference. We also found that additional nucleic acid analogs which increase the nuclease resistance of oligonucleotides that contain them, including, e.g., “locked nucleic acids” or “LNAs”, xylo-LNAs and L-ribo-LNAs; see, for example, Wengel & Nielsen, WO 99/14226; Wengel, WO 00/56748; Wengel, WO 00/66604; and Jakobsen & Koshkin, WO 01/25478 also allow specific targeted alteration of genetic information.

The assay allows for determining the optimum length of the oligonucleotide, optimum sequence of the oligonucleotide, optimum position of the mismatched base or bases, optimum chemical modification or modifications, optimum strand targeted for identifying and selecting the most efficient oligonucleotide for a particular gene alteration event by comparing to a control oligonucleotide. Control oligonucleotides may include a chimeric RNA-DNA double hairpin oligonucleotide directing the same gene alteration event, an oligonucleotide that matches its target completely, an oligonucleotide in which all linkages are phosphorothiolated, an oligonucleotide fully substituted with 2′-O-methyl analogs or an RNA oligonucleotide. Such control oligonucleotides either fail to direct a targeted alteration or do so at a lower efficiency as compared to the oligonucleotides of the invention. The assay further allows for determining the optimum position of a gene alteration event within an oligonucleotide, optimum concentration of the selected oligonucleotide for maximum alteration efficiency by systematically testing a range of concentrations, as well as optimization of either the source of cell extract by testing different plants or strains, or testing cells derived from different plants or strains, or plant cell lines. Using a series of single-stranded oligonucleotides, comprising all RNA or DNA residues and various mixtures of the two, several new structures are identified as viable molecules in nucleotide conversion to direct or repair a genomic mutagenic event. When extracts from mammalian, plant and fungal cells are used and are analyzed using a genetic readout assay in bacteria, single-stranded oligonucleotides having one of several modifications are found to be more active than a control RNA-DNA double hairpin chimera structure when evaluated using an in vitro gene repair assay. Similar results are also observed in vivo using yeast, mammalian and plant cells. Molecules containing various lengths of modified bases were found to possess greater activity than unmodified single-stranded DNA molecules.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides oligonucleotides having chemically modified, nuclease resistant residues, preferably at or near the termini of the oligonucleotides, and methods for their identification and use in targeted alteration of plant genetic material, including gene mutation, targeted gene repair and gene knockout. The oligonucleotides are preferably used for mismatch repair or alteration by changing at least one nucleic acid base, or for frameshift repair or alteration by addition or deletion of at least one nucleic acid base. The oligonucleotides of the invention direct any such alteration, including gene correction, gene repair or gene mutation and can be used, for example, to introduce a polymorphism or haplotype or to eliminate (“knockout”) a particular protein activity. For example, gene alterations that knockout a particular protein activity can be obtained using oligonucleotides designed to convert a codon in the coding region of the protein to a stop codon, thus prematurely terminating translation of the protein. Oligonucleotides that introduce stop codons in the open-reading-frame of the protein are one embodiment of the invention. Generally, oligonucleotides that introduce stop codons early in the open-reading-frame of the protein are preferred. If the open-reading-frame contains more than one methionine, oligonucleotides that introduce stop codons after the second methionine are preferred. Additionally, if the gene exhibits alternative splice sites, oligonucleotides that introduce stop codons in exons after the alternative splice site are preferred. The following table provides examples of codons that can be converted to stop codons by altering a single oligonucleotide. A skilled artisan could readily identify other codons that can be converted to stop codons by altering one, two or three of the base pairs in a given codon. Similarly, a skilled artisan could readily identify codons that can be converted to stop codons by a frameshift mutations that inserts or deletes one or two base pairs in the open-reading-frame. It is also understood that more than one stop codon can be generated in a single open-reading-frame and that these stop codons can be adjacent in the sequence or separated by intervening codons. Where more than one stop codon is introduced into a single open-reading-frame, such alterations can be generated by a single or multiple oligonucleotides and can be generated simultaneously or by sequential mutagenesis of the target nucleic acid.



	Corresponding
Original codons*	stop codon

G GA (glycine), A GA (arginine), C GA (arginine), T T A	TGA
(leucine), T C A (serine), TG T (cysteine), TG G
(tryptophan), TG C (cysteine)
A AG (lysine), G AG (glutamate), C AG (glutamine), T T G	TAG
(leucine), T C G (serine), T G G (tryptophan), TA T
(cysteine), TA C (tyrosine)
A AA (lysine), G AA (glutamate), C AA (glutamine), T T A	TAA
(leucine), T C A (serine), TA T (cysteine), TA C
(tyrosine)

The oligonucleotides of the invention are designed as substrates for homologous pairing and repair enzymes and as such have a unique backbone composition that differs from chimeric RNA-DNA double hairpin oligonucleotides, antisense oligonucleotides, and/or other poly- or oligo-nucleotides used for altering genomic DNA, such as triplex forming oligonucleotides. The single-stranded oligo-nucleotides described herein are inexpensive to synthesize and easy to purify. In side-by-side comparisons, an optimized single-stranded oligonucleotide comprising modified residues as described herein is significantly more efficient than a chimeric RNA-DNA double hairpin oligonucleotide in directing a base substitution or frameshift mutation in a cell-free extract assay.

We have discovered that single-stranded oligonucleotides having a DNA domain surrounding the targeted base, with the domain preferably central to the poly- or oligo-nucleotide, and having at least one modified end, preferably at the 3′ terminal region, are able to alter a target genetic sequence and with an efficiency that is higher than chimeric RNA-DNA double hairpin oligonucleotides disclosed in U.S. Pat. No. 5,565,350. Preferred oligonucleotides of the invention have at least two modified bases on at least one of the termini, preferably the 3′ terminus of the oligonucleotide. Oligonucleotides of the invention can efficiently be used to introduce targeted alterations in a genetic sequence of DNA in the presence of human, animal, plant, fungal (including yeast) proteins and in cells of different types including, for example, plant cells, fungal cells including S. cerevisiae, Ustillago maydis, Candida albicans, and mammalian cells. Particularly preferred are cells and cell extracts derived from plants including, for example, experimental model plants such as Chiamydomonas reinhardtii, Physcomitrella patens, and Arabidopsis thaliana in addition to crop plants such as cauliflower (Brassica oleracea), artichoke (Cynara scolymus), fruits such as apples (Malus, e.g. domesticus), mangoes (Mangifera, e.g. indica), banana (Musa, e.g. acuminata), berries (such as currant, Ribes, e.g. rubrum), kiwifruit (Actinidia, e.g. chinensis), grapes (Vitis, e.g. vinifera), bell peppers (Capsicum, e.g. annuum), cherries (such as the sweet cherry, Prunus, e.g. avium), cucumber (Cucumis, e.g. sativus), melons (Cucumis, e.g. melo), nuts (such as walnut, Juglans, e.g. regia; peanut, Arachis hypogeae), orange (Citrus, e.g. maxima), peach (Prunus, e.g. persica), pear (Pyra, e.g. communis), plum (Prunus, e.g. domestica), strawberry (Fragaria, e.g. moschata or vesca), tomato (Lycopersicon, e.g. esculentum); leaves and forage, such as alfalfa (Medicago, e.g. sativa or truncatula), cabbage (e.g. Brassica oleracea), endive (Cichoreum, e.g. endivia), leek (Allium, e.g. porrum), lettuce (Lactuca, e.g. sativa), spinach (Spinacia, e.g. oleraceae), tobacco (Nicotiana, e.g. tabacum); roots, such as arrowroot (Maranta, e.g. arundinacea), beet (Beta, e.g. vulgaris), carrot (Daucus, e.g. carota), cassava (Manihot, e.g. esculenta), turnip (Brassica, e.g. rapa), radish (Raphanus, e.g. sativus), yam (Dioscorea, e.g. esculenta), sweet potato (Ipomoea batatas); seeds, including oilseeds, such as beans (Phaseolus, e.g. vulgaris), pea (Pisum, e.g. sativum), soybean (Glycine, e.g. max), cowpea (Vigna unguiculata), mothbean (Vigna aconitifolia), wheat (Triticum, e.g. aestivum), sorghum (Sorghum e.g. bicolor), barley (Hordeum, e.g. vulgare), corn (Zea, e.g. mays), rice (Oryza, e.g. sativa), rapeseed (Brassica napus), millet (Panicum sp.), sunflower (Helianthus annuus), oats (Avena sativa), chickpea (Cicer, e.g. arietinum); tubers, such as kohlrabi (Brassica, e.g. oleraceae), potato (Solanum, e.g. tuberosum) and the like; fiber and wood plants, such as flax (Linum e.g. usitatissimum), cotton (Gossypium e.g. hirsutum), pine (Pinus sp.), oak (Quercus sp.), eucalyptus (Eucalyptus sp.), and the like and ornamental plants such as turfgrass (Lolium, e.g. rigidum), petunia (Petunia, e.g. x hybrida), hyacinth (Hyacinthus orientalis), carnation (Dianthus e.g. caryophyllus), delphinium (Delphinium, e.g. ajacis), Job's tears (Coix lacryma-jobi), snapdragon (Antirrhinum majus), poppy (Papaver, e.g. nudicaule), lilac (Syringa, e.g. vulgaris), hydrangea (Hydrangea e.g. macrophylla), roses (including Gallicas, Albas, Damasks, Damask Perpetuals, Centifolias, Chinas, Teas and Hybrid Teas) and ornamental goldenrods (e.g. Solidago spp.). Such plant cells can then be used to regenerate whole plants according to methods described herein or any method known in the art. The DNA domain of the oligonucleotides is preferably fully complementary to one strand of the gene target, except for the mismatch base or bases responsible for the gene alteration event(s). On either side of the preferably central DNA domain, the contiguous bases may be either RNA bases or, preferably, are primarily DNA bases. The central DNA domain is generally at least 8 nucleotides in length. The base(s) targeted for alteration in the most preferred embodiments are at least about 8, 9 or 10 bases from one end of the oligonucleotide.

According to certain embodiments, one or both of the termini of the oligonucleotides of the present invention comprise phosphorothioate modifications, LNA backbone (including LNA derivatives and analogs) modifications, or 2′-O-methyl base analogs, or any combination of these modifications. Oligonucleotides comprising 2′-O-methyl or LNA analogs are a mixed DNA/RNA polymer. The oligonucleotides of the invention are, however, single-stranded and are not designed to form a stable internal duplex structure within the oligonucleotide. The efficiency of gene alteration is surprisingly increased with oligonucleotides having internal complementary sequence comprising phosphorothioate modified bases as compared to 2′-O-methyl modifications. This result indicates that specific chemical interactions are involved between the converting oligonucleotide and the proteins involved in the conversion. The effect of other such chemical interactions to produce nuclease resistant termini using modifications other than LNA (including LNA derivatives or analogs), phosphorothioate linkages, or 2′-O-methyl analog incorporation into an oligonucleotide can not yet be predicted because the proteins involved in the alteration process and their particular chemical interaction with the oligonucleotide substituents are not yet known and cannot be predicted.

In the examples, oligonucleotides of defined sequence are provided for alteration of genes in particular plants. Provided the teachings of the instant application, one of skill in the art could readily design oligonucleotides to introduce analogous alterations in homologous genes from any plant. Furthermore, in the tables of these examples, the oligonucleotides of the invention are not limited to the particular sequences disclosed. The oligonucleotides of the invention include extensions of the appropriate sequence of the longer 120 base oligonucleotides which can be added base by base to the smallest disclosed oligonucleotides of 17 bases. Thus the oligonucleotides of the invention include for each correcting change, oligonucleotides of

length

17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, or 120 with further single-nucleotide additions up to the longest sequence disclosed. In some embodiments, longer nucleic acids of up to 240 bases which comprise the sequences disclosed herein may be used. Moreover, the oligonucleotides of the invention do not require a symmetrical extension on either side of the central DNA domain. Similarly, the oligonucleotides of the invention as disclosed in the various tables for alteration of particular plant genes contain phosphorothioate linkages, 2′-O-methyl analog or LNA (including LNA derivatives and analogs) or any combination of these modifications just as the assay oligonucleotides do.

The present invention, however, is not limited to oligonucleotides that contain any particular nuclease resistant modification. Oligonucleotides of the invention may be altered with any combination of additional LNAs (including LNA derivatives and analogs), phosphorothioate linkages or 2′-O-methyl analogs to maximize conversion efficiency. For oligonucleotides of the invention that are longer than about 17 to about 25 bases in length, internal as well as terminal region segments of the backbone may be altered. Alternatively, simple fold-back structures at each end of a oligonucleotide or appended end groups may be used in addition to a modified backbone for conferring additional nuclease resistance.

The different oligonucleotides of the present invention preferably contain more than one of the aforementioned backbone modifications at each end. In some embodiments, the backbone modifications are adjacent to one another. However, the optimal number and placement of backbone modifications for any individual oligonucleotide will vary with the length of the oligonucleotide and the particular type of backbone modification(s) that are used. If constructs of identical sequence having phosphorothioate linkages are compared, 2, 3, 4, 5, or 6 phosphorothioate linkages at each end are preferred. If constructs of identical sequence having 2′-O-methyl base analogs are compared, 1, 2, 3 or 4 analogs are preferred. The optimal number and type of backbone modifications for any particular oligo-nucleotide useful for altering target DNA may be determined empirically by comparing the alteration efficiency of the oligonucleotide comprising any combination of the modifications to a control molecule of comparable sequence using any of the assays described herein. The optimal position(s) for oligonucleotide modifications for a maximally efficient altering oligonucleotide can be determined by testing the various modifications as compared to control molecule of comparable sequence in one of the assays disclosed herein. In such assays, a control molecule includes, e.g., a completely 2′-O-methyl substituted molecule, a completely complementary oligonucleotide, or a chimeric RNA-DNA double hairpin.

Increasing the number of phosphorothioate linkages, LNAs or 2′-O-methyl bases beyond the preferred number generally decreases the gene repair activity of a 25 nucleotide long oligonucleotide. Based on analysis of the concentration of oligonucleotide present in the extract after different time periods of incubation, it is believed that the terminal modifications impart nuclease resistance to the oligo-nucleotide thereby allowing it to survive within the cellular environment. However, this may not be the only possible mechanism by which such modifications confer greater efficiency of conversion. For example, as disclosed herein, certain modifications to oligonucleotides confer a greater improvement to the efficiency of conversion than other modifications.

Efficiency of conversion is defined herein as the percentage of recovered substrate molecules that have undergone a conversion event. Depending on the nature of the target genetic material, e.g. the genome of a cell, efficiency could be represented as the proportion of cells or clones containing an extrachromosomal element that exhibit a particular phenotype. Alternatively, representative samples of the target genetic material can be sequenced to determine the percentage that have acquired the desire change. The oligonucleotides of the invention in different embodiments can alter DNA two, three, four, five, six, seven, eight, nine, ten, twelve, fifteen, twenty, thirty, and fifty or more fold more than control oligonucleotides. Such control oligonucleotides are oligonucleotides with fully phosphorothiolated linkages, oligonucleotides that are fully substituted with 2′-O-methyl analogs, a perfectly matched oligonucleotide that is fully complementary to a target sequence or a chimeric DNA-RNA double hairpin oligonucleotide such as disclosed in U.S. Pat. No. 5,565,350.

In addition, for a given oligonucleotide length, additional modifications interfere with the ability of the oligonucleotide to act in concert with the cellular recombination or repair enzyme machinery which is necessary and required to mediate a targeted substitution, addition or deletion event in DNA. For example, fully phosphorothiolated or fully 2-O-methylated molecules are inefficient in targeted gene alteration.

The oligonucleotides of the invention as optimized for the purpose of targeted alteration of genetic material, including gene knockout or repair, are different in structure from antisense oligo-nucleotides that may possess a similar mixed chemical composition backbone. The oligonucleotides of the invention differ from such antisense oligonucleotides in chemical composition, structure, sequence, and in their ability to alter genomic DNA. Significantly, antisense oligonucleotides fail to direct targeted gene alteration. The oligonucleotides of the invention may target either strand of DNA and can include any component of the genome including, for example, intron and exon sequences. The preferred embodiment of the invention is a modified oligonucleotide that binds to the non-transcribed strand of a genomic DNA duplex. In other words, the preferred oligonucleotides of the invention target the sense strand of the DNA, i.e. the oligonucleotides of the invention are complementary to the non-transcribed strand of the target duplex DNA. The sequence of the non-transcribed strand of a DNA duplex is found in the mRNA produced from that duplex, given that mRNA uses uracil-containing nucleotides in place of thymine-containing nucleotides.

Moreover, the initial observation that single-stranded oligonucleotides comprising these modifications and lacking any particular triplex forming domain have reproducibly enhanced gene alteration activity in a variety of assay systems as compared to a chimeric RNA-DNA double-stranded hairpin control or single-stranded oligonucleotides comprising other backbone modifications was surprising. The single-stranded molecules of the invention totally lack the complementary RNA binding structure that stabilizes a normal chimeric double-stranded hairpin of the type disclosed in U.S. Pat. No. 5,565,350 yet is more effective in producing targeted base conversion as compared to such a chimeric RNA-DNA double-stranded hairpin. In addition, the molecules of the invention lack any particular triplex forming domain involved in Hoogsteen interactions with the DNA double helix and required by other known oligonucleotides in other oligonucleotide-dependant gene conversion systems. Although the lack of these functional domains was expected to decrease the efficiency of an alteration in a sequence, just the opposite occurs: the efficiency of sequence alteration using the modified oligonucleotides of the invention is higher than the efficiency of sequence alteration using a chimeric RNA-DNA hairpin targeting the same sequence alteration. Moreover, the efficiency of sequence alteration or gene conversion directed by an unmodified oligonucleotide is many times lower as compared to a control chimeric RNA-DNA molecule or the modified oligonucleotides of the invention targeting the same sequence alteration. Similarly, molecules containing at least 3 2′-O-methyl base analogs are about four to five fold less efficient as compared to an oligonucleotide having the same number of phosphorothioate linkages.

The oligonucleotides of the present invention for alteration of a single base are about 17 to about 121 nucleotides in length, preferably about 17 to about 74 nucleotides in length. Most preferably, however, the oligonucleotides of the present invention are at least about 25 bases in length, unless there are self-dimerization structures within the oligonucleotide. If the oligonucleotide has such an unfavorable structure, lengths longer than 35 bases are preferred. Oligonucleotides with modified ends both shorter and longer than certain of the exemplified, modified oligonucleotides herein function as gene repair or gene knockout agents and are within the scope of the present invention.

Once an oligomer is chosen, it can be tested for its tendency to self-dimerize, since self-dimerization may result in reduced efficiency of alteration of genetic information. Checking for self-dimerization tendency can be accomplished manually or, preferably, using a software program. One such program is Oligo Analyzer 2.0, available through Integrated DNA Technologies (Coralville, Iowa 52241) (http://www.idtdna.com); this program is available for use on the world wide web at http://www.idtdna.com/program/oligoanalyzer/oligoanalyzer.asp.

For each oligonucleotide sequence input into the program, Oligo Analyzer 2.0 reports possible self-dimerized duplex forms, which are usually only partially duplexed, along with the free energy change associated with such self-dimerization. Delta G-values that are negative and large in magnitude, indicating strong self-dimerization potential, are automatically flagged by the software as “bad”. Another software program that analyzes oligomers for pair dimer formation is Primer Select from DNASTAR, Inc., 1228 S. Park St., Madison, Wis. 53715, Phone: (608) 258-7420 (http://www.dnastar.com/products/PrimerSelect.html).

If the sequence is subject to significant self-dimerization, the addition of further sequence flanking the “repair” nucleotide can improve gene correction frequency.

Generally, the oligonucleotides of the present invention are identical in sequence to one strand of the target DNA, which can be either strand of the target DNA, with the exception of one or more targeted bases positioned within the DNA domain of the oligonucleotide, and preferably toward the middle between the modified terminal regions. Preferably, the difference in sequence of the oligonucleotide as compared to the targeted genomic DNA is located at about the middle of the oligo-nucleotide sequence. In a preferred embodiment, the oligonucleotides of the invention are complementary to the non-transcribed strand of a duplex. In other words, the preferred oligonucleotides target the sense strand of the DNA, i.e. the oligonucleotides of the invention are preferably complementary to the strand of the target DNA the sequence of which is found in the mRNA.

The oligonucleotides of the invention can include more than a single base change. In an oligonucleotide that is about a 70-mer, with at least one modified residue incorporated on the ends, as disclosed herein, multiple bases can be simultaneously targeted for change. The target bases may be up to 27 nucleotides apart and may not be changed together in all resultant plasmids in all cases. There is a frequency distribution such that the closer the target bases are to each other in the central DNA domain within the oligonucleotides of the invention, the higher the frequency of change in a given cell. Target bases only two nucleotides apart are changed together in every case that has been analyzed. The farther apart the two target bases are, the less frequent the simultaneous change. Thus, oligonucleotides of the invention may be used to repair or alter multiple bases rather than just one single base. For example, in a 74-mer oligonucleotide having a central base targeted for change, a base change event up to about 27 nucleotides away can also be effected. The positions of the altering bases within the oligonucleotide can be optimized using any one of the assays described herein. Preferably, the altering bases are at least about 8 nucleotides from one end of the oligonucleotide.

The oligonucleotides of the present invention can be introduced into cells by any suitable means. According to certain preferred embodiments, the modified oligonucleotides may be used alone. Suitable means, however, include the use of polycations, cationic lipids, liposomes, polyethylenimine (PEI), electroporation, biolistics, microinjection and other methods known in the art to facilitate cellular uptake. For plant cells, biolistic or particle bombardment methods are typically used. According to certain preferred embodiments of the present invention, isolated plant cells are treated in culture according to the methods of the invention, to mutate or repair a target gene. Alternatively, plant target DNA may be modified in vitro or in another cell type, including for example, yeast or bacterial cells and then introduced into a plant cell as, for example, a T-DNA. Plant cells thus modified may be used to regenerate the whole organism as, for example, in a plant having a desired targeted genomic change. In other instances, targeted genomic alteration, including repair or mutagenesis, may take place in vivo following direct administration of the modified, single-stranded oligonucleotides of the invention to a subject.

The single-stranded, modified oligonucleotides of the present invention have numerous applications as gene repair, gene modification, or gene knockout agents. Such oligonucleotides may be advantageously used, for example, to introduce or correct multiple point mutations. Each mutation leads to the addition, deletion or substitution of at least one base pair. The methods of the present invention offer distinct advantages over other methods of altering the genetic makeup of an organism, in that only the individually targeted bases are altered. No additional foreign DNA sequences are added to the genetic complement of the organism. Such agents may, for example, be used to develop plants with improved traits by rationally changing the sequence of selected genes in isolated cells and using these modified cells to regenerate whole plants having the altered gene. See, e.g., U.S. Pat. No. 6,046,380 and U.S. Pat. No. 5,905,185 incorporated herein by reference. Such plants produced using the compositions of the invention lack additional undesirable selectable markers or other foreign DNA sequences. Targeted base pair substitution or frameshift mutations introduced by an oligonucleotide in the presence of a cell-free extract also provides a way to modify the sequence of extrachromosomal elements, including, for example, plasmids, cosmids and artificial chromosomes. The oligonucleotides of the invention also simplify the production of plants having particular modified or inactivated genes. Altered plant model systems such as those produced using the methods and oligonucleotides of the invention are invaluable in determining the function of a gene and in evaluating drugs. The oligonucleotides and methods of the present invention may also be used to introduce molecular markers, including, for example, SNPs, RFLPs, AFLPs and CAPs.

The purified oligonucleotide compositions may be formulated in accordance with routine procedures depending on the target. For example, purified oligonucleotide can be used directly in a standard reaction mixture to introduce alterations into targeted DNA in vitro or where cells are the target as a composition adapted for bathing cells in culture or for microinjection into cells in culture. The purified oligonucleotide compositions may also be provided on coated microbeads for biolistic delivery into plant cells. Where necessary, the composition may also include a solubilizing agent. Generally, the ingredients will be supplied either separately or mixed together in single-use form, for example, as a dry, lyophilized powder or water-free concentrate. In general, dosage required for efficient targeted gene alteration will range from about 0.001 to 50,000 μg/kg target tissue, preferably between 1 to 250 μg/kg, and most preferably at a concentration of between 30 and 60 micromolar.

For cell administration, direct injection into the nucleus, biolistic bombardment, electroporation, liposome transfer and calcium phosphate precipitation may be used. In yeast, lithium acetate or spheroplast transformation may also be used. In a preferred method, the administration is performed with a liposomal transfer compound, e.g., DOTAP (Boehringer-Mannheim) or an equivalent such as lipofectin. The amount of the oligonucleotide used is about 500 nanograms in 3 micrograms of DOTAP per 100,000 cells. For electroporation, between 20 and 2000 nanograms of oligonucleotide per million cells to be electroporated is an appropriate range of dosages which can be increased to improve efficiency of genetic alteration upon review of the appropriate sequence according to the methods described herein. For biolistic delivery, microbeads are generally coated with resuspended oligonucleotides, which range of oligonucleotide to microbead concentration can be similarly adjusted to improve efficiency as determined using one of the assay methods described herein, starting with about 0.05 to 1 microgram of oligonucleotide to 25 microgram of 1.0 micrometer gold beads or similar microcarrier.

Another aspect of the invention is a kit comprising at least one oligonucleotide of the invention. The kit may comprise an additional reagent or article of manufacture. The additional reagent or article of manufacture may comprise a delivery mechanism, cell extract, a cell, or a plasmid, such as one of those disclosed in the Figures herein, for use in an assay of the invention. Alternatively, the invention includes a kit comprising an isogenic set of cells in which each cell in the kit comprises a different altered amino acid for a target protein encoded by a targeted altered gene within the cell produced according to the methods of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Flow diagram for the generation of modified single-stranded oligonucleotides. The upper strands of chimeric oligonucleotides I and II are separated into pathways resulting in the generation of single-stranded oligonucleotides that contain (A) 2′-O-methyl RNA nucleotides or (B) phosphorothioate linkages. Fold changes in repair activity for correction of kan[0040] ^sin the HUH7 cell-free extract are presented in parenthesis. HUH7 cells are described in Nakabayashi et al., Cancer Research 42: 3858-3863 (1982). Each single-stranded oligonucleotide is 25 bases in length and contains a G residue mismatched to the complementary sequence of the kan^sgene. The numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2′-O-methyl RNA nucleotides (R) are at each end of the molecule. Hence oligo 12S/25G contains an all phosphorothioate backbone, displayed as a dotted line. Smooth lines indicate DNA residues, wavy lines indicate 2′-O-methyl RNA residues and the carat indicates the mismatched base site (G). FIG. 1(C) provides a schematic plasmid indicating the sequence of the kan chimeric double-stranded hairpin oligonucleotide (left; SEQ ID NO: 2673) and the sequence the tet chimeric double-stranded hairpin oligonucleotide used in other experiments (right; SEQ ID NO: 2674). FIG. 1(D) provides a flow chart of a kan experiment in which a chimeric double-stranded hairpin oligonucleotide (SEQ ID NO: 2673) is used. In FIG. 1(D), the Kan mutant sequence corresponds to SEQ ID NO: 2675 and SEQ ID NO: 2676; the Kan converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678; the mutant sequence in the sequence trace corresponds to SEQ ID NO: 2679 and the converted sequences in the sequence trace correspond to SEQ ID NO: 2680.
FIG. 2. Genetic readout system for correction of a point mutation in plasmid pK[0041] ^sm4021. A mutant kanamycin gene harbored in plasmid pK^sm4021 is the target for correction by oligonucleotides. The mutant G is converted to a C by the action of the oligo. Corrected plasmids confer resistance to kanamycin in E.coli (DH10B) after electroporation leading to the genetic readout and colony counts. The wild type sequence corresponds to SEQ ID NO: 2681.
FIG. 3: Target plasmid and sequence correction of a frameshift mutation by chimeric and single-stranded oligonucleotides. (A) Plasmid pT[0042] ^sΔ208 contains a single base deletion mutation at position 208 rendering it unable to confer tet resistance. The target sequence presented below indicates the insertion of a T directed by the oligonucleotides to re-establish the resistant phenotype. (B) DNA sequence confirming base insertion directed by Tet 3S/25G; the yellow highlight indicates the position of frameshift repair. The wild type sequence corresponds to SEQ ID NO: 2682, the mutant sequence corresponds to SEQ ID NO: 2683 and the converted sequence corresponds to SEQ ID NO: 2684. The control sequence in the sequence trace corresponds to SEQ ID NO: 2685 and the 3S/25A sequence in the sequence trace corresponds to SEQ ID NO: 2686.
FIG. 4. DNA sequences of representative kan[0043] ^rcolonies. Confirmation of sequence alteration directed by the indicated molecule is presented along with a table outlining codon distribution. Note that 10S/25G and 12S/25G elicit both mixed and unfaithful gene repair. The number of clones sequenced is listed in parentheses next to the designation for the single-stranded oligonucleotide. A plus (+) symbol indicates the codon identified while a figure after the (+) symbol indicates the number of colonies with a particular sequence. TAC/TAG indicates a mixed peak. Representative DNA sequences are presented below the table with yellow highlighting altered residues. The sequences in the sequence traces have been assigned numbers as follows: 3S/25G, 6S/25G and 8S/25G correspond to SEQ ID NO: 2687, 10S/25G corresponds to SEQ ID NO: 2688, 25S/25G on the lower left corresponds to SEQ ID NO: 2689 and 25S/25G on the lower right corresponds to SEQ ID NO: 2690.
FIG. 5. Gene correction in HeLa cells. Representative oligonucleotides of the invention are co-transfected with the pCMVneo([0044] ⁻)FIAsH plasmid (shown in FIG. 9) into HeLa cells. Ligand is diffused into cells after co-transfection of plasmid and oligonucleotides. Green fluorescence indicates gene correction of the mutation in the antibiotic resistance gene. Correction of the mutation results in the expression of a fusion protein that carries a marker ligand binding site and when the fusion protein binds the ligand, a green fluorescence is emitted. The ligand is produced by Aurora Biosciences and can readily diffuse into cells enabling a measurement of corrected protein function; the protein must bind the ligand directly to induce fluorescence. Hence cells bearing the corrected plasmid gene appear green while “uncorrected” cells remain colorless.
FIG. 6. Z-series imaging of corrected cells. Serial cross-sections of the HeLa cell represented in FIG. 5 are produced by Zeiss 510 LSM confocal microscope revealing that the fusion protein is contained within the cell. [0045]
FIG. 7. Hygromycin-eGFP target plasmids. (A) Plasmid pAURHYG(ins)GFP contains a single base insertion mutation between nucleotides 136 and 137, at codon 46, of the Hygromycin B coding sequence (cds) which is transcribed from the constitutive ADH1 promoter. The target sequence presented below indicates the deletion of an A and the substitution of a C for a T directed by the oligonucleotides to re-establish the resistant phenotype. In FIG. 7A, the sequence of the normal allele corresponds to SEQ ID NO: 2691, the sequence of the targe/existing mutation corresponds to SEQ ID NO: 2692 and the sequence of the desired alteration corresponds to SEQ ID NO: 2693. (B) Plasmid pAURHYG(rep)GFP contains a base substitution mutation introducing a G at nucleotide 137, at codon 46, of the Hygromycin B coding sequence (cds). The target sequence presented below the diagram indicates the amino acid conservative replacement of G with C, restoring gene function. In FIG. 7B, the sequence of the normal allele correspond to SEQ ID NO: 2691, the sequence of the targe/existing mutation corresponds to SEQ ID NO: 2694 and the sequence of the desired alteration corresponds to SEQ ID NO: 2693. [0046]
FIG. 8. Oligonucleotides for correction of hygromycin resistance gene. The sequence of the oligonucleotides used in experiments to assay correction of a hygromycin resistance gene are shown. DNA residues are shown in capital letters, RNA residues are shown in lowercase and nucleotides with a phosphorothioate backbone are capitalized and underlined. In FIG. 8, the sequence of HygE3T/25 corresponds to SEQ ID NO: 2695, the sequence of HygE3T/74 corresponds to SEQ ID NO: 2696, the sequence of HygE3T/74a corresponds to SEQ ID NO: 2697, the sequence of HygGG/Rev corresponds to SEQ ID NO: 2698 and the sequence of Kan70T corresponds to SEQ ID NO: 2699. [0047]
FIG. 9. pAURNeo(−)FIAsH plasmid. This figure describes the plasmid structure, target sequence, oligonucleotides, and the basis for detection of the gene alteration event by fluorescence. In FIG. 9, the sequence of the Neo/kan target mutant corresponds to SEQ ID NO: 2675 and SEQ ID NO: 2676, the converted sequence corresponds to SEQ ID NO: 2677 and SEQ ID NO: 2678 and the FIAsH peptide sequence corresponds to SEQ ID NO: 2700. [0048]
FIG. 10. pYESHyg(x)eGFP plasmid. This plasmid is a construct similar to the pAURHyg(x)eGFP construct shown in FIG. 7, except the promoter is the inducible GAL1 promoter. This promoter is inducible with galactose, leaky in the presence of raffinose, and repressed in the presence of dextrose. [0049]
FIG. 11. pBI-HygeGFP plasmid. This plasmid is a construct based on the plasmids pBI101, pBI 101.2, pBI101.3 or pBI 121 available from Clontech in which HygeGFP replaces the beta-glucuronidase gene of the Clontech plasmids. The different Clontech plasmids vary by a reading frame shift relative to the polylinker, or the presence of the Cauliflower mosaic virus promoter.[0050]
The following examples are provided by way of illustration only, and are not intended to limit the scope of the invention disclosed herein. [0051]

EXAMPLE 1

Assay Method for Base Alteration and Preferred Oligonucleotide Selection

In this example, single-stranded and double-hairpin oligonucleotides with chimeric backbones (see FIG. 1 for structures (A and B) and sequences (C and D) of assay oligonucleotides) are used to correct a point mutation in the kanamycin gene of pK[0052] ^sm4021 (FIG. 2) or the tetracycline gene of pT^sΔ208 (FIG. 3). All kan oligonucleotides share the same 25 base sequence surrounding the target base identified for change, just as all tet oligonucleotides do. The sequence is given in FIG. 1C and FIG. 1D. Each plasmid contains a functional ampicillin gene. Kanamycin gene function is restored when a G at position 4021 is converted to a C (via a substitution mutation); tetracycline gene function is restored when a deletion at position 208 is replaced by a C (via frameshift mutation). A separate plasmid, pAURNeo(−)FIAsH (FIG. 9), bearing the kan^sgene is used in the cell culture experiments. This plasmid was constructed by inserting a synthetic expression cassette containing a neomycin phosphotransferasea (kanamycin resistance) gene and an extended reading frame that encodes a receptor for the FIAsH ligand into the pAUR123 shuttle vector (Panvera Corp., Madison, Wis.). The resulting construct replicates in S. cerevisiae at low copy number, confers resistance to aureobasidinA and constitutively expresses either the Neo+/FIAsH fusion product (after alteration) or the truncated Neo−/FIAsH product (before alteration) from the ADH1 promoter. By extending the reading frame of this gene to code for a unique peptide sequence capable of binding a small ligand to form a fluorescent complex, restoration of expression by correction of the stop codon can be detected in real time using confocal microscopy.
Additional constructs can be made to test additional gene alteration events or for specific use in different expression systems. For example, alternative comparable plant plasmids or integration vectors such as, e.g. those based on T-DNA, can be constructed for stable expression in plant cells according to the disclosures herein. Such constructs would use a plant specific promoter such as, e.g., cauliflower mosaic virus 35S promoter, to replace the promoters directing expression of the neo, hyg or aureobasidinA resistance gene disclosed herein, including for example, in FIGS. 7B, 9 and [0053] 10 herein. Moreover, the green fluorescent protein (GFP) sequence used herein may be modified to increase expression in plant cells such as Arabidopsis and the other plants disclosed herein as described in Haseloff et al., Proc. Natl.Acad. Sci. 94(6): 2122-7 (1997), Rouwendal et al. Plant Mol. Biol. 33(6): 989-99 (1997) and Hu et al. FEBS Lett. 369(2-3): 331-4 (1995). Codon usage for optimal expression of GFP in plants results from increasing the frequency of codons with a C or a G in the third position from 32 to about 60%. Specific constructs are disclosed and can be used as follows with such plant specific alterations.
We also construct three mammalian expression vectors, pHyg(rep)eGFP, pHyg(Δ)eGFP, pHyg(ins)eGFP, that contain a substitution mutation at nucleotide 137 of the hygromycin-B coding sequence. (rep) indicates a T1374→G replacement, (Δ) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. All point mutations create a nonsense termination codon at residue 46. We use pHYGeGFP plasmid (Invitrogen, CA) DNA as a template to introduce the mutations into the hygromycin-eGFP fusion gene by a two step site-directed mutagenesis PCR protocol. First, we generate overlapping 5′ and a 3′ amplicons surrounding the mutation site by PCR for each of the point mutation sites. A 215 [0054] bp 5′ amplicon for the (rep), (Δ) or (ins) was generated by polymerization from oligonucleotide primer HygEGFPf (5′-AATACGACTCACTATAGG-3′; SEQ ID NO: 2701) to primer Hygrepr (5′GACCTATCCACGCCCTCC-3′; SEQ ID NO: 2702), HygΔr (5′-GACTATCCACGCCCTCC-3′; SEQ ID NO: 2703), or Hyginsr (5′-GACATTATCCACGCCCTCC-3′; SEQ ID NO: 2704), respectively. We generate a 300 bp 3′ amplicon for the (rep), (Δ) or (ins) by polymerization from oligonucleotide primers Hygrepf (5′-CTGGGATAGGTCCTGCGG-3′; SEQ ID NO: 2705), HygΔf (5′-CGTGGATAGTCCTGCGG-3′; SEQ ID NO: 2706), Hyginsf (5′-CGTGGATAATGTCCTGCGG-3′; SEQ ID NO: 2707), respectively to primer HygEGFPr (5′-AAATCACGCCATGTAGTG-3′; SEQ ID NO: 2708). We mix 20 ng of each of the resultant 5′ and 3′ overlapping amplicon mutation sets and use the mixture as a template to amplify a 523 bp fragment of the Hygromycin gene spanning the KpnI and RsrII restriction endonuclease sites. We use the Expand PCR system (Roche) to generate all amplicons with 25 cycles of denaturing at 94° C. for 10 seconds, annealing at 55° C. for 20 seconds and elongation at 68° C. for 1 minute. We digest 10 μg of vector pHYGeGFP and 5 μg of the resulting fragments for each mutation with KpnI and RsrII (NEB) and gel purify the fragment for enzymatic ligation. We ligate each mutated insert into pHYGeGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm the mutation by Sanger dideoxy chain termination sequencing and purify the plasmid using a Qiagen maxiprep kit.
Oligonucleotide synthesis and cells. Chimeric oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) are synthesized using available phosphoramidites on controlled pore glass supports. After deprotection and detachment from the solid support, each oligonucleotide is gel-purified using, for example, procedures such as those described in Gamper et al., [0055] Biochem. 39, 5808-5816 (2000) and the concentrations determined spectrophotometrically (33 or 40 μg/ml per A₂₆₀unit of single-stranded or hairpin oligomer). HUH7 cells are grown in DMEM, 10% FBS, 2 mM glutamine, 0.5% pen/strep. The E.coli strain, DH10B, is obtained from Life Technologies (Gaithersburg, Md.); DH10B cells contain a mutation in the RECA gene (recA).
Cell-free extracts. Although this portion of this example is directed to mammalian systems, similar extracts from plants can be prepared as disclosed elsewhere in this application and used as disclosed in this example. We prepare cell-free extracts from HUH7 cells or other mammalian cells, as follows. We employ this protocol with essentially any mammalian cell including, for example, H1299 cells (human epithelial carcinoma, non-small cell lung cancer), C127I (immortal murine mammary epithelial cells), MEF (mouse embryonic fibroblasts), HEC-1-A (human uterine carcinoma), HCT15 (human colon cancer), HCT116 (human colon carcinoma), LoVo (human colon adenocarcinoma), and HeLa (human cervical carcinoma). We harvest approximately 2×10[0056] ⁸cells. We then wash the cells immediately in cold hypotonic buffer (20 mM HEPES, pH7.5; 5 mM KCl; 1.5 mM MgCl₂; 1 mM DTT) with 250 mM sucrose. We then resuspend the cells in cold hypotonic buffer without sucrose and after 15 minutes we lyse the cells with 25 strokes of a Dounce homogenizer using a tight fitting pestle. We incubate the lysed cells for 60 minutes on ice and centrifuge the sample for 15 minutes at 12000×g. The cytoplasmic fraction is enriched with nuclear proteins due to the extended co-incubation of the fractions following cell breakage. We then immediately aliquote and freeze the supernatant at −80° C. We determine the protein concentration in the extract by the Bradford assay.
We also perform these experiments with cell-free extracts obtained from fungal cells, including, for example, [0057] S. cerevisiae (yeast), Ustilago maydis, and Candida albicans. For example, we grow yeast cells into log phase in 2L YPD medium for 3 days at 30° C. We then centrifuge the cultures at 5000×g, resuspend the pellets in a 10% sucrose, 50 mM Tris, 1 mM EDTA lysis solution and freeze them on dry ice. After thawing, we add KCl, spermidine and lyticase to final concentrations of 0.25 mM, 5 mM and 0.1 mg/ml, respectively. We incubate the suspension on ice for 60 minutes, add PMSF and Triton X100 to final concentrations of 0.1 mM and 0.1% and continue to incubate on ice for 20 minutes. We centrifuge the lysate at 3000×g for 10 minutes to remove larger debris. We then remove the supernatant and clarify it by centrifuging at 30000×g for 15 minutes. We then add glycerol to the clarified extract to a concentration of 10% (v/v) and freeze aliquots at −80° C. We determine the protein concentration of the extract by the Bradford assay.
Reaction mixtures of 50 μl are used, consisting of 10-30 μg protein of cell-free extract, which can be optionally substituted with purified proteins or enriched fractions, about 1.5 μg chimeric double-hairpin oligonucleotide or 0.55 μg single-stranded molecule (3S/25G or 6S/25G, see FIG. 1), and 1 μg of plasmid DNA (see FIGS. 2 and 3) in a reaction buffer of 20 mM Tris, pH 7.4, 15 mM MgCl[0058] ₂, 0.4 mM DTT, and 1.0 mM ATP. Reactions are initiated with extract and incubated at 30° C. for 45 min. The reaction is stopped by placing the tubes on ice and then immediately deproteinized by two phenol/chloroform (1:1) extractions. Samples are then ethanol precipitated. The nucleic acid is pelleted at 15,000 r.p.m. at 4° C. for 30 min., is washed with 70% ethanol, resuspended in 50 μl H₂O, and is stored at −20° C. 5 μl of plasmid from the resuspension (˜100 ng) was transfected in 20 μl of DH10B cells by electroporation (400 V, 300 μF, 4 kΩ) in a Cell-Porator apparatus (Life Technologies). After electroporation, cells are transferred to a 14 ml Falcon snap-cap tube with 2 ml SOC and shaken at 37° C. for 1 h. Enhancement of final kan colony counts is achieved by then adding 3 ml SOC with 10 μg/ml kanamycin and the cell suspension is shaken for a further 2 h at 37° C. Cells are then spun down at 3750×g and the pellet is resuspended in 500 μl SOC. 200 μl is added undiluted to each of two kanamycin (50 μg/ml) agar plates and 200 μl of a 10⁵dilution is added to an ampicillin (100 μg/ml) plate. After overnight 37° C. incubation, bacterial colonies are counted using an Accucount 1000 (Biologics). Gene conversion effectiveness is measured as the ratio of the average of the kan colonies on both plates per amp colonies multiplied by 10⁻⁵to correct for the amp dilution.
The following procedure can also be used. 5 μl of resuspended reaction mixtures (total volume 50 μl) are used to transform 20 μl aliquots of electro-competent DH10B bacteria using a Cell-Porator apparatus (Life Technologies). The mixtures are allowed to recover in 1 ml SOC at 37° C. for 1 hour at which time 50 μg/ml kanamycin or 12 μg/ml tetracycline is added for an additional 3 hours. Prior to plating, the bacteria are pelleted and resuspended in 200 μl of SOC. 100 μl aliquots are plated onto kan or tet agar plates and 100 μl of a 10[0059] ^{31 4}dilution of the cultures are concurrently plated on agar plates containing 100 μg/ml of ampicillin. Plating is performed in triplicate using sterile Pyrex beads. Colony counts are determined by an Accu-count 1000 plate reader (Biologics). Each plate contains 200-500 ampicillin resistant colonies or 0-500 tetracycline or kanamycin resistant colonies. Resistant colonies are selected for plasmid extraction and DNA sequencing using an ABI Prism kit on an ABI 310 capillary sequencer (PE Biosystems).
Chimeric single-stranded oligonucleotides. In FIG. 1 the upper strands of chimeric oligonucleotides I and II are separated into pathways resulting in the generation of single-stranded oligo-nucleotides that contain (FIG. 1A) 2′-O-methyl RNA nucleotides or (FIG. 1B) phosphorothioate linkages. Fold changes in repair activity for correction of kan[0060] ^sin the HUH7 cell-free extract are presented in parenthesis. Each single-stranded oligonucleotide is 25 bases in length and contains a G residue mismatched to the complementary sequence of the kan^sgene.
Molecules bearing 3, 6, 8, 10 and 12 phosphorothioate linkages in the terminal regions at each end of a backbone with a total of 24 linkages (25 bases) are tested in the kan[0061] ^ssystem. Alternatively, molecules bearing 2, 4, 5, 7, 9 and 11 in the terminal regions at each end are tested. The results of one such experiment, presented in Table 1 and FIG. 1B, illustrate an enhancement of correction activity directed by some of these modified structures. In this illustrative example, the most efficient molecules contained 3 or 6 phosphorothioate linkages at each end of the 25-mer; the activities are approximately equal (molecules IX and X with results of 3.09 and 3.7 respectively). A reduction in alteration activity may be observed as the number of modified linkages in the molecule is further increased. Interestingly, a single-strand molecule containing 24 phosphorothioate linkages is minimally active suggesting that this backbone modification when used throughout the molecule supports only a low level of targeted gene repair or alteration. Such a non-altering, completely modified molecule can provide a baseline control for determining efficiency of correction for a specific oligonucleotide molecule of known sequence in defining the optimum oligonucleotide for a particular alteration event.
The efficiency of gene repair directed by phosphorothioate-modified, single-stranded molecules, in a length dependent fashion, led us to examine the length of the RNA modification used in the original chimera as it relates to correction. Construct III represents the “RNA-containing” strand of chimera I and, as shown in Table 1 and FIG. 2A, it promotes inefficient gene repair. But, as shown in the same figure, reducing the RNA residues on each end from 10 to 3 increases the frequency of repair. At equal levels of modification, however, 25-mers with 2′-O-methyl ribonucleotides were less effective gene repair agents than the same oligomers with phosphorothioate linkages. These results reinforce the fact that an RNA containing oligonucleotide is not as effective in promoting gene repair or alteration as a modified DNA oligonucleotide. [0062]
Repair of the kanamycin mutation requires a G→C exchange. To confirm that the specific desired correction alteration was obtained, colonies selected at random from multiple experiments are processed and the isolated plasmid DNA is sequenced. As seen in FIG. 4, colonies generated through the action of the single-stranded [0063] molecules 3S/25G (IX), 6S/25G (X) and 8S/25G (XI) respectively contained plasmid molecules harboring the targeted base correction. While a few colonies appeared on plates derived from reaction mixtures containing 25-mers with 10 or 12 thioate linkages on both ends, the sequences of the plasmid molecules from these colonies contain nonspecific base changes. In these illustrative examples, the second base of the codon is changed (see FIG. 3). These results show that modified single-strands can direct gene repair, but that efficiency and specificity are reduced when the 25-mers contain 10 or more phosphorothioate linkages at each end.
In FIG. 1, the [0064] numbers 3, 6, 8, 10, 12 and 12.5 respectively indicate how many phosphorothioate linkages (S) or 2′-O-methyl RNA nucleotides (R) are at each end of the examplified molecule although other molecules with 2, 4, 5, 7, 9 and 11 modifications at each end can also be tested. Hence oligo 12S/25G represents a 25-mer oligonucleotide which contains 12 phosphorothioate linkages on each side of the central G target mismatch base producing a fully phosphorothioate linked backbone, displayed as a dotted line. The dots are merely representative of a linkage in the figure and do not depict the actual number of linkages of the oligonucleotide. Smooth lines indicate DNA residues, wavy lines indicate 2′-O-methyl RNA residues and the carat indicates the mismatched base site (G).
Correction of a mutant kanamycin gene in cultured mammalian cells. Although this portion of this example is directed to cultured mammalian cells, comparable methods may be used using cultured plant cells or protoplasts of those cells from the plant species disclosed herein. The experiments are performed using different eukaryotic cells including plant and mammalian cells, including, for example, 293 cells (transformed human primary kidney cells), HeLa cells (human cervical carcinoma), and H1299 (human epithelial carcinoma, non-small cell lung cancer). HeLa cells are grown at 37° C. and 5% CO[0065] ₂in a humidified incubator to a density of 2×10⁵cells/ml in an 8 chamber slide (Lab-Tek). After replacing the regular DMEM with Optimem, the cells are co-transfected with 10 μg of plasmid pAURNeo(−) FIAsH and 5 μg of modified single-stranded oligonucleotide (3S/25G) that is previously complexed with 10 μg lipofectamine, according to the manufacturer's directions (Life Technologies). The cells are treated with the liposome-DNA-oligo mix for 6 hrs at 37° C. Treated cells are washed with PBS and fresh DMEM is added. After a 16-18 hr recovery period, the culture is assayed for gene repair. The same oligonucleotide used in the cell-free extract experiments is used to target transfected plasmid bearing the kan^sgene. Correction of the point mutation in this gene eliminates a stop codon and restores full expression. This expression can be detected by adding a small non-fluorescent ligand that bound to a C-C-R-E-C-C sequence (SEQ ID NO: 2717) in the genetically modified carboxy terminus of the kan protein, to produce a highly fluorescent complex (FIAsH system, Aurora Biosciences Corporation). Following a 60 min incubation at room temperature with the ligand (FIAsH-EDT2), cells expressing full length kan product acquire an intense green fluorescence detectable by fluorescence microscopy using a fluorescein filter set. Similar experiments are performed using the HygeGFP target as described in Example 2 with a variety of mammalian cells, including, for example, COS-1 and COS-7 cells (African green monkey), and CHO-K1 cells (Chinese hamster ovary). The experiments are also performed with PG12 cells (rat pheochromocytoma) and ES cells (human embryonic stem cells).
Summary of experimental results. Tables 1, 2 and 3 respectively provide data on the efficiency of gene repair directed by single-stranded oligonucleotides. Table 1 presents data using a cell-free extract from human liver cells (HUH7) to catalyze repair of the point mutation in plasmid pkan[0066] ^sm4021 (see FIG. 1). Table 2 illustrates that the oligomers are not dependent on MSH2 or MSH3 for optimal gene repair activity. Table 3 illustrates data from the repair of a frameshift mutation (FIG. 3) in the tet gene contained in plasmid pTetΔ208. Table 4 illustrates data from repair of the pkan^sm4021 point mutation catalyzed by plant cell extracts prepared from canola and musa (banana). Colony numbers are presented as kan^ror tet^rand fold increases (single strand versus double hairpin) are presented for kan^rin Table 1.
FIG. 5A is a confocal picture of HeLa cells expressing the corrected fusion protein from an episomal target. Gene repair is accomplished by the action of a modified single-stranded oligonucleotide containing 3 phosphorothioate linkages at each end (3S/25G). FIG. 5B represents a “Z-series” of HeLa cells bearing the corrected fusion gene. This series sections the cells from bottom to top and illustrates that the fluorescent signal is “inside the cells”. [0067]
Results. In summary, we have designed a novel class of single-stranded oligonucleotides with backbone modifications at the termini and demonstrate gene repair/conversion activity in mammalian and plant cell-free extracts. We confirm that the all DNA strand of the RNA-DNA double-stranded double hairpin chimera is the active component in the process of gene repair. In some cases, the relative frequency of repair by the novel oligonucleotides of the invention is elevated approximately 3-4-fold in certain embodiments when compared to frequencies directed by chimeric RNA-DNA double hairpin oligonucleotides. [0068]
This strategy centers around the use of extracts from various sources to correct a mutation in a plasmid using a modified single-stranded or a chimeric RNA-DNA double hairpin oligonucleotide. A mutation is placed inside the coding region of a gene conferring antibiotic resistance in bacteria, here kanamycin or tetracycline. The appearance of resistance is measured by genetic readout in [0069] E.coli grown in the presence of the specified antibiotic. The importance of this system is that both phenotypic alteration and genetic inheritance can be measured. Plasmid pK^sm4021 contains a mutation (T→G) at residue 4021 rendering it unable to confer antibiotic resistance in E.coli. This point mutation is targeted for repair by oligonucleotides designed to restore kanamycin resistance. To avoid concerns of plasmid contamination skewing the colony counts, the directed correction is from G→C rather than G→T (wild-type). After isolation, the plasmid is electroporated into the DH10B strain of E.coli, which contains inactive RecA protein. The number of kanamycin colonies is counted and normalized by ascertaining the number of ampicillin colonies, a process that controls for the influence of electroporation. The number of colonies generated from three to five independent reactions was averaged and is presented for each experiment. A fold increase number is recorded to aid in comparison.
The original RNA-DNA double hairpin chimera design, e.g., as disclosed in U.S. Pat. No. 5,565,350, consists of two hybridized regions of a single-stranded oligonucleotide folded into a double hairpin configuration. The double-stranded targeting region is made up of a 5 base pair DNA/DNA segment bracketed by 10 base pair RNA/DNA segments. The central base pair is mismatched to the corresponding base pair in the target gene. When a molecule of this design is used to correct the kan[0070] ^smutation, gene repair is observed (I in FIG. 1A). Chimera II (FIG. 1B) differs partly from chimera I in that only the DNA strand of the double hairpin is mismatched to the target sequence. When this chimera was used to correct the kan^smutation, it was twice as active. In the same study, repair function could be further increased by making the targeting region of the chimera a continuous RNA/DNA hybrid.
Frame shift mutations are repaired. By using plasmid pT[0071] ^sΔ208, described in FIG. 1(C) and FIG. 3, the capacity of the modified single-stranded molecules that showed activity in correcting a point mutation, can be tested for repair of a frameshift. To determine efficiency of correction of the mutation, a chimeric oligonucleotide (Tet I), which is designed to insert a T residue at position 208, is used. A modified single-stranded oligonucleotide (Tet IX) directs the insertion of a T residue at this same site. FIG. 3 illustrates the plasmid and target bases designated for change in the experiments. When all reaction components are present (extract, plasmid, oligomer), tetracycline resistant colonies appear. The colony count increases with the amount of oligonucleotide used up to a point beyond which the count falls off (Table 3). No colonies above background are observed in the absence of either extract or oligonucleotide, nor when a modified single-stranded molecule bearing perfect complementarity is used. FIG. 3 represents the sequence surrounding the target site and shows that a T residue is inserted at the correct site. We have isolated plasmids from fifteen colonies obtained in three independent experiments and each analyzed sequence revealed the same precise nucleotide insertion. These data suggest that the single-stranded molecules used initially for point mutation correction can also repair nucleotide deletions.
Comparison of phosphorothioate oligonucleotides to 2′-O-methyl substituted oligonucleotides. From a comparison of molecules VII and XI, it is apparent that gene repair is more subject to inhibition by RNA residues than by phosphorothioate linkages. Thus, even though both of these oligonucleotides contain an equal number of modifications to impart nuclease resistance, XI (with 16 phosphorothioate linkages) has good gene repair activity while VII (with 16 2′-O-methyl RNA residues) is inactive. Hence, the original chimeric double hairpin oligonucleotide enabled correction directed, in large part, by the strand containing a large region of contiguous DNA residues. [0072]
Oligonucleotides can target multiple nucleotide alterations within the same template. The ability of individual single-stranded oligonucleotides to correct multiple mutations in a single target template is tested using the plasmid pK[0073] ^sm4021 and the following single-stranded oligonucleotides modified with 3 phosphorothioate linkages at each end (indicated as underlined nucleotides): Oligo1 is a 25-mer with the sequence TTCGATAAGCCTATGCTGACCCGTG (SEQ ID NO: 2709) corrects the original mutation present in the kanamycin resistance gene of pK^sm4021 as well as directing another alteration 2 basepairs away in the target sequence (both indicated in boldface); Oligo2 is a 70-mer with the 5′-end sequence TTCGGCTACGACTGGGCACAACAGACAATTGGC (SEQ ID NO: 2710) with the remaining nucleotides being completely complementary to the kanamycin resistance gene and also ending in 3 phosphorothioate linkages at the 3′ end. Oigo2 directs correction of the mutation in pK^sm4021 as well as directing another alteration 21 basepairs away in the target sequence (both indicated in boldface).
We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pK[0074] ^sM4021 plasmid. These include, for example, a second 25-mer that alters two nucleotides that are three nucleotides apart with the sequence 5′-TTGTGCCCAGTCGTATCCGAATAGC-3′ (SEQ ID NO: 2711); a 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5′-CATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTAGCCGAATAGCCTCTCCACCCAAGCGGCCGGAGA-3′ (SEQ ID NO: 2712); and another 70-mer that alters two nucleotides that are 21 nucleotides apart with the sequence 5′-GCTGACAGCCGGAACACGGCGGCATCAGAGCAGCCAATTGTCTGTTGTGCCCAGTCGTAGCCGMTAGCCT-3′ (SEQ ID NO: 2713). The nucleotides in the oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are modified in the same way as the other oligonucleotides of the invention.
We assay correction of the original mutation in pK[0075] ^sm4021 by monitoring kanamycin resistance (the second alterations which are directed by Oligo2 and Oligo3 are silent with respect to the kanamycin resistance phenotype). In addition, in experiments with Oligo2, we also monitor cleavage of the resulting plasmids using the restriction enzyme Tsp5091 which cuts at a specific site present only when the second alteration has occurred (at ATT in Oligo2). We then sequence these clones to determine whether the additional, silent alteration has also been introduced. The results of an analysis are presented below:

Oligo 1 (25-mer) Oligo 2 (70-mer)

Clones with both sites changed 9 7

Clones with a single site changed 0 2

Clones that were not changed 4 1
Nuclease sensitivity of unmodified DNA oligonucleotide. Electrophoretic analysis of nucleic acid recovered from the cell-free extract reactions conducted here confirm that the unmodified single-stranded 25-mer did not survive incubation whereas greater than 90% of the terminally modified oligos did survive (as judged by photo-image analyses of agarose gels). [0076]
Plant extracts direct repair. The modified single-stranded constructs can be tested in plant cell extracts. We have observed gene alteration using extracts from multiple plant sources, including, for example, Arabidopsis, tobacco, banana, maize, soybean, canola, wheat, spinach as well as spinach chloroplast extract or extracts made from other plant cells disclosed herein. We prepare the extracts by grinding plant tissue or cultured cells under liquid nitrogen with a mortar and pestle. We extract 3 ml of the ground plant tissue with 1.5 ml of extraction buffer (20 mM HEPES, pH7.5; 5 mM KCl; 1.5 mM MgCl[0077] ₂; 10 mM DTT; and 10% [v/v] glycerol). Some plant cell-free extracts also include about 1% (w/v) PVP. We then homogenize the samples with 15 strokes of a Dounce homogenizer. Following homogenization, we incubate the samples on ice for 1 hour and centrifuge at 3000×g for 5 minutes to remove plant cell debris. We then determine the protein concentration in the supernatants (extracts) by Bradford assay. We dispense 100 μg (protein) aliquots of the extracts which we freeze in a dry ice-ethanol bath and store at −80° C.
We describe experiments using two sources here: a dicot (canola) and a monocot (banana, [0078] Musa acuminata cv. Rasthali). Each vector directs gene repair of the kanamycin mutation (Table 4); however, the level of correction is elevated 2-3 fold relative to the frequency observed with the chimeric oligonucleotide. These results are similar to those observed in the mammalian system wherein a significant improvement in gene repair occurred when modified single-stranded molecules were used.

Tables are attached hereto.

TABLE I


Gene repair activity is directed by single-stranded oligonucleotides.

Oligonucleotide	Plasmid	Extract (ug)	kan^rcolonies	Fold increase

I	pK^Sm4021	10	300
I	↓	20	418	1.0 ×
II	↓	10	537
II	↓	20	748	1.78 ×
III	↓	10	3
III	↓	20	5	0.01 ×
IV	↓	10	112
IV	↓	20	96	0.22 ×
V	↓	10	217
V	↓	20	342	0.81 ×
VI	↓	10	6
VI	↓	20	39	0.093 ×
VII	↓	10	0
VII	↓	20	0	0 ×
VIII	↓	10	3
VIII	↓	20	5	0.01 ×
IX	↓	10	936
IX	↓	20	1295	3.09 ×
X	↓	10	1140
X	↓	20	1588	3.7 ×
XI	↓	10	480
XI	↓	20	681	1.6 ×
XII	↓	10	18
XII	↓	20	25	0.059 ×
XIII	↓	10	0
XIII	↓	20	4	0.009 ×
—	↓	20	0
I	↓	—	0

Plasmid pK ^Sm4021 (1 μg), the indicated oligonucleotide (1.5 μg chimeric oligonucleotide or 0.55 μg single-stranded oligonucleotide; molar ratio of oligo to plasmid of 360 to 1) and either 10 or 20 μg of HUH7 cell-free extract were incubated 45 min at 37° C. Isolated plasmid DNA was electroporated into E. coli (strain DH10B) and the number of kan^rcolonies counted. The data represent the number of kanamycin resistant colonies per 10⁶ampicillin resistant colonies generated from the same reaction and is the average of three experiments (standard deviation usually less than +/−15%). Fold increase is defined relative to 418 kan^rcolonies (second reaction) and in all reactions was calculated using the 20 μg sample.

TABLE II


Modified single-stranded oligomers are not dependent on MSH2
or MSH3 for optimal gene repair activity.

	A.	Oligonucleotide	Plasmid	Extract	kan^rcolonies

IX (3S/25G)	↓	HUH7	637
X (6S/25G)	↓	HUH7	836
IX	↓	MEF2^−/−	781
X	↓	MEF2^−/−	676
IX	↓	MEF3^−/−	582
X	↓	MEF3^−/−	530
IX	↓	MEF^+/+	332
X	↓	MEF^+/+	497
—	↓	MEF2^−/−	10
—	↓	MEF3 ^−/−	5
—	↓	MEF^+/+	14

Chimeric oligonucleotide (1.5 μg) or modified single-stranded oligonucleotide (0.55 μg) was incubated with 1 μg of plasmid pK ^Sm4021 and 20 μg of the indicated extracts. MEF represents mouse embryonic fibroblasts with either MSH2 (2^−/−) or MSH3 (3^−/−) deleted. MEF^+/+ indicates wild-type mouse embryonic fibroblasts. The other reaction components were then added and processed through the bacterial readout system. The data represent the number of kanamycin resistant colonies per 10⁶ampicillin resistant colonies.

TABLE III


Frameshift mutation repair is directed by
single-stranded oligonucleotides

Oligonucleotide	Plasmid	Extract	tet^rcolonies

Tet IX (3S/25A; 0.5 μg)	pT^SΔ208 (1 μg)		—	0
—	↓	20 μg		0
Tet IX (0.5 μg)	↓	↓		48
Tet IX (1.5 μg)	↓	↓		130
Tet IX (2.0 μg)	↓	↓		68
Tet I (chimera; 1.5 μg)	↓	↓		48

Each reaction mixture contained the indicated amounts of plasmid and oligonucleotide. The extract used for these experiments came from HUH7 cells. The data represent the number of tetracycline resistant colonies per 10 ⁶ampicillin resistant colonies generated from the same reaction and is the average of 3 independent experiments. Tet I is a chimeric oligonucleotide and Tet IX is a modified single-stranded oligonucleotide that are designed to insert a T residue at position 208 of pT^sΔ208. The oligonucleotides are equivalent to structures I and IX in FIG. 2.

TABLE IV


Plant cell-free extracts support gene repair by
single-stranded oligonucleotides

Oligonucleotide	Plasmid	Extract	kan^rcolonies

II (chimera)	pK^Sm402l	30 μg	Canola	337
IX (3S/25G)	↓		Canola	763
X (6S/25G)	↓		Canola	882
II	↓		Musa	203
IX	↓		Musa	343
X	↓		Musa	746
—	↓		Canola	0
—	↓		Musa	0
IX	↓	—	Canola	0
X	↓	—	Musa	0

Canola or Musa cell-free extracts were tested for gene repair activity on the kanamycin-sensitive gene as previously described in (18). Chimeric oligonucleotide II (1.5 μg) and modified single-stranded oligonucleotides IX and X (0.55 μg) were used to correct pK[0083] ^Sm4021. Total number of kan^rcolonies are present per 10⁷ampicillin resistant colonies and represent an average of four independent experiments.

TABLE V

Gene repair activity in cell-free extracts prepared from yeast

(Saccharomyces cerevisiae)

Cell-type Plasmid Chimeric Oligo SS Oligo kan^r/amp^r× 10⁶

Wild type pKan^sm4021 1 μg 0.36

Wild type ↓ 1 μg 0.81

ΔRAD52 ↓ 1 μg 10.72

ΔRAD52 ↓ 1 μg 17.41

ΔPMS1 ↓ 1 μg 2.02

ΔPMS1 ↓ 1 μg 3.23

EXAMPLE 2

Yeast Cell Targeting Assay Method for Base Alteration and Preferred Oligonucleotide Selection

In this example, single-stranded oligonucleotides with modified backbones and double-hairpin oligonucleotides with chimeric, RNA-DNA backbones are used to measure gene repair using two episomal targets with a fusion between a hygromycin resistance gene and eGFP as a target for gene repair. These plasmids are pAURHYG(rep)GFP, which contains a point mutation in the hygromycin resistance gene (FIG. 7), pAURHYG(ins)GFP, which contains a single-base insertion in the hygromycin resistance gene (FIG. 7) and pAURHYG(Δ)GFP which has a single base deletion. We also use the plasmid containing a wild-type copy of the hygromycin-eGFP fusion gene, designated pAURHYG(wt)GFP, as a control. These plasmids also contain an aureobasidinA resistance gene. In pAURHYG(rep)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when a G at position 137, at codon 46 of the hygromycin B coding sequence, is converted to a C thus removing a premature stop codon in the hygromycin resistance gene coding region. In pAURHYG(ins)GFP, hygromycin resistance gene function and green fluorescence from the eGFP protein are restored when an A inserted between nucleotide positions 136 and 137, at codon 46 of the hygromycin B coding sequence, is deleted and a C is substituted for the T at position 137, thus correcting a frameshift mutation and restoring the reading frame of the hygromycin-eGFP fusion gene. [0084]
We synthesize the set of three yeast expression constructs pAURHYG(rep)eGFP, pAURHYG(Δ)eGFP, pAURHYG(ins)eGFP, that contain a point mutation at nucleotide 137 of the hygromycin-B coding sequence as follows. (rep) indicates a T137→G replacement, (Δ) represents a deletion of the G137 and (ins) represents an A insertion between nucleotides 136 and 137. We construct this set of plasmids by excising the respective expression cassettes by restriction digest from pHyg(x)EGFP and ligation into pAUR123 (Panvera, Calif.). We digest 10 μg pAUR123 vector DNA, as well as, 10 μg of each pHyg(x)EGFP construct with KpnI and SaII (NEB). We gel purify each of the DNA fragments and prepare them for enzymatic ligation. We ligate each mutated insert into pHygEGFP vector at 3:1 molar ratio using T4 DNA ligase (Roche). We screen clones by restriction digest, confirm by Sanger dideoxy chain termination sequencing and purify using a Qiagen maxiprep kit. [0085]
We use this system to assay the ability of five oligonucleotides (shown in FIG. 8) to support correction under a variety of conditions. The oligonucleotides which direct correction of the mutation in pAURHYG(rep)GFP can also direct correction of the mutation in pAURHYG(ins)GFP. Three of the four oligonucleotides (HygE3T/25, HygE3T/74 and HygGG/Rev) share the same 25-base sequence surrounding the base targeted for alteration. HygGG/Rev is an RNA-DNA chimeric double hairpin oligonucleotide of the type described in the prior art. One of these oligonucleotides, HygE3T/74, is a 74-base oligonucleotide with the 25-base sequence centrally positioned. The fourth oligonucleotide, designated HygE3T/74α, is the reverse complement of HygE3T/74. The fifth oligonucleotide, designated Kan70T, is a non-specific, control oligonucleotide which is not complementary to the target sequence. Alternatively, an oligonucleotide of identical sequence but lacking a mismatch to the target or a completely thioate modified oligonucleotide or a completely 2-O-methylated modified oligonucleotide may be used as a control. Alternatively, oligonucleotides containing one, two, three, four, five, six, eight, ten or more LNA modifications on at least one of the two termini (and preferrably the 3′ terminus) may be used in different embodiments. [0086]
Oligonucleotide synthesis and cells. We synthesized and purified the chimeric, double-hairpin oligonucleotides and single-stranded oligonucleotides (including those with the indicated modifications) as described in Example 1. Plasmids used for assay were maintained stably in yeast ([0087] Saccharomyces cerevisiae) strain LSY678 MAT α at low copy number under aureobasidin selection. Plasmids and oligonucleotides are introduced into yeast cells by electroporation as follows: to prepare electrocompetent yeast cells, we inoculate 10 ml of YPD media from a single colony and grow the cultures overnight with shaking at 300 rpm at 30° C. We then add 30 ml of fresh YPD media to the overnight cultures and continue shaking at 30° C. until the OD₆₀₀was between 0.5 and 1.0 (3-5 hours). We then wash the cells by centrifuging at 4° C. at 3000 rpm for 5 minutes and twice resuspending the cells in 25 ml ice-cold distilled water. We then centrifuge at 4° C. at 3000 rpm for 5 minutes and resuspend in 1 ml ice-cold 1M sorbitol and then finally centrifuge the cells at 4° C. at 5000 rpm for 5 minutes and resuspend the cells in 120 μl 1M sorbitol. To transform electrocompetent cells with plasmids or oligonucleotides, we mix 40 μl of cells with 5 μg of nucleic acid, unless otherwise stated, and incubate on ice for 5 minutes. We then transfer the mixture to a 0.2 cm electroporation cuvette and electroporate with a BIO-RAD Gene Pulser apparatus at 1.5 kV, 25 μF, 200 Ω for one five-second pulse. We then immediately resuspend the cells in 1 ml YPD supplemented with 1M sorbitol and incubate the cultures at 30° C. with shaking at 300 rpm for 6 hours. We then spread 200 μl of this culture on selective plates containing 300 μg/ml hygromycin and spread 200 μl of a 10⁵dilution of this culture on selective plates containing 500 ng/ml aureobasidinA and/or and incubate at 30° C. for 3 days to allow individual yeast colonies to grow. We then count the colonies on the plates and calculate the gene conversion efficiency by determining the number of hygromycin resistance colonies per 10⁵aureobasidinA resistant colonies.
Frameshift mutations are repaired in yeast cells. We test the ability of the oligonucleotides shown in FIG. 8 to correct a frameshift mutation in vivo using LSY678 yeast cells containing the plasmid pAURHYG(ins)GFP. These experiments, presented in Table 6, indicate that these oligonucleotides can support gene correction in yeast cells. These data reinforce the results described in Example 1 indicating that oligonucleotides comprising phosphorothioate linkages facilitate gene correction much more efficiently than control duplex, chimeric RNA-DNA oligonucleotides. This gene correction activity is also specific as transformation of cells with the control oligonucleotide Kan70T produced no hygromycin resistant colonies above background and thus Kan70T did not support gene correction in this system. In addition, we observe that the 74-base oligonucleotide (HygE3T/74) corrects the mutation in pAURHYG(ins)GFP approximately five-fold more efficiently than the 25-base oligonucleotide (HygE3T/25). We also perform control experiments with LSY678 yeast cells containing the plasmid pAURHYG(wt)GFP. With this strain we observed that even without added oligonucleotides, there are too many hygromycin resistant colonies to count. [0088]
We also use additional oligonucleotides to assay the ability of individual oligonucleotides to correct multiple mutations in the pAURHYG(x)eGFP plasmid. These include, for example, one that alters two basepairs that are 3 nucleotides apart is a 74-mer with the [0089] sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGGTACGTCCTGCGGGTAAATAGCTGCGCCGATGGTTTCTAC-3′ (SEQ ID NO: 2714); a 74-mer that alters two basepairs that are 15 nucleotides apart with the sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTAAACAGCTGCGCCGATGGTTTCTAC-3′ (SEQ ID NO: 2715); and a 74-mer that alters two basepairs that are 27 nucleotides apart with the sequence 5′-CTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATACGTCCTGCGGGTAAATAGCTGCGCCGACGGTTTCTAC (SEQ ID NO: 2716). The nucleotides in these oligonucleotides that direct alteration of the target sequence are underlined and in boldface. These oligonucleotides are modified in the same ways as the other oligonucleotides of the invention.
Oligonucleotides targeting the sense strand direct gene correction more efficiently. We compare the ability of single-stranded oligonucleotides to target each of the two strands of the target sequence of both pAURHYG(ins)GFP and pAURHYG(rep)GFP. These experiments, presented in Tables 7 and 8, indicate that an oligonucleotide, HygE3T/74α, with sequence complementary to the sense strand (i.e. the strand of the target sequence that is identical to the mRNA) of the target sequence facilitates gene correction approximately ten-fold more efficiently than an oligonucleotide, HygE3T/74, with sequence complementary to the non-transcribed strand which serves as the template for the synthesis of RNA. As indicated in Table 7, this effect was observed over a range of oligonucleotide concentrations from 0-3.6 μg, although we did observe some variability in the difference between the two oligonucleotides (indicated in Table 7 as a fold difference between HygE3T/74α and HygE3T/74). Furthermore, as shown in Table 8, we observe increased efficiency of correction by HygE3T/74α relative to HygE3T/74 regardless of whether the oligonucleotides were used to correct the base substitution mutation in pAURHYG(rep)GFP or the insertion mutation in pAURHYG(ins)GFP. The data presented in Table 8 further indicate that the single-stranded oligonucleotides correct a base substitution mutation more efficiently than an insertion mutation. However, this last effect was much less pronounced and the oligonucleotides of the invention are clearly able efficiently to correct both types of mutations in yeast cells. In addition, the role of transcription is investigated using plasmids with inducible promoters such as that described in FIG. 10. [0090]
Optimization of oligonucleotide concentration. To determine the optimal concentration of oligonucleotide for the purpose of gene alteration, we test the ability of increasing concentrations of Hyg3T/74α to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678. We chose this assay system because our previous experiments indicated that it supports the highest level of correction. However, this same approach could be used to determine the optimal concentration of any given oligonucleotide. We test the ability of Hyg3T/74α to correct the mutation in pAURHYG(rep)GFP contained in yeast LSY678 over a range of oligonucleotide concentrations from 0-10.0 μg. As shown in Table 9, we observe that the correction efficiency initially increases with increasing oligonucleotide concentration, but then declines at the highest concentration tested. [0091]
Tables are attached hereto. [0092]

TABLE 6

Correction of an insertion mutation in pAURHYG(ins)GFP by

HygGG/Rev, HygE3T/25 and HygE3T/74

Colonies on Colonies on Correction

Oligonucleotide Tested Hygromycin Aureobasidin (/10⁵) Efficiency

HygGG/Rev 3 157 0.02

HygE3T/25 64 147 0.44

HygE3T/74 280 174 1.61

Kan70T 0 — —
[0093]

TABLE 7

An oligonucleotide targeting the sense strand of the target sequence

corrects more efficiently.

Colonies per

hygromycin plate

Amount of Oligonucleotide (μg) HygE3T/74 HygE3T/74α

0 0 0

0.6 24 128 (8.4x)*

1.2 69 140 (7.5x)*

2.4 62 167 (3.8x)*

3.6 29 367 (15x)*
[0094]

TABLE 8

Correction of a base substitution mutation is more efficient than correction

of a frame shift mutation.

Oligonucleotide Plasmid tested (contained in LSY678)

Tested (5 μg) pAURHYG(ins)GFP pAURHYG(rep)GFP

HygE3T/74 72 277

HygE3T/74α 1464 2248

Kan70T 0 0
[0095]

TABLE 9

Optimization of oligonucleotide concentration in electroporated yeast cells.

Colonies on Colonies on Correction

Amount (μg) hygromycin aureobasidin (/10⁵) efficiency

0 0 67 0

1.0 5 64 0.08

2.5 47 30 1.57

5.0 199 33 6.08

7.5 383 39 9.79

10.0 191 33 5.79

EXAMPLE 3

Cultured Cell Manipulation

Although disclosure in this example is directed to use of stem cells or human blood cells and microinjection, the microinjection procedures may also be used with cultured plant cells or protoplasts using any plant species, including those disclosed herein. Mononuclear cells are isolated from human umbilical cord blood of normal donors using Ficoll Hypaque (Pharmacia Biotech, Uppsala, Sweden) density centrifugation. CD34+ cells are immunomagnetically purified from mononuclear cells using either the progenitor or Multisort Kits (Miltenyi Biotec, Auburn, Calif.). Lin[0096] ⁻CD38⁻ cells are purified from the mononuclear cells using negative selection with StemSep system according to the manufacturer's protocol (Stem Cell Technologies, Vancouver, Calif.). Cells used for microinjection are either freshly isolated or cryopreserved and cultured in Stem Medium (S Medium) for 2 to 5 days prior to microinjection. S Medium contains Iscoves' Modified Dulbecc's Medium without phenol red (IMDM) with 100 μg/ml glutamine/penicillin/streptomycin, 50 mg/ml bovine serum albumin, 50 μg/ml bovine pancreatic insulin, 1 mg/ml human transferrin, and IMDM; Stem Cell Technologies), 40 μg/ml low-density lipoprotein (LDL; Sigma, St. Louis, Mo.), 50 mM HEPEs buffer and 50 μM 2-mercaptoethanol, 20 ng/ml each of thrombopoietin, flt-3 ligand, stem cell factor and human IL-6 (Pepro Tech Inc., Rocky Hill, N.J.). After microinjection, cells are detached and transferred in bulk into wells of 48 well plates for culturing.
35 mm dishes are coated overnight at 4° C. with 50 μg/ml Fibronectin (FN) fragment CH-296 (Retronectin; TaKaRa Biomedicals, Panvera, Madison, Wis.) in phosphate buffered saline and washed with IMDM containing glutamine/penicillin/streptomycin. 300 to 2000 cells are added to cloning rings and attached to the plates for 45 minutes at 37° C. prior to microinjection. After incubation, cloning rings are removed and 2 ml of S Medium are added to each dish for microinjection. Pulled injection needles with a range of 0.22 μm to 0.3 μm outer tip diameter are used. Cells are visualized with a microscope equipped with a temperature controlled stage set at 37° C. and injected using an electronically interfaced Eppendorf Micromanipulator and Transjector. Successfully injected cells are intact, alive and remain attached to the plate post injection. Molecules that are flourescently labeled allow determination of the amount of oligonucleotide delivered to the cells. [0097]
For in vitro erythropoiesis from Lin[0098] ⁻CD38⁻ cells, the procedure of Malik, 1998 can be used. Cells are cultured in ME Medium for 4 days and then cultured in E Medium for 3 weeks. Erythropoiesis is evident by glycophorin A expression as well as the presence of red color representing the presence of hemoglobin in the cultured cells. The injected cells are able to retain their proliferative capacity and the ability to generate myeloid and erythoid progeny. CD34+ cells can convert a normal A (β^A) to sickle T (β^S) mutation in the β-globin gene or can be altered using any of the oligonucleotides of the invention herein for correction or alteration of a normal gene to a mutant gene. Alternatively, stem cells can be isolated from blood of humans having genetic disease mutations and the oligonucleotides of the invention can be used to correct a defect or to modify genomes within those cells.
Alternatively, non-stem cell populations of cultured cells can be manipulated using any method known to those of skill in the art including, for example, the use of polycations, cationic lipids, liposomes, polyethylenimine (PEI), electroporation, biolistics, calcium phosphate precipitation, or any other method known in the art. [0099]
Biolistic delivery of oligonucleotide into plant cells may be accomplished according to the following method. One milliliter of packed cell volume of plant cell suspensions are subcultured onto plates containing solid medium [with Murashige and Skoog salts from Gibco/BRL, 500 mg/liter Mes, 1 mg/liter thiamin, 100 mg/liter myo-inositol, 180 mg/liter KH2PO4, 2.21 mg/[0100] liter 2,4-dichlorophenoxyacetic acid (2,4-D), and 30 g/liter sucrose (pH 5.7) and having 8 g/liter agar-agar from Sigma added before autoclaving]. By using a helium-driven particle gun such as that from BioRad and following manufacturers directions, oligonucleotides may be introduced to cells after precipitation onto 1 micrometer or comparable gold microcarriers (Bio-Rad). To precipitate onto microcarriers, 35 microliters of a particle suspension (60 mg of microcarriers per ml of 100% ethanol) is transferred to a 1.5 ml microcentrifuge tube, which is agitated on a vortex mixer. Then 40 microliter of resuspended oligonucleotide (60 ng/microliter water) is added; then 75 microliter of ice-cold 2.5 M CaCl2 is added; then 75 microliter of ice-cold 0.1 M spermidine is added. The tube is mixed vigorously or a vortex mixer for 10 min at room temperature. The particles are allowed to settle for 10 min and are centrifuged at 11,750 g for 30 sec. The supernatant is removed and the particles are resuspended in 50 microliter of 100% ethanol. An aliquot of 10 microliter of the resuspended particles are applied to each macro-projectile which is used to bombard each plate once at 900 psi (1 psi=6.89 kPa) with a gap distance (distance from power source to macroprojectile) of 1 cm and a target distance (distance from microprojectile launch site to target material) of 10 cm.
An alternative method of delivery can be used as follows. Cultured cells are suspended in liquid N6 medium and then plated on a VWR Scientific glass fiber filter. About 0.4 microgram of oligonucleotide are precipitated with 15 microliter of 2.5 mM CaCl2 and 5 microliter of 0.1 M spermidine onto 25 microgram of 1.0 micrometer gold particles. Microprojectile bombardment is performed by using a Bio-Rad PDS-1000 He particle delivery system or comparable machine following manufacturers instructions. Alterations in oligonucleotide concentrations can be employed to determine the optimum concentration of oligonucleotide according to the procedures described herein for any particular oligonucleotide of the invention. [0101]
Alternatively, the oligonucleotide of the invention may be delivered to a plant cell by electroporation of a protoplast derived from a plant part. The protoplasts may be formed by enzymatic treatment of a plant part, particularly a leaf, according to techniques such as those in Gallois et al., Methods in Molecular Biology 55: 89-107 by Humana Press. Such conditions for electroporation use about 3×10[0102] ⁵protoplasts in a total volume of about 0.3 ml with a concentration of oligonucleotide of between 0.6 to 4 microgram per ml.

EXAMPLE 4

Plant Cells

The oligonucleotides of the invention can also be used to repair or direct a mutagenic event in plants and animal cells. Although little information is available on plant mutations amongst natural cultivars, the oligonucleotides of the invention can be used to produce “knock out” mutations by modification of specific amino acid codons to produce stop codons (e.g., a CAA codon specifying Gln can be modified at a specific site to TAA; a AAG codon specifying Lys can be modified to UAG at a specific site; and a CGA codon for Arg can be modified to a UGA codon at a specific site). Such base pair changes will terminate the reading frame and produce a defective truncated protein, shortened at the site of the stop codon. [0103]
Alternatively, frameshift additions or deletions can be directed into the genome at a specific sequence to interrupt the reading frame and produce a garbled downstream protein. Such stop or frameshift mutations can be introduced to determine the effect of knocking out the protein in either plant or animal cells. [0104]
For introduction of a T-DNA, including the T-DNA in the plasmid of FIG. 11, into a plant cell, [0105] Agrobacterium tumefaciens is used. These techniques are routine standard techniques known in the art. For example, one method follows. We transform A. tumefaciens is transformed by electroporation (using a BioRad Gene Pulser™). Competent A. tumefaciens is prepared using a method similar to that of preparing competent E. coli by suspending a freshly grown culture three times in ice-cold water and a final resuspension in 10% glycerol. Electroporation conditions are a 0.2 cm gap cuvette at a setting of 25 μF,200 Ω and2.5 kV.
[0106] A. tumefaciens containing a plasmid with a T-DNA is then used to introduce the T-DNA into a plant cell using routine standard techniques known in the art. For example, we transform Arabidopsis by vacuum infiltration or by dipping flowers in an Agrobacterium solution containing a surfactant, e.g. L-77. Seeds are then collected, grown and screened for presence of the T-DNA. Alternatively, Agrobacterium can be used to transform callus tissue and the callus tissue can then be used to regenerate transformed plants.
All publications and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. [0107]
Notes on the Tables Presented Below: [0108]
Each of the following tables presents, for the specified gene, a plurality of mutations that are known to confer a relevant phenotype and, for each mutation, the oligonucleotides that can be used to correct the respective mutation site-specifically in the genome according to the present invention. [0109]
The left-most column identifies each alteration or mutation and the phenotype that the alteration/mutation confers. [0110]
For most entries, the mutation/alteration is identified at both the nucleic acid and protein level. At the amino acid level, mutations are presented according to the following standard nomenclature. The centered number identifies the position of the mutated codon in the protein sequence; to the left of the number is the wild type residue and to the right of the number is the mutant codon. Terminator codons are shown as “TERM”. At the nucleic acid level, the entire triplet of the wild type and mutated codons is shown. [0111]
The middle column presents, for each mutation, four oligonucleotides capable of repairing the mutation site-specifically in the genome or in cloned DNA including DNA in artificial chromosomes, episomes, plasmids, or other types of vectors. The oligonucleotides of the invention, however, may include any of the oligonucleotides sharing portions of the sequence of the 121 base sequence. Thus, oligonucleotides of the invention for each of the depicted targets may be 18, 19, 20 up to about 121 nucleotides in length. Sequence may be added non-symmetrically. [0112]
All oligonucleotides are presented, per convention, in the 5′ to 3′ orientation. The nucleotide that effects the change in the genome is underlined and presented in bold. [0113]
The first of the four oligonucleotides for each mutation is a 121 nt oligonucleotide centered about the repair/altering nucleotide. The second oligonucleotide, its reverse complement, targets the opposite strand of the DNA duplex for repair/alteration. The third oligonucleotide is the minimal 17 nt domain of the first oligonucleotide, also centered about the repair/alteration nucleotide. The fourth oligonucleotide is the reverse complement of the third, and thus represents the minimal 17 nt domain of the second. [0114]
The third column of each table presents the SEQ ID NO: of the respective repair oligonucleotide. [0115]

EXAMPLE 5

Engineering Herbicide Resistant Plants

Chemical weed control is an important tool of modern agriculture and many herbicides have been developed for this purpose. Their use has resulted in substantial increases in the yields of many crops, including, for example, maize, soybeans, and cotton. Thus while the use of fertilizers and new high-yielding crop varieties have contributed greatly to the “green revolution,” chemical weed control has also been at the forefront of technological achievement. [0116]
Herbicides having broad-spectrum activity are particularly useful because they obviate the need for multiple herbicides targeting different classes of weeds. The problem with such herbicides is that they typically also affect crops which are exposed to the herbicide. One way to overcome this is to generate plants which are resistant to one or more broad-spectrum herbicides. Such herbicide-tolerant plants may reduce the need for tillage to control weeds, thereby effectively reducing soil erosion and can reduce the quantity and number of different herbicides applied in the field. [0117]
Common herbicides used, for example, include those that inhibit the enzyme 5-enolpyruvyl-3-phosphoshikimic acid synthase (EPSPS), for example N-phosphonomethyl-glycine (e.g. glyphosate), those that inhibit acetolactate synthase (ALS) activity, for example the sulfonylureas and related herbicides, and those that inhibit dihydropteroate synthase, for example methyl[(4-amino-phenyl)sulfonyl]carbamate (e.g. Asulam). Herbicide-tolerant plants can be produced by several methods, including, for example, introducing into the genome of the plant the ability to degrade the herbicide, the capacity to produce a higher level of the targeted enzyme, and/or expressing an herbicide-tolerant allele of the enzyme. [0118]

The attached tables disclose exemplary oligonucleotides base sequences which can be used to generate site-specific mutations in plant genes that confer herbicide resistance.

TABLE 10


Genome-Altering Oligos Conferring Glyphosate Resistance

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Glyphosate Resistance	AAGCGTCGGAGATTGTACTTCAACCCATTTAGAGAAATCTCCGGTC	1
EPSPS	TTATTAAGCTTCCTGCCTCCAAGTCTCTATCAAATCGGATCCTGC
Arabidopsis thaliana	TTCTCGCTGCTCTGTCTGAGGTATATATCAC
Gly97Ala	GTGATATATACCTCAGACAGAGCAGCGAGAAGCAGGATCCGATT	2
GGC-GCC	TGATAGAGACTTGGAGGCAGGAAGCTTAATAAGACCGGAGATTT
	CTCTAATGGGTTGAAGTACAATCTCCGACGCTT
	GCTTCCTG C CTCCAAGT	3
	ACTTGGAG G CAGGAAGC	4

Glyphosate Resistance	AAGCTTCAGAGATTGTGCTTCAACCAATCAGAGAAATCTCGGGTC	5
EPSPS	TCATTAAGCTACCCGCATCCAAATCTCTCTCCAATCGGATCCTCC
Brassica napus	TTCTTGCCGCTCTATCTGAGGTACATATACT
Gly93AIa	AGTATATGTACCTCAGATAGAGCGGCAAGAAGGAGGATCCGATT	6
GGA-GCA	GGAGAGAGATTTGGATGCGGGTAGCTTAATGAGACCCGAGATTT
	CTCTGATTGGTTGAAGCACAATCTCTGAAGCTT
	GCTACCCG C ATCCAAAT	7
	ATTIGGAT G CGGGTAGC	8

Glyphosate Resistance	AGCCCAACGAGATTGTGCTGCAACCCATCAAAGATATATCAGGC	9
EPSPS 1	ACTGTTAAATTGCCTGCTTCTAAATCCCTTTCCAATCGTATTCTCC
Nicotiana tabacum	TTCTTGCTGCCCTTTCTAAGGGAAGGACTGT
Gly95Ala	ACAGTCCTTCCCTTAGAAAGGGCAGCAAGAAGGAGAATACGATT	10
GGT-GCT	GGAAAGGGATTTAGAA G CAGGCAATTTAACAGTGCCTGATATATC
	TTTGATGGGTTGCAGCACAATCTCGTIGGGCT
	ATTGCCTG C TTCTAAAT	11
	ATTTAGAA G CAGGCAAT	12

Glyphosate Resistance	ATTGTTTCCTTGGTACGAAATGTCCTCCTGTTCGAATTGTCAGCA	13
EPSPS 2	AGGGAGGCCTTCCCGCAGGGAAGGTAAAGCTCTCTGGATCAATT
Nicotiana tabacum	AGCAGCCAGTACTTGACTGCTCTGCTTATGGC
Gly62Ala	GCCATAAGCAGAGCAGTCAAGTACTGGCTGCTAATTGATCCAGA	14
GGA-GCA	GAGCTTTACCTTCCCT G CGGGAAGGCCTCCCTTGCTGACAATTC
	GAACAGGAGGACATTTCGTACCAAGGAAACAAT
	CCTTCCCG C AGGGAAGG	15
	CCTTCCCG C GGGAAGG	16

Glyphosate Resistance	ATTGTTTCCTTGGCACTGACTGGCCACCTGTTCGTGTCAATGGAA	17
EPSPS	TCGGAGGGCTACCTG C TGGCAAGGTCAAGCTGTCTGGCTCCATC
Zea mays	AGCAGTCAGTACTTGAGTGCCTTGCTGATGGC
Gly168Ala	GCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAGCCAGA	18
GGT-GCT	CAGCTTGACCTTGCCA G CAGGTAGCCCTCCGATTCCATTGACAC
	GAACAGGTGGGCAGTCAGTGCCAAGGAAACAAT
	GCTACCTG C TGGCAAGG	19
	CCTTGCCA G CAGGTAGC	20

Glyphosate Resistance	ACTGTTTCCTTGGCACTGAATGCCCACCTGTTCGTGTCAAGGGA	21
EPSPS	ATTGGAGGACTTCCTG C TGGCAAGGTTAAGCTCTCTGGTTCCAT
Cryza sativa	CAGCAGTCAGTACTTGAGTGCCTTGCTGATGGC
Gly115Ala	GCCATCAGCAAGGCACTCAAGTACTGACTGCTGATGGAACCAGA	22
GGT-GCT	GAGCTTAACCTTGCCAGCAGGAAGTCCTCCAATTCCCTTGACAC
	GAACAGGTGGGCATTCAGTGCCAAGGAAACAGT
	ACTTCCTG C TGGCAAGG	23
	CCTTGCCA G CAGGAAGT	24

Glyphosate Resistance	AGCCTTCTGAGATAGTGTTGCAACCCATTAAAGAGATTTCAGGCA	25
EPSPS	CTGTTAAATTGCCTGCCTCTAAATCATTATCTAATAGAATTCTCCT
Petunia x hybrida	TCTTGCTGCCTTATCTGAAGGMCAACTGT
Gly93Ala	ACAGTTGTTCCTTCAGATAAGGCAGCAAGAAGGAGAATTCTATTA	26
GGC-GCC	GATAATGATTTAGAGGCAGGCAATTTAACAGTGCCTGAAATCTCT
	TTAATGGGTTGCAACACTATCTCAGAAGGCT
	ATTGCCTG CCTCTAAAT	27
	ATTTAGAG G CAGGCAAT	28

Glyphosate Resistance	AACCCCATGAGATTGTGCTAGNACCCATCAAAGATATATCTGGTA	29
EPSPS	CTGTTAAATTACCCG C TTCGAAATCCCTTTCCAATCGTATTCTCCT
Lycopersicon	TCTTGCTGCCCTTTCTGAGGGAAGGACTGT
esculentum	ACAGTCCTTCCCTCAGAAAGGGCAGCAAGAAGGAGAATACGATT	30
Gly97Ala	GGAAAGGGATTTCGAA G CGGGTAATTTAACAGTACCAGATATATC
GGT-GCT	TTTGATGGGTNCTAGCACAATCTGATGGGGTT
	ATTACCCG C TTCGAAAT	31
	ATTTCGAA G CGGGTAAT	32

Glyphosate Resistance	ATTGTTTCCTTGGCACTGACTGCCCACCTGTTCGKATCAACGGGA	33
EPSPS	TTGGAGGGCTACCTGCTGGCAAGGTTAAGCTGTCTGGTTCCAIT
Lolium rigidum	AGCAGCCAATACTTGAGTTCCTTGCTGATGGC
Gly107Ala	GCCATCAGCAAGGAACTCAAGTATTGGCTGCTGATGGAACCAGA	34
GGT-GCT	CAGCTTAACCTTGCCA G CAGGTAGCCCTCCAATGCCGTTGATCG
	AACAGGTGGGCAGTCAGTGCCAAGGAAACAAT
	GCTACCTG C TGGCAAGG	35
	CCTTGCCA G CAGGTAGC	36

TABLE 11


Genome-Altering Oligos Conferring Imidazolinone
and Sulfonylurea Herbicide Resistance

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Sulfonylurea	AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA	37
Resistance	ATCACAGGACAAGTC T CTCGTCGTATGATTGGTACAGATGCGTTT
ALS	CAAGAGACTCCGATTGTTGAGGTAACGCGTT
Arabidopsis thaliana	AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC	38
Pro197Ser	CAATCATACGACGAG A GACTTGTCCTGTGATTGCTACAAGAGGAA
CCT-TCT	CACTATCTAACAACGCATCGGCTAATCCGCT
	GACAAGTCTC T CGTCGT	39
	ACGACGAG A GACTTGTC	40

Sulfonylurea	AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA	41
Resistance	ATCACAGGACAAGTCC AG CGTCGTATGATTGGTACAGATGCGTTT
ALS	CAAGAGACTCCGATTGTTGAGGTAACGCGTT
Arabidopsis thaliana	AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC	42
Pro197GLN	CAATCATACGACG C TGGACTTGTCCTGTGATTGCTACAAGAGGAA
CCT-CAG	CACTATCTAACAACGCATCGGCTAATCCGCT
	ACAAGTCC AG CGTCGTC	43
	TACGACG CT GGACTTGT	44

Sulfonylurea	AGCGGATTAGCCGATGCGTTGTTAGATAGTGTTCCTCTTGTAGCA	45
Resistance	ATCACAGGACAAGTCC AA CGTCGTATGATTGGTACAGATGCGTTT
ALS	CAAGAGACTCCGATTGTTGAGGTAACGCGTT
Arabidopsis thaliana	AACGCGTTACCTCAACAATCGGAGTCTCTTGAAACGCATCTGTAC	46
Pro197GLN	CAATCATACGACG TT GGACTTGTCCTGTGATTGCTACAAGAGGAA
CCT-CAA	CACTATCTAACAACGCATCGGCTAATCCGCT
	ACAAGTCC AA CGTCGTA	47
	TACGACG TT GGACTTGT	48

Imidazolinone	GACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATGTGT	49
Resistance	TGCCGATGATCCCGA AC GGTGGCACTTTCAACGATGTCATAACGG
ALS	AAGGAGATGGCCGGATTAAATACTGAGAGAT
Arabidopsis thaliana	ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT	50
Ser653Asn	TGAAAGTGCCACC GT TCGGGATCATCGGCAACACATGTTCTTGGT
AGT-AAC	GCGGACAAATCACATCCAACAGGTAAGGTC
	GATCCCGA AC GGTGGCA	51
	TGCCACC GT TCGGGATC	52

Imidazolinone	GACCTTACCTGTTGGATGTGATTTGTCCGCACCAAGAACATGTGT	53
Resistance	TGCCGATGATCCCGA AT GGTGGCACTTTCAACGATGTCATAACGG
ALS	AAGGAGATGGCCGGATTAAATACTGAGAGAT
Arabidopsis thaliana	ATCTCTCAGTATTTAATCCGGCCATCTCCTTCCGTTATGACATCGT	54
Ser653Asn	TGAAAGTGCCACC AT TCGGGATCATCGGCAACACATGTTCTTGGT
AGT-AAT	GCGGACAAATCACATCCAACAGGTAAGGTC
	GATCCCGA AT GGTGGCA	55
	TGCCACC AT TCGGGATC	56

Sulfonylurea	TCCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGC	57
Resistance	CATCACGGGCCAGGTC T CCCGCCGCATGATCGGCACCGACGCCT
ALS	TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT
Oryza saliva	AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG	58
Pro171Ser	CCGATCATGCGGCGGG A GACCTGGCCCGTGATGGCGACCATCG
CCC-TCC	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA
	GCCAGGTC T CCCGCCGC	59
	GCGGCGGG A GACCTGGC	60

Sulfonylurea	CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC	61
Resistance	ATCACGGGCCAGGTCC AA CGCCGCATGATCGGCACCGACGCCTT
ALS	CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC
Oryza saliva	GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT 62
Pro171Gln	GCCGATCATGCGGCG TT GGACCTGGCCCGTGATGGCGACCATCG
CCC-CAA	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG
	CCAGGTCC AA CGCCGCA	63
	TGCGGCG TT GGACCTGG	64

Sulfonylurea	CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC	65
Resistance	ATCACGGGCCAGGTCC AG CGCCGCATGATCGGCACCGACGCCTT
ALS	CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC
Oryza saliva	GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT	66
Pro171Gln	GCCGATCATGCGGCG CT GGACCTGGCCCGTGATGGCGACCATCG
CCC-CAG	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG
	CCAGGTCC AG CGCCGCA	67
	TGCGGCG CT GGACCTGG	68

Imidazolinone	GGCCATACTTGTTGGATATCATCGTCCCGCACCAGGAGCATGTGC	69
Resistance	TGCCTATGATCCCAA A TGGGGGCGCATTCAAGGACATGATCCTGG
ALS	ATGGTGATGGCAGGACTGTGTATTAATCTAT
Oryza saliva	ATAGATTAATACACAGTCCTGCGATCACCATCCAGGATCATGTCCT	70
Ilee627Asn	TGAATGCGCCCCCA T TTGGGATCATAGGCAGCACATGCTCCTGGT
ATT-AAT	GCGGGACGATGATATCCAACAAGTATGGCC
	GATCCCAA A TGGGGGCG	71
	CGCCCCCA T TTGGGATC	72

Sulfonylurea	TCCGCGCTCGCCGACGCGCTGCTCGATTCCGTCCCCATGGTCGC	73
Resistance	CATCACGGGACAGGTG T CGCGACGCATGATTGGCACCGACGCCT
ALS	TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT
Zea mays	AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGGT	74
Pro165Ser	GCCAATCATGCGTCGCG A CACCTGTCCCGTGATGGCGACCATGG
CCG-TCG	GGACGGAATCGAGCAGCGCGTCGGCGAGCGCGGA
	GACAGGTG T CGCGACGC	75
	GCGTCGCG A CACCTGTC	76

Sulfonylurea	CCGCGCTCGCCGACGCGCTGCTCGATTCCGTCCCCATGGTCGCC	77
Resistance	ATCACGGGACAGGTGC A GCGACGCATGATTGGCACCGACGCCTT
ALS	CCAGGAGACGCCCATCGTCGAGGTCACCCGCTC
Zea mays	GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCGG	78
Pro165Gln	TGCCAATCATGCGTCGC T GCACCTGTCCCGTGATGGCGACCATG
CCG-CAG	GGGACGGAATCGAGCAGCGCGTCGGCGAGCGCGG
	ACAGGTGC A GCGACGCA	79
	TGCGTCGC T GCACCTGT	80

Imidazolinone	GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT	81
Resistance	TGCCTATGATCCCTA AT GGTGGGGCTTTCAAGGATATGATCCTGG
ALS	ATGGTGATGGCAGGACTGTGTACTGATCTAA
Zea mays	TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT	82
Ser621Asn	TGAAAGCCCCACC AT TAGGGATCATAGGCAACACATGCTCCTGGT
AGT-AAT	GTGGGACGATTATATCCAAGAGGTACGGCC
	GATCCCTA AT GGTGGGG	83
	CCCCACC AT TAGGGATC	84

Imidazolinone	GGCCGTACCTCTTGGATATAATCGTCCCACACCAGGAGCATGTGT	85
Resistance	TGCCTATGATCCCTA AC GGTGGGGCTTTCAAGGATATGATCCTGG
ALS	ATGGTGATGGCAGGACTGTGTACTGATCTAA
Zea mays	TTAGATCAGTACACAGTCCTGCCATCACCATCCAGGATCATATCCT	86
Ser621Asn	TGAAAGCCCCACC GT TAGGGATCATAGGCAACACATGCTCCTGGT
AGT-AAC	GTGGGACGATTATATCCAAGAGGTACGGCC
	GATCCCTA AC GGTGGGG	87
	CCCCACC GT TAGGGATC	88

Sulfonylurea	TCCGCGCTCGCCGACGCCGTCCTCGACTCCATCCCCATGGTGGC	89
Resistance	CATCACGGGGCAGGTC T CGCGCCGCATGATCGGCACGGACGCCT
ALS	TCCAGGAGACGCCCATCGTCGAGGTCACCCGCT
Lolium multiflorum	AGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCGTG	90
Pro167Ser	CCGATCATGCGGCGCG A GACCTGCCCCGTGATGGCCACCATGG
CCG-TCC	GGATGGAGTVGAGGAGGGCCTCGGCGACCCCCCA
	GGCAGGTC T CGCGCCGC	91
	GCGGCGCG A GACCTGCC	92

Sulfonylurea	CCGCGCTCGCCGACGCCCTCCTCGACTCCATCCCCATGGTGGCC	93
Resistance	ATCACGGGGCAGGTCC A GCGCCGCATGATCGGCACGGACGCCTT
ALS	CCAGGAGACGCCCATCGTCGAGGTCACCCGCTC
Lolium multiflorum	GAGCGGGTGACCTCGACGATGGGCGTCTCCTGGAAGGCGTCCGT	94
Pro167Gln	GCCGATCATGCGGCGC T GGACCTGCCCCGTGATGGCCACCATGG
CCG-CAG	GGATGGAGTCGAGGAGGGCGTCGGCGAGCGCGG
	GCAGGTCC A GCGCCGCA	95
	TGCGGCGC T GGACCTGC	96

Imidazolinone	CTGGGCCATACTTGTTGGATATCATCGTCCCTCACCAGGAGCATG	97
Resistance	TGCTGCCTATGATCCCTA A CGGTGGTGCTTTCAAGGACATTATCA
ALS	TGGAAGGTGATGGCAGGATTTCGTATTAAAC
Lolium multiflorum	GTTTAATACGAAATCCTGCCATCACCTTCCATGATAATGTCGTTGA	98
Ser623Asn	AAGCACCACCG T TAGGGATCATAGGCAGCACATGCTCCTGGTGA
AGC-AAC	GGGACGATGATATCCAACAAGTATGGCCCAG
	GATCCCTA A CGGTGGTG	99
	CACCACCG T TAGGGATC	100

Sulfonylurea	TCCGCGCTCGCCGACGGTCTCCTCGACTCCATCGCCATGGTCGC	101
Resistance	CATCACGGGCCAGGTC T CACGCCGCATGATCGGCACGGACGCGT
ALS	TCCAGGAGACGCCCATAGTGGAGGTCACGCGCT
Hordeum vulgare	AGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGTG	102
Pro68Ser	CGGATCATGCGGCGTG A GACCTGGCCCGTGATGGCGACCATGG
CCA-TCA	GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGGA
	GCCAGGTC T CACGCCGC	103
	GCGGCGTG A GACCTGGC	104

Sulfonyurea	CCGCGCTCGCCGACGCTCTCCTCGACTCCATCCCCATGGTCGCC	105
Resistance	ATCACGGGCCAGGTCC A ACGCCGCATGATCGGCACGGACGCGTT
ALS	CCAGGAGACGCCCATAGTGGAGGTCACGCGCTC
Hordeum vulgare	GAGCGCGTGACCTCCACTATGGGCGTCTCCTGGAACGCGTCCGT	106
Pro68Gln	GCCGATCATGCGGCGT T GGACCTGGCCCGTGATGGCGACCATGG
CCA-CAA	GGATGGAGTCGAGGAGAGCGTCGGCGAGCGCGG
	CCAGGTCC A ACGCCGCA	107
	TGCGGCGT T GGACCTGG	108

Imidazolinone	CCCAGGGCCGTACCTGCTGGATATCATTGTCCCGCATCAGGAGC	109
Resistance	ACGTGCTGCCTATGATCCCAA A CGGTGGTGCTTTCAAGGACATGA
ALS	TCATGGAGGGTGATGGCAGGACCTCGTACTGA
Hordeum vulgare	TCAGTACGAGGTCCTGCCATTCACCCTCCATGATCATGTCCTTGAA	110
Ser524Asn	AGCACCACCG T TTGGGATCATAGGCAGCACGTGCTCCTGATGCG
AGC-AAC	GGACAATGATATCCAGCAGGTACGGCCCTGGG
	GATCCCAA A CGGTGGTG	111
	CACCACCG T TTGGGATC	112

Sulfonylurea	AGTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCG	113
Resistance	ATCACTGGTCAAGTC T CTCGTCGGATGATCGGTACCGATGCTTTC
ALS	CAGGAAACTCCAATTGTTGAGGTAACAAGGT
Gossypium hirsutum	ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC	114
Pro186Ser	CGATCATCCGACGAG A GACTTGACCAGTGATCGCCACGAGAGGG
CCT-TCT	ATACTATCGAGCATTGCATCAGCGAGACCACT
	GTCAAGTC T CTCGTCGG	115
	CCGACGAG A GACTTGAC	116

Sulfonylurea	GTGGTCTCGCTGATGCAATGGTCGATAGTATCCCTCTCGTGGCGA	117
Resistance	TCACTGGTCAAGTCC AA CGTCGGATGATCGGTACCGATGCTTTCC
ALS	AGGAAACTCCAATTGTTGAGGTAACAAGGTC
Gossypium hirsutum	GACCTTGTTACCTCAACAATTGGAGTTICCTGGAAAGCATCGGTA	118
Pro186Gln	CCGATCATCCGACG TT GGACTTGACCAGTGATCGCCACGAGAGG
CCT-CAA	GATACTATCGAGCATTGCATCAGCGAGACCAC
	TCAAGTCC AA CGTCGGA	119
	TCCGACG TT GGACTTGA	120

Sulfonylurea	GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCGA	121
Resistance	TCACIGGTCAAGTCC AG CGTCGGATGATCGGTACCGATGCTTTCC
ALS	AGGAAACTCCAATTGTTGAGGTAACAAGGTC
Gossypium hirsutum	GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA	122
Pro186Gln	CCGATCATCCGACG CT GGACTTGACCAGTGATCGCCACGAGAGG
CCT-CAG	GATACTATCGAGCATTGCATCAGCGAGACCAC
	TCAAGTCC AG CGTCGGA	123
	TCCGACG CT GGACTTGA	124

Imidazolinone	GACCTTACTTGTTGGATGTGATTGTCCCACATCAAGAACATGTCCT	125
Resistance	GCCTATGATCCCCA A TGGAGGCGCTTTCAAAGATGTGATCACAGA
ALS	GGGTGATGGAAGAACACAATATTGACCTCA
Gossypium hirsutum	TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT	126
Ser642Asn	TGAAAGCGCCTCCA T TGGGGATCATAGGCAGGACATGTTCTTGAT
AGT-AAT	GTGGGACAATCACATCCAACAAGTAAGGTC
	GATCCCCA A TGGAGGCG	127
	CGCCTCCA T TGGGGATC	128

Sulfonylurea	TCTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCA	129
Resistance	TTACTGGGCAAGTT T CCCGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGAT
Amaranthus	ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC	130
retroflexus	AATCATACGCCGGG A AACTTGCCCAGTAATGGCGACAAGAGGGA
Pro192Ser	CTGAGTCAAGAAGTGCATCAGCAAGACCAGA
CCC-TCC	GGCAAGTT T CCCGGCGT	131
	ACGCCGGG A AAGTTGCC	132

Sulfonylurea	CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT	133
Resistance	TACTGGGCAAGTTC AA CGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGATC
Amaranthus	GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC	134
retroflexus	CAATCATACGCCG TT GAACTTGCCCAGTAATGGCGACAAGAGGGA
Pro192Gln	CTGAGTCAAGAAGTGCATCAGCAAGACCAG
CCC-CAA	GCAAGTTC AA CGGCGTA	135
	TACGCCG TT GAACTTGC	136

Sulfonylurea	CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT	137
Resistance	TACTGGGCAAGtTC AG CGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGATC
Amaranthus	GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC	138
retroflexus	CAATCATACGCCG CT GAACTTGCCCAGTAATGGCGACAAGAGGG
Pro192Gln	ACTGAGTCAAGAAGTGCATCAGCAAGACCAG
CCC-CAG	GCAAGTTC AG CGGCGTA	139
	TACGCCG CT GAACTTGC	140

Imidazolinone	GACCGTATCTTGCTGGATGTTAATCGTACCACATCAGGAGCATGTGC	141
Resistance	TGCCTAIGATCCCTA A CGGTGCCGCCTTCAAGGACACCATAACAG
ALS	AGGGTGATGGAAGAAGGGGTTATTAGTTGGT
Amaranthus	ACCAACTAATAAGCCCTTCTTCCATTCACCCTCTGTTATGGTGTCCT	142
retroflexus	TGAAGGCGGCACCG T TAGGGATCATAGGCAGCACATGCTCCTGA
Ser652Asn	TGTGGTACGATTACATCCAGCAGATACGGTC
AGC-AAC	GATCCCTA A CGGTGCCG	143
	CGGCACCG T TAGGGATC	144

Sulfonylurea	AGCGGCCTCGCTGACGCGCTACTGGATAGCGTCCCCATTGTTGC	145
Resistance	TATAACAGGTCAAGTG T CACGTAGGATGATAGGTACTGATGCTTTT
ALS 1	CAGGAAACTCCTATTGTITGAGGTAACTAGAT
Nicotiana tabacum	ATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTACC	146
Pro194Ser	TATCATCCTACGTG A CACTTGACCTGTTATAGCAACAATGGGGAC
CCA-TCA	GCTATCCAGTAGCGCGTCAGCGAGGCCGCT
	GTCAAGTG T CACGTAGG	147
	CCTACGTG A CACTTGAC	148

Sulfonylurea	GCGGCCTCGCTGACGCGCTACTGGATAGCGTCCCCATTGTTGCT	149
Resistance	ATAACAGGTCAAGTGC AA CGTAGGATGATAGGTACTGATGCTTTT
ALS 1	CAGGAAACTCCTATTGTTGAGGTAACTAGATC
Nicotiana tabacum	GATCTAGTTACCTCAACAATAGGAGTTTCCTGAAAAGCATCAGTAC	150
Pro194Gln	CTATCATCCTACGT T GCACTTGACCTGTTATAGCAACAATGGGGA
CCA-CAA	CGCTATCCAGTAGCGCGTCAGCGAGGCCGC
	TCAAGTGC A ACGTAGGA	151
	TCCTACGT T GCACTTGA	152

Imidazolinone	GGCCATACTTGTTGGATGTGATTGTACCTCATCAGGAACATGTTTT	153
Resistance	ACCTATGATTCCCA A TGGCGGAGCTTTCAAAGATGTGATCACAGA
ALS 1	GGGTGACGGGAGAAGTTCCTATTGAGTTTG
Nicotiana tabacum	CAAACTGAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT	154
Ser650Asn	TGAAAGCTCCGCCA T TGGGAATCATAGGTAAAACATGTTCCTGAT
AGT-AAT	GAGGTACAATCACATCCAACAAGTATGGCC
	GATTCCCA A TGGCGGAG	155
	CTCCGCCA T TGGGAATC	156

Sulfonylurea	AGTGGCCTCGCGGACGCCCTACTGGATAGCGTCCCCATTGTTGC	157
Resistance	TATAACCGGTCAAGTG T CACGTAGGATGATCGGTACTGATGCTTT
ALS 2	TCAGGAAACTCCGATTGTTGAGGTAACTAGAT
Nicotiana tabacum	ATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTACC	158
Pro191Ser	GATCATCCTACGTG A CACTTGACCGGTTATAGCAACAATGGGGAC
CCA-TCA	GCTATCCAGTAGGGCGTCCGCGAGGCCACT
	GICAAGTG T CACGTAGG	159
	CCTACGTG A CACTTGAC	160

Sulfonylurea	GTGGCCTCGCGGACGCCCTACTGGATAGCGTCCCCATTGTTGCT	161
Resistance	ATAACCGGTCAAGTGC A ACGTAGGATGATCGGTACTGATGCTTTT
ALS 2	CAGGAAACTCCGATTGTTGAGGTAACTAGATC
Nicotiana tabacum	GATCTAGTTACCTCAACAATCGGAGTTTCCTGAAAAGCATCAGTAC	162
Pro191Gln	CGATCATCCTACGT T GCACTTGACCGGTTATAGCAACAATGGGGA
CCA-CAA	CGCTATCCAGTAGGGCGTCCGCGAGGCCAC
	TCAAGTGC A ACGTAGGA	163
	TCCTACGT T GCACTTGA	164

Imidazolinone	GGCCATACTTGTTGGATGTGATTGTACCTCATCAGGAACATGTTCT	165
Resistance	ACCTATGATTCCCA A TGGCGGGGCTTTCAAAGATGTGATCACAGA
ALS 2	GGGTGACGGGAGAAGTTCCTATTGACTTTG
Nicotiana tabacum	CAAAGTCAATAGGAACTTCTCCCGTCACCCTCTGTGATCACATCTT	166
Ser647Asn	TGAAAGCCCCGCCA T TGGGAATCATAGGTAGAACATGTTCCTGAT
AGT-AAT	GAGGTACAATCACATCCAACAAGTATGGCC
	GATTCCCA A TGGCGGGG	167
	CCCCGCCA T TGGGAATC	168

Sulfonylurea	AGTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTA	169
Resistance	TTACTGGTCAAGTT T CCAGGAGAATGATTGGAACAGATGCGTTTC
ALS	AAGAAACCCCTATTGTTGAGGTAACACGTT
Xanthium spp.	AACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTCC	170
Pro175Ser	AATCATTCTCCTGG A AACTTGACCAGTAATAGCAACCATTGGAACA
CCC-TCC	CTGTCTAATAAAGCATCAGCAAGACCACT
	GTCAAGTT T CCAGGAGA	171
	TCTCCTGG A AACTTGAC	172

Sulfonylurea	GTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT	173
Resistance	TACTGGTCAAGTTC AA AGGAGAATGATTGGAACAGATGCGTTTCA
ALS	AGAAACCCCTATTGTTGAGGTAACACGTTC
Xanthium spp.	GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC	174
Pro175Gln	CAATCATTCTCCT TT GAACTTGACCAGTAATAGCAACCATTGGAAC
CCC-CAA	ACTGTCTAATAAAGCATCAGCAAGACCAC
	TCAAGTTC AA AGGAGAA	175
	TTCTCCT TTGAACTTGA	176

Sulfonylurea	GTGGTCTTGCTGATGCTTTATTAGACAGTGTTCCAATGGTTGCTAT	177
Resistance	TACTGGTCAAGTTC AGAGGAGAATGATTGGAACAGATGCGTTTCA
ALS	AGAAACCCCTATTGTTGAGGTAACACGTTC
Xanthium spp.	GAACGTGTTACCTCAACAATAGGGGTTTCTTGAAACGCATCTGTTC 178
Pro175Gln	CAATCATTCTCCT CT GAACTTGACCAGTAATAGCAACCATTGGAAC
CCC-CAG	ACTGTCTAATAAAGCATCAGCAAGACCAC
	TCAAGTTC AG AGGAGAA	179
	TTCTCCT CT GAACTTGA	180

Imidazolinone	GGGCCTTACTTGTTGGATGTGATCGTGCCCCATCAAGAACATGTG	181
Resistance	TTGCCCATGATCCCG AA TGGTGGAGGTTTCATGGATGTGATCACC
ALS	GAAGGCGACGGCAGAATGAAATATTGAGCTT
Xanthium spp.	AAGCTCAATATTTCATTCTGCCGTCGCCTTCGGTGATCACATCCAT	182
Ala631Asn	GAAACCTCCACCA TT CGGGATCATGGGCAACACATGTTCTTGATG
GCT-AAT	GGGCACGATCACATCCAACAAGTAAGGCCC
	TGATCCCG AA TGGTGGA	183
	TCCACCA TT CGGGATCA	184

Sulfonylurea	TCCGGGTTTGCTGATGCTTTGCTCGATTCCGTTCCACTGGTGGCG	185
Resistance	ATCACGGGGCAGGTG T CGCGGCGAATGATTGGGACGGATGCTTT
ALS	TCAGGAGACTCCTATTGTTGAGGTAACACGGT
Bassia scoparia	ACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTCC	186
Pro189Ser	CAATCATTCGCCGCG A CACCTGCCCCGTGATCGCCACCAGTGGA
CCG-TCG	ACGGAATCGAGCAAAGCATCAGCAAACCCGGA
	GGCAGGTG T CGCGGCGA	187
	TCGCCGCG A CACCTGCC	188

Sulfonylurea	CCGGGTTTGGTGATGCTTTGCTCGATTCCGTTCCACTGGTGGCGA	189
Resistance	TCACGGGGCAGGTGC A GCGGCGAATGATTGGGACGGATGCTTTT
ALS	CAGGAGACTCCTATTGTTGAGGTAACACGGTC
Bassia scoparia	GACCGTGTTACCTCAACAATAGGAGTCTCCTGAAAAGCATCCGTC	190
Pro189Gln	CCAATCATTCGCCGC T GCACCTGCCCCGTGATCGCCACCAGTGG
CCG-CAG	AACGGAATCGAGCAAAGCATCAGCAAACCCGG
	GCAGGTGC A GCGGCGAA	191
	TTCGCCGC T GCAGCTGC	192

Imidazolinone	GACCTTACCTGCTTGATGTGATTGTACCTCATCAGGAGCATGTGC	193
Resistance	TGCCTATGATTCCTA A TGGTGCAGCCTTCAAGGATATCATTAACGA
ALS	AGGTGATGGAAGAACAAGTTATTGATGTTC
Bassia scoparia	GAACATCAATAACTTGTTCTTCCATCACCTTCGTTAATGATATCCTT	194
Ser649Asn	GAAGGCTGCACCA T TAGGAATCATAGGCAGCACATGCTCCTGATG
AGT-AAT	AGGTACAATCACATCAAGCAGGTAAGGTC
	GATTCGTA A TGGTGCAG	195
	CTGCACCA T TAGGAATC	196

Sulfonylurea	AGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCC	197
Resistance	ATTACAGGACAGGTC T CTCGCCGGATGATCGGTACTGACGCCTTC
ALS 1	CAAGAGACACCAATCGTTGAGGTAACGAGGT
Brassica napus	ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC	198
Pro182Ser	CGATCATCCGGCGAG A GACCTGTCCTGTAATGGCGACAAGAGGA
CCT-TCT	ACACTGTCAAGCATCGCGTCTGCTAACCCGCT
	GACAGGTC T CTCGCCGG	199
	CCGGCGAG A GACCTGTC	200

Sulfonylurea	GCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCCA	201
Resistance	TTACAGGACAGGTCC AA CGCCGGATGATCGGTACTGACGCCTTC
ALS 1	CAAGAGACACCAATCGTTGAGGTAACGAGGTC
Brassica napus	GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA	202
Pro182Gln	CCGATCATCCGGCG TT GGACCTGTCCTGTAATGGCGACAAGAGG
CCT-CAA	AACACTGTCAAGCATCGCGTCTGCTAACCCGC
	ACAGGTCC AA CGCCGGA	203
	TCCGGCG TT GGACCTGT	204

Sulfonylurea	GCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCCA	205
Resistance	TTACAGGACAGGTCC AG CGCCGGATGATCGGTACTGACGCCTTC
ALS 1	CAAGAGACACCAATCGTTGAGGTAACGAGGTC
Brassica napus	GACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTA	206
Pro182Gln	CCGATCATCCGGCG CT GGACCTGTCCTGTAATGGCGACAAGAGG
CCT-CAG	AACACTGTCAAGCATCGCGTCTGCTAACCCGC
	ACAGGTCC AG CGCCGGA	207
	TCCGGCG CT GGACCTGT	208

Imidazolinone	GACCATACCTGTTGGATGTGATATGTCCGCACCAAGAACATGTGT	209
Resistance	TACCGATGATCCCAA A TGGTGGCACTTTCAAAGATGTAATAACAG
ALS 1	AAGGGGATGGTCGCACTAAGTACTGAGAGAT
Brassica napus	ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT	210
Ser638Asn	GAAAGTGCCACCA T TTGGGATCATCGGTAACACATGTTCTTGGTG
AGT-AAT	CGGACATATCACATCCAACAGGTATGGTC
	GATCCCAA A TGGTGGCA	211
	TGCCACCA T TTGGGATC	212

Sulfonylurea	CAGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGC	213
Resistance	CATTACAGGACAGGT T CCTCGCCGGATGATCGGTACTGACGCCTT
ALS 2	CCAAGAGACACCAATCGTTGAGGTAACGAGG
Brassica napus	CCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTACC	214
Pro126Ser	GATCATCCGGCGAGG A ACCTGTCCTGTAATGGCGACAAGAGGAA
CCC-TCC	CACTGTCAAGCATCGCGTCTGCTAACCCGCTG
	GGACAGGT T CCTCGCCG	215
	CGGCGAGG A ACCTGTCC	216

Sulfonylurea	AGCGGGTTAGCAGACGCGATGCTTGACAGTGTTCCTCTTGTCGCC	217
Resistance	ATTACAGGACAGGTC A CTCGCCGGATGATCGGTACTGACGCCTTC
ALS 2	CAAGAGACACCAATCGTTGAGGTAACGAGGT
Brassica napus	ACCTCGTTACCTCAACGATTGGTGTCTCTTGGAAGGCGTCAGTAC	218
Pro126Gln	CGATCATCCGGCGAG T GACCTGTCCTGTAATGGCGACAAGAGGA
CCC-CAG	ACACTGTCAAGCATCGCGTCTGCTAACCCGCT
	GACAGGTC A CTCGCCGG	219
	CCGGCGAG T GACCTGTC	220

Imidazolinone	GACCATACCTGTTGGATGTGATATGTCCGCACCAAGAACATGTGT	221
Resistance	TACCGATGATCCCAA A TGGTGGCACTTTCAAAGATGTAATAACAG
ALS 2	AAGGGGATGGTCGCACTAAGTACTGAGAGAT
Brassica napus	ATCTCTCAGTACTTAGTGCGACCATCCCCTTCTGTTATTACATCTTT	222
Ser582Asn	GAAAGTGCCACCA T TTGGGATCATCGGTAACACATGTTCTTGGTG
AGT-AAT	CGGACATATCACATCCAACAGGTATGGTC
	GATCCCAA A TGGTGGCA	223
	TGCCACCA T TTGGGATC	224

Sulfonylurea	AGCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGC	225
Resistance	CATCACAGGACAGGTC T CTCGCCGGATGATCGGTACTGACGCGT
ALS 3	TCCAAGAGACGCCAATCGTTGAGGTAACGAGGT
Brassica napus	ACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTAC	226
Pro179Ser	CGATCATCCGGCGAG A GACCTGTCCTGTGATGGCGACGAGAGGA
CCT-TCT	ACACTGTCAAGCATCGCGTCGGCTAACCCGCT
	GACAGGTC T CTCGCCGG	227
	CCGGCGAG A GACCTGTC	228

Sulfonylurea	GCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGCC	229
Resistance	ATCACAGGACAGGTCC AA CGCCGGATGATCGGTACTGACGCGTT
ALS 3	CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC
Brassica napus	GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA	230
Pro179Gln	CCGATCATCCGGCG TT GGACCTGTCCTGTGATGGCGACGAGAGG
CCT-CAA	AACACTGTCAAGCATCGCGTCGGCTAACCCGC
	ACAGGTCC AAee CGCCGGA	231
	TCCGGCG TT GGACCTGT	232

Sulfonylurea	GCGGGTTAGCCGACGCGATGCTTGACAGTGTTCCTCTCGTCGCC	233
Resistance	ATCACAGGACAGGTCC AG CGCCGGATGATCGGTACTGACGCGTT
ALS 3	CCAAGAGACGCCAATCGTTGAGGTAACGAGGTC
Brassica napus	GACCTCGTTACCTCAACGATTGGCGTCTCTTGGAACGCGTCAGTA	234
Pro179Gln	CCGATCATCCGGCG CT GGACCTGTCCTGTGATGGCGACGAGAGG
CCT-CAG	AACACTGTCAAGCATCGCGTCGGCTAACCCGC
	ACAGGTCC AG CGCCGGA	235
	TCCGGCG CT GGACCTGT	236

Imidazolinone	GACCGTACCTGTTGGATGTCATCTGTCCGCACCAAGAACATGTGT	237
Resistance	TACOGATGATCCCAA A TGGTGGCACTTTCAAAGATGTAATAACCG
ALS 3	AAGGGGATGGTCGCACTAAGTACTGAGAGAT
Brassica napus	ATCTCTCAGTACTTAGTGCGACCATCCCCTTCGGTTATTACATCTT	238
Ser635Asn	TGAAAGTGCCACCA T TTGGGATCATCGGTAACACATGTTCTTGGT
AGT-AAT	GCGGACAGATGACATCCAACAGGTACGGTC
	GATCCCAA A TGGTGGCA	239
	TGCCACCA T TTGGGATC	240

Sultonylurea	TCCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGC	241
Resistance	CATCACGGGCCAGGTC T CCCGCCGCATGATCGGCACCGACGCCT
ALS	TCCAGGAGACGCCCATAGTCGAGGTCACCCGCT
Oryza sativa	AGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGTG	242
Prol7l Ser	CCGATCATGCGGCGGG A GACCTGGCCCGTGATGGCGACCATCG
CCC-TCC	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGGA
	GCCAGGTC T CCCGCCGC	243
	GCGGCGGG A GACCTGGC	244

Sulfonylurea	CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC	245
Resistance	ATCACGGGCCAGGTCC AA CGCCGCATGATCGGCACCGACGCCTT
ALS	CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC
Oryza sativa	GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT	246
Pro171Gln	GCCGATCATGCGGCG Tee TGGACCTGGCCCGTGATGGCGACCATCG
CCC-CAA	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG
	CCAGGTCC AA CGCCGCA	247
	TGCGGCG TT GGACCTGG	248

Sulfonylurea	CCGCGCTCGCCGACGCGCTGCTCGACTCCGTCCCGATGGTCGCC	249
Resistance	ATCACGGGCCAGGTCC AG CGCCGCATGATCGGCACCGACGCCTT
ALS	CCAGGAGACGCCCATAGTCGAGGTCACCCGCTC
Oryza sativa	GAGCGGGTGACCTCGACTATGGGCGTCTCCTGGAAGGCGTCGGT 250
Pro171Gln	GCCGATCATGCGGCG CT GGACCTGGCCCGTGATGGGGACCATCG
CCC-CAG	GGACGGAGTCGAGCAGCGCGTCGGCGAGCGCGG
	CCAGGTCC AG CGCCGCA	251
	TGCGGCG CT GGACCTGG	252

Imidazolinone	GGCCATACTTGTTGGATATCATCGTCCCGCACCAGGAGCATGTGC	253
Resistance	TGCCTATGATCCCAA A TGGGGGCGCATTCAAGGACATGATCCTGG
ALS	ATGGTGATGGCAGGACTGTGTATTAATCTAT
Oryza sativa	ATAGATTAATACACAGTCCTGCCATCACCATCCAGGATCATGTCCT	254
Ser627Asn	TGAATGCGCCCCCA T TTGGGATCATAGGCAGCACATGCICCTGGI
AGT-AAT	GCGGGACGATGATATCCAACAAGTATGGCC
	GATCCCAA A TGGGGGCG	255
	CGCCCCGA T TTGGGATC	256

Sulfonylurea	TCTGCGCTCGCAGACGCGTTGCTCGACTCCGTCCCCATGGTCGC	257
Resistance	CATCACGGGACAGGTG T CGCGACGCATGATTGGCACCGACGCCT
ALS	TTCAGGAGACGCCCATCGTCGAGGTCACCCGCT
Zea mays	AGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGTG	258
Pro165Ser	CCAATCATGCGTCGCG A CACCTGTCCCGTGATGGCGACCATGGG
CCG-TCG	GACGGAGTCGAGCAACGCGTCTGCGAGCGCAGA
	GACAGGTG T CGCGACGC	259
	GCGTCGCG A CACCTGTC	260

Sulfonylurea	CTGCGCTCGCAGACGCGTTGCTCGACTCCGTCCCCATGGTCGCC	261
Resistance	ATCACGGGACAGGTGC A GCGACGCATGATTGGCACCGACGCCTT
ALS	TCAGGAGACGCCCATCGTCGAGGTCACCCGCTC
Zea mays	GAGCGGGTGACCTCGACGATGGGCGTCTCCTGAAAGGCGTCGGT 262
Pro165Gln	GCCAATCATGCGTCGC T GCACCTGTCCCGTGATGGCGACCATGG
CCG-CAG	GGACGGAGTCGAGCAACGCGTCTGCGAGCGCAG
	ACAGGTGC A GCGACGCA	263
	TGCGTCGC T GCACCTGT	264

Imidazolinone	GGCCGTACCTCTTGGATATAATCGTCCCGCACCAGGAGCATGTGT	265
Resistance	TGCCTATGATCCCTA A TGGTGGGGCTTTCAAGGATATGATCCTGG
ALS	ATGGTGATGGCAGGACTGTGTATTGATCCGT
Zea mays	ACGGATCAATACACAGTCCTGCCATCACCATCCAGGATCATATCC	266
Ser621Asn	TTGAAAGCCCCACCA T TAGGGATCATAGGCAACACATGCTCCTGG
AGT-AAT	TGCGGGACGATTATATCCAAGAGGTACGGCC
	GATCCCTA A TGGTGGGG	267
	CCCCACCA T TAGGGATC	268

Sulfonylurea	AGTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCG	269
Resistance	ATCACTGGICAAGTC T CTCGTCGGATGATCGGTACCGATGCTTTC
ALS	CAGGAAACTCCAATTGTTGAGGTAACAAGGT
Gossypium hirsutum	ACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTAC	270
Pro186Ser	CGATCATCCGACGAG A GACTTGACCAGTGATCGCCACGAGAGGG
CCT-TCT	ATACTATGGAGCATTGCATCAGCGAGACCACT
	GTCAAGTCTC T CGTCGG	271
	CCGACGAGAG A CTTGAC	272

Sulfonylurea	GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGGCGA	273
Resistance	TCACTGGTCAAGTCC AA CGTCGGATGATCGGTACCGATGCTTTCC
ALS	AGGAAACTCCAATTGTTGAGGTAACAAGGTC
Gossypium hirsutum	GACCTTGTTACCTTAACAATTGGAGTTTCCTGGAAAGCATCGGTA	274
Pro186Gln	CCGATCATCCGACG TT GGACTTGACCAGTGATCGCCACGAGAGG
CCT-CAA	GATACTATCGAGCATTGCATCAGCGAGACCAC
	TCAAGTCC AA CGTCGGA	275
	TTCCGACG TT GGACTTGA	276

Sulfonylurea	GTGGTCTCGCTGATGCAATGCTCGATAGTATCCCTCTCGTGCCGA	277
Resistance	TCACTGGTCAAGTCC AG CGTCGGATGATCGGTACCGATGCTTTCC
ALS	AGGAAACTCCAATTGTTGAGGTAACAAGGTC
Gossypium hirsutum	GACCTTGTTACCTCAACAATTGGAGTTTCCTGGAAAGCATCGGTA	278
Pro186Gln	CCGATCATCCGACG CT GGACTTGACCAGTGATCGCCACGAGAGG
CCT-CAG	GATACTATCGAGCATTGCATCAGCGAGACCAC
	TCAAGTCC AG CGTCGGA	279
	TCCGACG CT GGACTTGA	280

Imidazolinone	GACCTTACTTGTTGGATGTGATTGTCCCACATCAAGAACATGTCCT	281
Resistance	GCCTATGATCCCCA A TGGAGGGGCTTTCAAAGATGTGATCACAGA
ALS	GGGTGATGGAAGAACACAATATTGACCTCA
Gossypium hirsutum	TGAGGTCAATATTGTGTTCTTCCATCACCCTCTGTGATCACATCTT	282
Ser642Asn	TGAAAGCCCCTCCA T TGGGGATCATAGGCAGGACATGTTCTTGAT
AGT-AAT	GTGGGACAATCACATCCAACAAGTAAGGTC
	GATCCCCA A TGGAGGGG	283
	CCCCTCCA Tee TGGGGATC	284

Sulfonylurea	TCTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCA	285
Resistance	TTACTGGGCAAGTT T CCCGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGAT
Amaranthus powellii	ATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTACC	286
Pro192Ser	AATCATACGCCGGG A AACTTGCCCAGTAATGGCGACAAGAGGGA
CCC-TCC	CTGAGTCAAGAAGTGCATCAGCAAGACCAGA
	GGCAAGTT T CCCGGCGT	287
	ACGCCGGG A AACTTGCC	288

Sulfonymurea	CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT	289
Resistance	TACTGGGC AA GTTCAACGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGATC
Amaranthus powellii	GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC	290
Pro192Gln	CAATCATACGCCG TT GAACTTGCCCAGTAATGGCGACAAGAGGGA
CCC-CAA	CTGAGTCAAGAAGTGCATCAGCAAGACCAG
	GCAAGTTC AA CGGCGTA	291
	TACGCCG TT GAACTTGC	292

Sulfonylurea	CTGGTCTTGCTGATGCACTTCTTGACTCAGTCCCTCTTGTCGCCAT	293
Resistance	TACTGGGCAAGTTC AG CGGCGTATGATTGGTACTGATGCTTTTCA
ALS	AGAGACTCCAATTGTTGAGGTAACTCGATC
Amaranthus powellii	GATCGAGTTACCTCAACAATTGGAGTCTCTTGAAAAGCATCAGTAC	294
Pro192Gln	CAATCATACGCCG CT GAACTTGCCCAGTAATGGCGACAAGAGGG
CCC-CAG	ACTGAGTCAAGAAGTGCATCAGCAAGACCAG
	GCAAGTTC AG CGGCGTA	295
	TACGCCG CT GAACTTGC	296

Imidazolinone	GACCGTATCTGCTGGATGTAATCGTACCACATCAGGAGCATGTGC	297
Resistance	TGCCTATGATCCCTA A CGGTGCCGCCTTCAAGGACACCATAACAG
ALS	AGGGTGATGGAAGAAGGGCTTATTAGTTGGT
Amaranthus powellii	ACCAACTAATAAGCCCTTCTTCCATCACCCTCTGTTATGGIGTCCT	298
Ser652Asn	TGAAGGCGGCACCG T TAGGGATCATAGGCAGCACATGCTCCTGA
AGC-AAC	TGTGGTACGATTACATCCAGCAGATACGGTG
	GATCCCTA A CGGTGCCG	299
	CGGCACCG T TAGGGATC	300

TABLE 12


Genome-Altering Oligos Conferring Porphyric Herbicide Resistance

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Porphyric Herbicide	TCTTGCGCCCTCTTTCTGAATCTGCTGCAAATGCACTCTCAAAACT	301
Resistant	ATATTACCCACCA ATG GCAGCAGTATCTATCTCGTACCCGAAAGA
PPO	AGCAATCCGAACAGAATGTTTGATAGATGG
Arabidopsis thaliana	CCATCTATCAAACATTCTGTTCGGATTGCTTCTTTCGGGTACGAGA	302
Val365Met	TAGATACTGCTG C CA T TGGTGGGTAATATAGTTTTGAGAGTGCATT
GTT-ATG	TGCAGCAGATTCAGAAAGAGGGCGCAAGA
	CCCACCA A T G GCAGCAG	303
	CTGCTGC C A T TGGTGGG	304

Porphyric Herbicide	TATTACGTCCTCTTTCGGTTGCCGCAGCAGATGCACTTTCAAATTT	305
Resistant	CTACTAICCCCCA A T G GGAGCAGTCACAATTTCATATCCTCAAGAA
PPO	GCTATTCGTGATGAGCGTCTGGTTGATGG
Nicotiana tabacum	CCATCAACCAGACGCTCATCACGAATAGCTTCTTGAGGATATGAA	306
Val376Met	ATTGTGACTGCTCC C A TT GGGGGATAGTAGAAATTTGAAAGTGCA
GTT-ATG	TCTGCTGCGGCAACCGAAAGAGGACGTAATA
	TCCCCCA A T GGGAGCAG	307
	CTGCTCC C A T TGGGGGA	308

Porphyric Herbicide	TGTTGCGTCCGCTTTCGTTGGGTGCAGCAGATGCATTGTCAAAAT	309
Resistant	TTTATTATCCTCCG A T G GCAGCTGTATCAATTTCATATCCAAAAGA
PPO	CGGAATTCGTGCTGACCGGCTGATTGATGG
Cichorium intybus	CCATCAATCAGCCGGTCAGCACGAATTGCGTCTTTTGGATATGAA	310
Val383Met	ATTGATACAGCTGC C A T CGGAGGATAATAAAATTTTGACAATGCAT
GTT-ATG	CTGCTGCACCCAACGAAAGCGGACGCAACA
	TCCTCCG A T GGCAGCTG	311
	CAGCTGC C A T CGGAGGA	312

Porphyric Herbicide	TCCTTCGTCCACTTTCAGATGTCGCCGCAGAATCTCTTTCAAAATT	313
Resistant	TCATTATCCACCA A T G GCAGCTGTGTCACTTTCCTATCCTAAAGAA
PPO	GCAATTAGATCAGAGTGCTTGATTGACGG
Spinacia oleracea	CCGTCAATCAAGCACTCTGATCTAATTGCTTCTTTAGGATAGGAAA	314
Val390Met	GTGACACAGCTGC C A T TGGTGGATAATGAAATTTTGAAAGAGATT
GTT-ATG	CTGCGGCGACATCTGAAAGTGGACGAAGGA
	TCCACCA A T G GCAGCTG	315
	CAGCTGC C A T TGGTGGA	316

Porphyric Herbicide	TTTTGCGTCCACTTTCAAGCGATGCTGCAGATGCTCTATCAAGATT	317
Resistant	CTATTATCCACCG A T G GCTGCIGTAACTGTTTCGTATCCAAAGGAA
PPO	GCAATTAGAAAAGAATGCTTAATTGATGG
Zea mays	CGATCAATTAAGCATTCTTTTCTAATTGCTTCCTTTGGATACGAAAC	318
Val363Met	AGTTACAGCAGC C A T CGGTGGATAATAGAATCTTGATAGAGCATC
GTT-ATG	TGCAGCATCGCTTGAAAGTGGACGCAAAA
	TCCACCG A T G GCTGCTG	319
	CAGCAGC C A T CGGTGGA	320

Porphyric Herbicide	TCTTGCGGCCACTTTCAAGTGATGGAGCAGATGCTCTGTCAATATT	321
Resistant	CTATTATCCACCA A T G GCTGCTGTAACTGTTTCATATCCAAAAGAA
PPO	GCAATTAGAAAAGAATGCTTAATTGACGG
Oryza sativa	CCGTCAATTAAGCATTCTTTTCTAATTGCTTCTTTTGGATATGAAAC	322
Val364Met	AGTTACAGCAGC C A T TGGTGGATAATAGAATATTGACAGAGCATC
GTT-ATG	TGCTGCATCACTTGAAAGTGGCCGCAAGA
	TCCACCA A T G GCTGCTG	323
	CAGCAGCCA T TGGTGGA	324

Porphyric Herbicide	CTGGTCAAGGAGCAGGCGCCCGCCGCCGCCGAGGCCCTGGGCT	325
Resistant	CCTTCGACTACCCGCCG A TGGGCGCCGTGACGCTGTCGTACCCG
PPO	CTGAGCGCCGTGCGGGAGGAGCGCAAGGCCTCGG
Chlamydomonas	CCGAGGCCTTGCGCTCCTCCCGCACGGCGCTCAGCGGGTACGAC	326
reinhardtii	AGCGTCACGGCGCCCA T CGGCGGGTAGTCGAAGGAGCCCAGGG
Val389Met	CCTCGGCGGCGGCGGGCGCCTGCTCCTTGACCAG
GTG-ATG	ACCCGCCG A TGGGCGCC	327
	GGCGCCCA T CGGGGGGT	328

TABLE 13


Genome-Altering Oligos Conferring Triazine Resistance

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	329
D1 Protein	TTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCTT
Arabidopsis thaliana	AGCGGCTTGGCCGGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	330
AGT-ACT	CGAGAATIGTTGAAA G TAGCATATTGGAAAATCAATCGGCCAAAAT
	AACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCTA C TTTCAACA	331
	TGTTGAAA G TAGCATAT	332
Triazine Resistant	AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	333
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Nicotiana tabacum	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAG GCCAAGCAGCTAGGAAGAAGTGTAACGAA	334
AGT-ACT	CGAGAGtTGTIGAAA G TAGCATATTGGAAGATCAAtCGGCCAAAA
	TAACCATGAGCGGCTACGATGTTATAAGTTT
	ATATGCTA C TTTCAACA	335
	TGTTGAAA G TAGCATAT	336
Triazine Resistant	AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	337
D1Protein	CTTCCAATATGCTA C TTTTAACAACTCTCGCTCTTTACATTTCTTCT
Populus deltoides	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAG	338
AGT-ACT	CGAGAGTTGTTAAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATATTATAAGTTT
	ATATGCTA C TTTTAACA	339
	TGTTAAAA G TAGCATAT	340
Triazine Resistant	AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	341
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Petunia x hybrida	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA	342
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	343
	TGTTGAAA G TAGCATAT	344
Triazine Resistant	AAACTTATAAIATCGTAGCTGCTCATGGTTATTTTGGCCGATTGAT	345
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCC
Magnolia pyramidata	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA	346
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCAGCTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	347
	TGTTGAAA G TAGCATAT	348
Triazine Resistant	AAACCTATAATATTGTAGCAGCTCATGGTTATTTTGGCCGATTGAT	349
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCTTTACATTTCTTCC
Medicago sativa	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA	350
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAAGCATGAGCTGCTACAATATTATAGGTTT
	ATATGCTA C TTTCAACA	351
	TGTTGAAA+E,us GTAGCATAT	1352
Triazine Resistant	AAACCTATAATATTGTAGCTGCTCATGGTTATTTGGCCGATTGAT	353
D1Protein	CTTCCAATATGCAA C TTTCAACAATTCTCGTTCTTTACATTTCTTCT
Glycine max	TAGCTGCTTGGCCTGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA	354
AGT-ACT	CGAGAATTGTTGAAA G TTGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCAGCTACAATATTATAGGTTT
	ATATGCAA C TTTCAACA	355
	TGTTGAAA G TTGCATAT	356
Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	357
D1Protein	CTTCCAATATGCT AC TTTCAACAATTCTCGTTCTTTACATTTCTTCT
Brassica napus	TAGCGGCTTGGCCGGTAGTAGGTATTTG
Gly264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	358
GGT-ACT	CGAGAAITGTTGAAA GT AGCATATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCT AC TTTCAACA	359
	TGTTGAAA GT AGCATAT	360
Triazine Resistant	AAACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT	361
D1Protein	CTTCCAATATGCTA C TTTTAACAACTCTCGTTCTTTACACTTCTTCT
Oryza sativa	TGGCTGCTTGGCCTGTAGTAGGGATTTG
Ser264Thr	CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA	362
AGT-ACT	CGAGAGTTGTTAAAA G TAGCATATTGGAAGATTAATCGGCCAAAAT
	AACCATGAGCGGCCACAATATTATAAGTTT
	ATATGCTA C TTTTAACA	363
	TGTTAAAA G TAGCATAT	364
Triazine Resistant	AGACTTATAATATTGTGGCTGCTCACGGTTATTTTGGTCGATTAAT	365
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACACTTCTTCT
Zea mays	TGGCTGCTtGGCCTGTAGTAGGGATCtG
Ser264Thr	CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA	366
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATTAATCGACCAAAAT
	AACCGTGAGCAGCCACAATATTATAAGTCT
	ATATGCTA C TTTCAACA	367
	TGTTGAAAGTAGCATAT	368
Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	369
D1Protein	TTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCTT
Arabidopsis thaliana	AGCGGCTTGGCCGGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	370
AGT-ACT	CGAGAATTGITGAAA G TAGCATATTGGAAAATCAATCGGCCAAAAT
	AACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCTA C TTTCAACA	371
	TGTTGAAA G TAGCATAT	372
Triazine Resistant	AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	373
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Nicotiana tabacum	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA	374
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATGTTATAAGTTT
	ATATGCTA C TTTCAACA	375
	TGTTGAAA GTAGCATAT	376
Triazine Resistant	AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	377
D1Protein	CTTCCAATATGCTA C TTTTAACAACTCTCGCTCTTTACATTTCTTCT
Papulus deltoides	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGGTAAGAAGAAATGTAAAGAG	378
AGT-AGT	CGAGAGTTGTTAAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATATTATAAGTTT
	ATATGCTA C TTTTAACA	379
	TGTTAAAA G TAGCATAT	380
Triazine Resistant	AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	381
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Petunia x hybrida	TAGCTGGTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA	382
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	383
	TGTTGAAA G TAGCATAT	384
Triazine Resistant	AAACTTATAATATCGTAGCTGCTCATGGTTATTTTGGCCGATTGAT	385
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCC
Magnolia pyramidata	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA	386
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCAGCTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	387
	TGTTGAAA G TAGCATAT	388
Triazine Resistant	AAACCTATAATATTGTAGCAGCTCATGGTTATTTTGGCCGATTGAT	389
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCTTTACATTTGTTCC
Medicago sativa	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA	390
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCTGCTACAATATTATAGGTTT
	ATATGCTA C TTTCAACA	391
	TGTTGAAA G TAGCATAT	392
Triazine Resistant	AAACCTATAATATTGTAGCTGCTCATGGTTATTTTGGCCGATTGAT	393
D1Protein	CTTCCAATATGCAA C TTTCAACAATTCTCGTTCTTTACATTTCTTCT
Glycine max	TAGCTGCTTGGCCTGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAATGTAAAGAA	394
AGT-ACT	CGAGAATTGTTGAAA G TTGCATATTGGAAGATCAATCGGGCAAAA
	TAACCATGAGCAGCTACAATATTATAGGTTT
	ATATGCAA C TTTCAACA	395
	TGTTGAAA G TTGCATAT	396
Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	397
D1Protein	CTTCCAATATGCT AC TTTCAACAATTCTCGTTCTTTACATTTCTTCT
Brassica napus	TAGCGGCTTGGCCGGTAGTAGGTATTTG
Gly264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	398
GGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCT AC TTTCAACA	399
	TGTTGAAA GT AGCATAT	400
Triazine Resistant	AAACTTATAATATTGTGGCCGCTCATGGTTATTTTGGCCGATTAAT	401
D1Protein	CTTCCAATATGCTA C TTTTAACAACTCTCGTTCTTTACACTTCTTCT
Oryza sativa	TGGCTGCTTGGCCTGTAGTAGGGATTTG
Ser264Ihr	CAAATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA	402
AGT-ACT	CGAGAGTTGTTAAAA G TAGCATATTGGAAGATTAATCGGCCAAAAT
	AACCATGAGCGGCCACAATATTATAAGTTT
	ATATGCTA C TTTTAACA	403
	TGTTAAAA G TAGCATAT	404
Triazine Resistant	AGACTTATAATATTGTGGCTGCTCACGGTTATTTTGGTCGATTAAT	405
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACACTTCTTCT
Zea mays	TGGCTGCTTGGCCTGTAGTAGGGATCTG
Ser264Thr	CAGATCCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA	406
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATTAATCGACCAAAAT
	AACCGTGAGCAGCCACAATATTATAAGTCT
	ATATGCTA C TTTCAACA	407
	TGTTGAAAGTAGCATAT	408
Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	409
D1Protein	TTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCTT
Arabidopsis thaliana	AGCGGCTTGGCCGGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	410
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAAATCAATCGGCCAAAAT
	AACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCTA C TTTCAACA	411
	TGTTGAAA G TAGCATAT	412
Triazine Resistant	AAACCTACAATATTGTGGCTGCTCACGGTTATTTCGGCCGATTGAT	413
D1Protein	CTTCCAGTATGCTA C TTTCAACAACTCCCGTTCTTTACATTTCTTCT
Picea abies	TAGCTGCTTGGCCCGTAGCAGGTATCTG
Ser264Thr	CAGATACCTGCTACGGGCCAAGCAGCTAAGAAGAAATGTAAAGAA	414
AGT-ACT	CGGGAGTTGTTGAAA G TAGCATACTGGAAGATCAATCGGCCGAAA
	TAACCGTGAGCAGCCACAATATTGTAGGTTT
	GTATGCTA C TTTCAACA	415
	TGTTGAAA G TAGCATAC	416
Triazine Resistant	AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	417
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGCTCTTTACATTTCTTCC
Vicia faba	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAG	418
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATATTATAGGTTT
	ATATGCTA C TTTCAACA	419
	TGTTGAAA G TAGCATAT	420
Triazine Resistant	AGACTTATAATATTGTGGCTGCTCATGGTTATTTTGGCCGATTAAT	421
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCTTTACACTTCTTCT
Hordeum vulgare	TGGCTGCTTGGCCTGTAGTAGGAATCTG
Ser264Thr	CAGATTCCTACTACAGGCCAAGCAGCCAAGAAGAAGTGTAAAGAA	422
AGT-ACT	CGAGAGTTGTTGAAAGTAGCATATTGGAAGATTAATCGGCCAAAA
	TAACCATGAGCAGCCACAATATTATAAGTCT
	ATATGCTACTTTCAACA	423
	TGTTGAAAGTAGCATAT	424
Triazine Resistant	AAACTTATAATATTGTGGCTGCTCATGGTTATTTTGGCCGATTAAT	425
D1Protein	CTTCCAATATGCTACTTTCAACAACTCTCGTTCTTTACACTTCTTCT
Triticum aestivum	TGGCTGCTTGGCCTGTAGTAGGAATCTG
Ser264Thr	CAGATTCCTACTACAGGCCMGCAGCCAAGAAGAAGTGTAAAGAA	426
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATTAATCGGCCAAAA
	TAACCATGAGCAGCCACAATATTATAAGTTT
	ATATGCTA C TTTCAACA	427
	TGTTGAAA G+E TAGCATAT	428
Triazine Resistant	AAACTTATAATATTGTAGCTGCTCATGGTTATTTTGGCCGATTAATC	429
D1Protein	TTCCAATATGCAA C TTTCMCAATTCTCGTTCTTTACATTTCTTCCT
Vigna unguiculata	AGCTGCTTGGCCTGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACAGGCCAAGCAGCTAGGAAGAAATGTAAAGAA	430
AGT-ACT	CGAGAATTGTTGAAA G TTGCATATTGGAAGATTAATCGGCCAAAAT
	AACCATGAGCAGCTACAATATTATAAGTTT
	ATATGCAA C TTTCAACA	431
	TGTTGAAA G TTGCATAT	432
Triazine Resistant	AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	433
D1Protein	CTTCCAATATGCAA C TTTCAACAACTCTCGTTCTTTACACTTCTTCT
Lotus japonicus	TAGCTGCTTGGCCTGTTGTAGGTATCTG
Ser264Thr	CAGATACCTACAACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA	434
AGT-ACT	CGAGAGTTGTTGAAA G TTGCATATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATATTATAGGTTT
	ATATGCAA C TTTCAACA	435
	TGTTGAAA G TTGCATAT	436
Triazine Resistant	AAACTTACAACATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	437
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGTTCTTTACATTTCTTCT
Sinapis alba	TAGCGGCTTGGCCGGTAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACCGGCCAAGCCGCTAAGAAGAAATGTAAAGAA	438
AGT-ACT	CGAGAATTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATGTTGTAAGTTT
	ATATGCTA C TTTCAACA	439
	TGTTGAAA G TAGCATAT	440
Triazine Resistant	AAACCTATAATATTGTAGCTGCTCACGGTTATTTTGGCCGATTGAT	441
D1Protein	CTTCCAATATGCTA C TTTCAACAATTCTCGCTCTTTACATTTCTTCC
Pisum sativum	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTIAGGAAGAAATGTAAAGAG	442
AGt-ACT	CGAGAATTGTTGAAAGTAGCAtATTGGAAGATCAATCGGCCAAAA
	TAACCGTGAGCAGCTACAATATTATAGGTTT
	ATATGCTA C TTTCAACA	443
	TGTTGAAA G TAGCATAT	444
Triazine Resistant	AAACTTATAATATCGTAGGTGCTCATGGTTATTTTGGTCGATTGAT	445
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCTTTACACTTCTTCT
Spinacia oleracea	TAGCTGCTTGGCCTGIAGTAGGTATTTG
Ser264Thr	CAAATACCTACTACAGGCCAAGCAGCTAAGAAGAAGTGTAAAGAA	446
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGACCAAAA
	TAACCATGAGCAGGTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	447
	TGTTGAAA G TAGCATAT	448
Triazine Resistant	AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	449
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Nicotiana debneyi	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGAtACCtACTACAGGCGAAGCAGCtAGGAAGAAGTGTAACGAA	450
AGT-ACT	CGAGAGtTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATGTTATAAGTTT
	ATATGCTA C TTTCAACA	451
	TGTTGAAA G TAGCATAT	452
Triazine Resistant	AAACTTATAATATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	453
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTTCGTTACACTTCTTCC
Solanum nigrum	TAGCTGCTTGGCCTGTAGTAGGTATCTG
Ser264Thr	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA	454
AGT-ACT	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
	TAACCATGAGCGGCTACGATATTATAAGTTT
	ATATGCTA C TTTCAACA	455
	TGTTGAAA G TAGCATAT	456
Triazine Resistant	AAACTTATAACATCGTAGCCGCTCATGGTTATTTTGGCCGATTGAT	457
D1Protein	CTTCCAATATGCTA C TTTCAACAACTCTCGTICGTTACACTTCTTCC
Nicotiana	TAGCTGCTTGGCCTGTAGTAGGTATCTG
plumbaginifolia	CAGATACCTACTACAGGCCAAGCAGCTAGGAAGAAGTGTAACGAA	458
Ser264Thr	CGAGAGTTGTTGAAA G TAGCATATTGGAAGATCAATCGGCCAAAA
AGT-ACT	TAACCATGAGCGGCTACGATGTTATAAGTTT
	ATATGCTA C TTTCAACA	459
	TGTTGAAA G TAGCATAT	460

EXAMPLE 6

Engineering Male- or Female-Sterile Plants

Flower development in distantly related dicot plant species is increasingly better understood and appears to be regulated by a family of genes which encode regulatory proteins. These genes include, for example, AGAMOUS (AG), APETALA1 (AP1), and APETALA3 (AP3) and PISTILLATA (PI) in [0123] Arabidopsis thaliana, and DEFICIENS A (DEFA), GLOBOSA (GLO), SQUAMOSA (SQUA), and PLENA (PLE) in Antirrhinum majus. Genetic studies have shown that the DEFA, GLO and AP3 genes are essential for petal and stamen development. Sequence analysis of these genes revealed that the gene products contain a conserved MADS box region, a DNA-binding domain. Using these clones as probes, MADS box genes have also been isolated from other species including tomato, tobacco, petunia, Brassica napus, and maize.
Altering the expression of these genes results in altered floral morphology. For example, mutations in AP3 and PI result in male-sterile flowers because petals develop in place of stamens. [0124]

The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer altered floral structures in plants.

TABLE 14


Oligonucleotides to produce male-sterile plants

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Male-sterile	TTGTCCTCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAG	461
AP3	AGAAGAATATGGCG T GAGGGAAGATCCAGATCAAGAGGATAGAGA
Arabidopsis thaliana	ACCAGACAAACAGACAAGTGACGTATTCAA
Arg3Term	TTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTCTTGATC	462
AGA-TGA	TGGATCTTCCCTC A CGCCATATTCTTCTCTCTTIGTTTAATCTTTTT
	GTTGAAGAGATTTGGTGGAGAGGACAA
	ATATGGCG T GAGGGAAG	463
	CTTCCCTC A CGCCATAT	464

Male-sterile	TCTCCACCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGA	465
AP3	ATATGGCGAGAGGG T AGATCCAGATCAAGAGGATAGAGAACCAGA
Arabidopsis thaliana	CAAACAGACAAGTGACGTATTCAAAGAGAA
Lys5Term	TTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCTATCCTC	466
AAG-TAG	TTGATCTGGATCT A CCCTCTCGCCATATTCTTCTCTCTTTGTTTAAT
	CTTTTTGTTGAAGAGATTTGGTGGAGA
	CGAGAGGG T AGATCCAG	467
	CTGGATCT A CCGTCTCG	468

Male-sterile	CCAAATCTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGG	469
AP3	CGAGAGGGAAGATC T AGATCAAGAGGATAGAGAAGCAGACAAACA
Arabidopsis thaliana	GACAAGTGACGTATTCAAAGAGAAGGAATG
Gln7Term	CATTCCTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGGTTCTCT	470
CAG-TAG	ATCCTCTTGATCT A GATCTTCCCTCTCGCCATATTCTTCTCTCTTTG
	TTTAATCTTTTTGTTGAAGAGATTTGG
	GGAAGATC T AGATCAAG	471
	CTTGATCT A GATCTTCC	472

Male-sterile	CTCTTCAACAAAAAGATTAAACAAAGAGAGAAGAATATGGCGAGAG	473
AP3	GGAAGATCCAGATC T AGAGGATAGAGAACCAGACAAACAGAGAAG
Arabidopsis thaliana	TGACGTATTCAAAGAGAAGGAATGGTTTAT
Lys9Term	ATAAACCATTCGTTCTCTTTGAATACGTCACTTGTCTGTTTGTCTGG	474
AAG-TAG	TTCTCTATCCTCT A GATCTGGATCTTCCCTCTCGCCATATTCTTCTC
	TCTTTGTTTAATCTTTTTGTTGAAGAG
	TCCAGATC T AGAGGATA	475
	TATCCTCT A GATCTGGA	476

Male-sterile	AGAGGGAAGATCGAGATGAAGAGGATAGAGAACGAGAGGAACCG	477
AP3	ACAAGTGACGTATTCT T AGAGAAGAAATGGTTTGTTCAAGAAAGCT
Brassica oleracea	CACGAGCTTACAGTTTTATGTGATGCTAGGG
Lys23Term	CCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAACAA	478
AAG-TAG	ACCATTTCTTCTCT A AGAATACGTCACTTGTCGGTTGGTCTGGTTC
	TCTATCCTCTTGATCTGGATCTTCCCTCT
	CGTATTCT T AGAGAAGA	479
	TCTTCTCT A AGAATACG	480

Male-sterile	GGGAAGATCCAGATCAAGAGGATAGAGAACCAGACCAACCGACAA	481
AP3	GTGACGTATTCTAAG T GAAGAAATGGTTTGTTCAAGAAAGCTCACG
Brassica oleracea	AGCTTACAGTTTTATGTGATGCTAGGGTTT
Arg24Term	AAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTTGAA	482
AGA-TGA	CAAACCATTTCTTC A CTTAGAATACGTCACTTGTGGGTTGGTCTGG
	TTCTCTATCCTCTTGATCTGGATCTTCCC
	ATTCTAAG T GAAGAAAT	483
	ATTTCTTC A CTTAGAAT	484

Male-sterile	AAGATCCAGATCAAGAGGATAGAGAACCAGACCAACCGACAAGTG	485
AP3	ACGTATTCTAAGAGA T GAAATGGTTTGTTCAAGAAAGCTCACGAGC
Brassica oleracea	TTACAGTTTTATGTGATGCTAGGGTTTCGA
Arg25Term	TCGAAACCCTAGCATCACATAAAACTGTAAGCTCGTGAGCTTTCTT	486
AGA-TGA	GAACAAACCATTTC A TCTCTTAGAATACGTCACTTGTCGGTTGGTC
	TGGTTCTCTATGCTCTTGATCTGGATCTT
	CTAAGAGA T GAAATGGT	487
	ACCATTTC A TCTCTTAG	488

Male-sterile	TCAAGAGGATAGAGAACCAGACCAACCGACAAGTGACGTATTCTA	489
AP3	AGAGAAGAAATGGTT A GTTCAAGAAAGCTCACGAGCTTACAGTTTT
Brassica oleracea	ATGTGATGCTAGGGTTTCGATTATCATGTT
Leu28Term	AACATGATAATCGAAACCCTAGCATCACATAAAACTGTAAGCTCGT	490
TTG-TAG	GAGCTTTCTTGAAC T AACCATTTCTTCTCTTAGAATACGTCACTTGT
	CGGTTGGTCTGGTTCTCTATCCTCTTGA
	AAATGGTT A GTTCAAGA	491
	TCTTGAAC T AACCATTT	492

Male-sterile	GGCTCGAGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAA	493
AP3	CAGGCAGGTCACCTA G TCCAAGAGAAGAAATGGTTTGTTCAAGAA
Brassica napus	AGCACACGAGCTCTCTGTTCTCTGTGATGCT
Tyr21Term	AGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAAACC	494
TAC-TAG	ATTTCTTCTCTTGGA C TAGGTGACCTGCCTGTTTGTTTGGTTCTCTA
	TCCTCTTAATCTGGATCTTCCCTCGAGCC
	GTCACCTA G TCCAAGAG	495
	CTCTTGGA C TAGGTGAC	496

Male-sterile	CGAGGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGG	497
AP3	CAGGTCACCTACTCC T AGAGAAGAAATGGTTTGTTCAAGAAAGCAC
Brassica napus	ACGAGCTCTCTGTTCTCTGTGATGCTAAAG
Lys23Term	CTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAACAA	498
AAG-TAG	ACCATTTCTTCTCT A GGAGTAGGTGACCTGCCTGTTTGTTTGGTTC
	TCTATCCTCTTAATCTGGATCTTCCCTCG
	CCTACTCC T AGAGAAGA	499
	TCTTCTCT A GGAGTAGG	500

Male-sterile	GGGAAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAG	501
AP3	GTCACCTACTCCAAG T GAAGAAATGGTTTGTTCAAGAAAGCACACG
Brassica napus	AGCTCTCTGTTCTCTGTGATGCTAAAGTTT
Arg24Term	AAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTTGAA	502
AGA-TGA	CAAACCATTTGTTC A CTTGGAGTAGGTGACCTGCCTGTTTGTTTGG
	TTCTCTATCCTCTTAATCTGGATCTTCCC
	ACTCCAAG T GAAGAAAT	503
	ATTTCTTC A CTTGGAGT	504

Male-sterile	AAGATCCAGATTAAGAGGATAGAGAACCAAACAAACAGGCAGGTC	505
AP3	ACCTACTCCAAGAGA T GAAATGGTTTGTTCAAGAAAGCACACGAG
Brassica napus	CTCTCTGTTCTCTGTGATGCTAAAGTTTCCA
Arg25Term	TGGAAACTTTAGCATCACAGAGAACAGAGAGCTCGTGTGCTTTCTT	506
AGA-TGA	GAACAAACCATTTC A TCTCTTGGAGTAGGTGACCTGCCTGTTTGTT
	TGGTTCTCTATCCTCTTAATCTGGATCTT
	CCAAGAGA T GAAATGGT	507
	ACCATTTC A TCTCTTGG	508

Male-sterile	GGAGAGAAAGGAAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGG	509
DEFA	TAGTGGTTCGATGGCT T GAGGGAAGATCCAGATTAAGAGGATAGA
Antirrhinum majus	GAACCAAACAAACAGGCAGGTCACCTACTCCA
Arg3Term	TGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCTCTTAAT	510
CGA-TGA	CTGGATCTTCCCTC A AGCCATCGAACCACTACCACTACTGCTCTTG
	TTTTCTTCTTCCAGCTTTCCTTTCTCTCC
	CGATGGCT T GAGGGAAG	511
	CTTCCCTC A AGCCATCG	512

Male-sterile	AAAGGAAAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGT	513
DEFA	TCCATGGCTCGAGGG T AGATCCAGATTAAGAGGATAGAGAACCAA
Antirrhinum majus	ACAAACAGGCAGGTCACCTACTCCAAGAGAA
Lys5Term	TTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTCTATCCT	514
AAG-TAG	CTTAATCTGGATCT A CCCTCGAGCCATCGAACCACTAGCACTACTG
	CTCTTGTTTTCTTCTTCCAGCTTTCCTTT
	CTCGAGGG T AGATCCAG	515
	CTGGATCT A CCCTCGAG	516

Male-sterile	AAGCTGGAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATG	517
DEFA	GCTCGAGGGAAGATC T AGATTAAGAGGATAGAGAACCAAACAAAC
Antirrhinum majus	AGGCAGGTCACCTACTCCAAGAGAAGAAATG
Gln7Term	CATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTGGTTCTC	518
CAG-TAG	TATCCTCTTAATCT A GATCTTCCCTCGAGCCATCGAACCACTACCA
	CTACTGCTCTTGTTTTCTTCTTCCAGCTT
	GGAAGATC T AGATTAAG	519
	CTTAATCT A GATCTTCC	520

Male-sterile	GAAGAAGAAAACAAGAGCAGTAGTGGTAGTGGTTCGATGGCTCGA	521
DEFA	GGGAAGATCCAGATT T AGAGGATAGAGAACCAAACAAACAGGCAG
Antirrhinum majus	GTCACCTACTCCAAGAGAAGAAATGGTTTGT
Lys9Term	ACAAACCATTTCTTCTCTTGGAGTAGGTGACCTGCCTGTTTGTTTG	522
AAG-TAG	GTTCTCTATCCTCT A AATCTGGATCTTCCCTCGAGCCATCGAACCA
	CTACCACTACTGCTCTTGTTTTCTTCTTC
	TCCAGATT T AGAGGATA	523
	TATCCTCT A AATCTGGA	524

Male-sterile	TCAGTAATTCTTAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAAC	525
AP3	TATGGCTCGTGGG T AGATCCAGATCAAGAGAATAGAGAACCAAAC
Nicotiana tabacum	AAACAGACAAGTCACTTATTCTAAGAGAA
Lys5Term	TTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCTATTCTC	526
AAG-TAG	TTGATCTGGATCT A CCCACGAGCCATAGTTTTTTTTTCTTTTTGCTC
	AAAGTTTGAGATCTTAAGAATTACTGA
	CTCGTGGG T AGATCCAG	527
	CTGGATCT A CCCACGAG	528

Male-sterile	ATTCTTAAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGC	529
AP3	TCGTGGGAAGATC T AGATCAAGAGAATAGAGAACCAAACAAACAG
Nicotiana tabacum	ACAAGTCACTTATTCTAAGAGAAGAAATG
Gln7Term	CATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGGTTCTCT	530
CAG-TAG	ATTCTCTTGATCT A GATCTTCCCACGAGCCATAGTTTTTTTTTCTTT
	TTGCTCAAAGTTTGAGATCTTAAGAAT
	GGAAGATC T AGATCAAG	531
	CTTGATCT A GATCTTCC	532

Male-sterile	AAGATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTG	533
AP3	GGAAGATCCAGATC T AGAGAATAGAGAACCAAACAAACAGACAAG
Nicotiana tabacum	TCACTTATTCTAAGAGAAGAAATGGACTTT
Lys9Term	AAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTTTGG	534
AAG-TAG	TTCTCTATTCTCT A GATCTGGATCTTCCCACGAGCCATAGTTTTTTT
	TTCTTTTTGCTCAAAGTTTGAGATCTT
	TCCAGATC T AGAGAATA	535
	TATTCTCT+E,un AGATCTGGA	536

Male-sterile	ATCTCAAACTTTGAGCAAAAAGAAAAAAAAACTATGGCTCGTGGGA	537
AP3	AGATCCAGATCAAG T GAATAGAGAACCAAACAAACAGACAAGTCA
Nicotiana tabacum	CTTATTCTAAGAGAAGAAATGGACTTTTCA
Arg10Term	TGAAAAGTCCATTTCTTCTCTTAGAATAAGTGACTTGTCTGTTTGTT	538
AGA-TGA	TGGTTCTCTATTC A CTTGATCTGGATCTTCCCACGAGCCATAGTTT
	TTTTTTCTTTTTGCTCAAAGTTTGAGAT
	AGATCAAG T GAATAGAG	539
	CTCTATTC A CTTGATCT	540

Male-sterile	GGCTCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAA	541
AP3	CAGACAAGTAACTTA G TCAAAACGAAGGGATGGTCTTTTCAAGAAG
Medicago sativa	GCCAATGAGCTCACTGTTCTTTGTGATGCT
Tyr21Term	AGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGACCA	542
TAC-TAG	TCCCTTCGTTTTGA C TAAGTTACTTGTCTGTTCGTTGTGTTCTCTAT
	TCTCTTGATCTGGATCTTTCCTCGAGCC
	GTAACTTA G TCAAAACG	543
	CGTTTTGA C TAAGTTAC	544

Male-sterile	CTCGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACA	545
AP3	GACAAGTAACTTACT G AAAACGAAGGGATGGTCTTTTCAAGAAGG
Medicago sativa	CCAATGAGCTCACTGTTCTTTGTGATGCTAA
Ser22Term	TTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAGAC	546
TCA-TGA	CATCCCTTCGTTTT C AGTAAGTTACTTGTCTGTTCGTTGTGTTCTCT
	ATTCTCTTGATCTGGATCTTTCCTCGAG
	AACTTACT G AAAACGAA	547
	TTCGTTTT+E,un CAGTAAGTT	548

Male-sterile	CGAGGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACAGA	549
AP3	CAAGTAACTTACTCA T AACGAAGGGATGGTCTTTTCAAGAAGGCCA
Medicago sativa	ATGAGCTCACTGTTCTTTGTGATGCTAAGG
Lys23Term	CCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAAAAG	550
AAA-TAA	ACCATCCCTTCGTT A TGAGTAAGTTACTTGTCTGTTCGTTGTGTTCT
	CTATTCTCTTGATCTGGATCTTTCCTCG
	CTTACTCA T AACGAAGG	551
	CCTTCGTT A TGAGTAAG	552

Male-sterile	GGAAAGATCCAGATCAAGAGAATAGAGAACACAACGAACAGACAA	553
AP3	GTAACTTACTCAAAA T GAAGGGATGGTCTTTTCAAGAAGGCCAATG
Medicago sativa	AGCTCACTGTTCTTTGTGATGCTAAGGTTT
Arg24Term	AAACCTTAGCATCACAAAGAACAGTGAGCTCATTGGCCTTCTTGAA	554
CGA-TGA	AAGACCATCCCTTC A TTTTGAGTAAGTTACTTGTCTGTTCGTTGTGT
	TCTCTATTCTCTTGATCTGGATCTTTCC
	ACTCAAAA T GAAGGGAT	555
	ATCCCTTC A TTTTGAGT	556

Male-sterile	GGCTCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAAT	557
DEF4	AGGCAAGTGACTTA G TCAAAGAGAAGAAATGGGCTATTCAAGAAG
Solanum tuberosum	GCTAATGAACTTACAGTTCTTTGTGATGCT
Tyr21Term	AGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCCCA	558
TAT-TAG	TTTCTTCTCTTTGA C TAAGTCACTTGCCTATTTGTTTGGTTTTCTATT
	TTCTTGATCTGGATCTTACCACGAGCC
	GTGACTTA G TCAAAGAG	559
	CTCTTTGA C TAAGTCAC	560

Male-sterile	CTCGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAG	561
DEF4	GCAAGTGACTTATT G AAAGAGAAGAAATGGGCTATTCAAGAAGGC
Solanum tuberosum	TAATGAACTTACAGTTCTTTGTGATGCTAA
Ser22Term	TTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAGCC	562
TCA-TGA	CATTTCTTCTCTTT C AATAAGTCACTTGCCTATTTGTTTGGTTTTCTA
	TTTTCTTGATCTGGATCTTACCACGAG
	GACTTATT G AAAGAGAA	563
	TTCTCTTT C AATAAGTC	564

Male-sterile	CGTGGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGG	565
DEF4	CAAGTGACTTATTCA T AGAGAAGAAATGGGCTATTCAAGAAGGCTA
Solanum tuberosum	ATGAACTTACAGTTCTTTGTGATGCTAAAG
Lys23Term	CTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAATAG	566
AAG-TAG	CCCATTTCTTCTCT A TGAATAAGTCACTTGCCTATTTGTTTGGTTTT
	CTATTTTCTTGATCTGGATCTTACCACG
	CTTATTCA T AGAGAAGA	567
	TCTTCTCT A TGAATAAG	568

Male-sterile	GGTAAGATCCAGATCAAGAAAATAGAAAACCAAACAAATAGGCAA	569
DEF4	GTGACTTATTCAAAG T GAAGAAATGGGCTATTCAAGAAGGCTAATG
Solanum tuberosum	AACTTACAGTTCTTTGTGATGCTAAAGTTT
Arg24Term	AAACTTTAGCATCACAAAGAACTGTAAGTTCATTAGCCTTCTTGAAT	570
AGA-TGA	AGCCCATTTCTTC A CTTTGAATAAGTCACTTGCCTATTTGTTTGGTT
	TTCTATTTTCTTGATCTGGATCTTACC
	ATTCAAAG T GAAGAAAT	571
	ATTTCTTC A GTTTGAAT	572

Male-sterile	GCTAATGAACTTACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTAT	573
AP3	GATTTCTAGTACT T GAAAACTTCATGAGTTTATAAGTCCCTCTATCA
Lycopersicon	CGACCAAACAATTGTTCGATCTGTACC
esculentum	GGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTATAAA	574
Gly27Term	CTCATGAAGTTTTC A AGTACTAGAAATCATAACAATTGAAACTTTAG
GGA-TGA	CATCACAAAGAACAGTAAGTTCATTAGC
	CTAGTACT T GAAAACTT	575
	AAGTTTTC A AGTACTAG	576

Male-sterile	AATGAACTTACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGAT	577
AP3	TTCTAGTACTGGA T AACTTCATGAGTTTATAAGTCCCTCTATCACGA
Lycopersicon	CCAAACAATTGTTCGATCTGTACCAGA
esculentum	TCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGAGGGACTTAT	578
Lys28Term	AAACTCATGAAGTT A TCCAGTACTAGAAATCATAACAATTGAAACTT
AAA-TAA	TAGCATCACAAAGAACAGTAAGTTCATT
	GTACTGGA T AACTTCAT	579
	ATGAAGTT A TCCAGTAC	580

Male-sterile	ACTGTTCTTTGTGATGCTAAAGTTTCAATTGTTATGATTTCTAGTAC	581
AP3	TGGAAAACTTCAT T AGTTTATAAGTCCCTCTATCACGACCAAACAAT
Lycopersicon	TGTTCGATCTGTACCAGAAGACTATTG
esculentum	CAATAGTCTTCTGGTACAGATCGAACAATTGTTTGGTCGTGATAGA	582
Glu31Term	GGGACTTATAAACT A ATGAAGTTTTCCAGTACTAGAAATCATAACA
GAG-TAG	ATTGAAACTTTAGCATCACAAAGAACAGT
	AACTTCAT T AGTTTATA	583
	TATAAACT A ATGAAGTT	584

Male-sterile	ATTGTTATGATTTCTAGTACTGGAAAACTTCATGAGTTTATAAGTCC	585
AP3	CTCTATCACGACC T AACAATTGTTCGATCTGTACCAGAAGACTATT
Lycopersicon	GGAGTTGATATTTGGACTACTCACTATG
esculentum	CATAGTGAGTAGTCCAAATATCAACTCCAATAGTCTTCTGGTACAG	586
Lys40Term	ATCGAACAATTGTT A GGTCGTGATAGAGGGACTTATAAACTCATGA
AAA-TAA	AGTTTTCCAGTACTAGAAATCATAACAAT
	TCACGACC T AACAATTG	587
	CAATTGTT A GGTCGTGA	588

Male-sterile	GGGGCGGGGGAAGATTGAGATAAAGCGGATCGAGAACGCCACCA	589
AP3	ACAGGCAGGTGACCTA G TCCAAGCGCCGGTCGGGGATCATGAAG
Triticum aestivum	AAGGCGCGGGAGCTCACCGTGCTCTGCGACGCC
Tyr21Term	GGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATGATCC	590
TAC-TAG	CCGACCGGCGCTTGGA C TAGGTCACCTGCCTGTTGGTGGCGTTCT
	CGATCCGCTTTATCTCAATCTTCCCCCGCCCC
	GTGACCTA G TCCAAGCG	591
	CGCTTGGA C TAGGTCAC	592

Male-sterile	CGGGGGAAGATTGAGATAAAGCGGATCGAGAACGCCACCAACAG	593
AP3	GCAGGTGACCTACTCC T AGCGCCGGTCGGGGATCATGAAGAAGG
Triticum aestivum	CGCGGGAGCTCACCGTGCTCTGCGACGCCCAGG
Lys23Term	CCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGCCTTCTTCATG	594
AAG-TAG	ATCCCCGACCGGCGCT A GGAGTAGGTCACCTGCCTGTTGGTGGC
	GTTCTCGATCCGCTTTATCTCAATCTTCCCCCG
	CCTACTCC T AGCGCCGG	595
	CCGGCGCT A GGAGTAGG	596

Male-sterile	TTGAGATAAAGCGGATCGAGAACGCCACCAACAGGCAGGTGACCT	597
AP3	ACTCGAAGCGCCGGT A GGGGATCATGAAGAAGGCGCGGGAGCTC
Triticum aestivum	ACCGTGCTCTGCGACGCCCAGGTCGCCATCAT
Ser26Term	ATGATGGCGACCTGGGCGTCGCAGAGCACGGTGAGCTCCCGCGC	598
TCG-TAG	CTTCTTCATGATCCCC T ACCGGCGCTTGGAGTAGGTCACCTGCCT
	GTTGGTGGCGTTGTCGATCCGCTTTATCTCAA
	GCGCCGGT A GGGGATCA	599
	TGATCCCC T ACCGGCGC	600

Male-sterile	CGGATCGAGAACGCCACCAACAGGCAGGTGACCTACTCCAAGCG	601
AP3	CCGGTCGGGGATCATG T AGAAGGCGCGGGAGCTCACCGTGCTCT
Triticum aestivum	GCGACGCCCAGGTCGCCATCATCATGTTCTCCT
Lys30Term	AGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG	602
AAG-TAG	AGCTCCCGCGCCTTCT A CATGATCCCCGACCGGCGCTTGGAGTAG
	GTCACCTGCCTGTTGGTGGCGTTGTCGATCCG
	GGATCATG T AGAAGGCG	603
	CGCCTTCT A CATGATCC	604

Male-sterile	GGGGCGCGGCAAGATCGAGATCAAGCGGATCGAGAACGCCACCA	605
Silky1	ACCGCCAGGTGACCTA G TCCAAGCGCCGGACGGGGATCATGAAG
Zea mays	AAGGCACGCGAGCTCACCGTGCTCTGCGACGCC
Tyr21Term	GGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATGATCCC	606
TAG-TAG	CGTCCGGCGCTTGGA C TAGGTCACCTGGCGGTTGGTGGCGTTCT
	CGATCGGCTTGATCTCGATCTTGCCGCGCCCC
	GTGACCTA G TCCAAGCG	607
	CGCTTGGA C TAGGTCAC	608

Male-sterile	CGCGGCAAGATCGAGATCAAGCGGATCGAGAACGCCACCAACCG	609
Silky1	CCAGGTGACCTACTCC T AGCGCCGGACGGGGATCATGAAGAAGG
Zea mays	CACGCGAGCTCACCGTGCTCTGCGACGCCCAGG
Lys23Term	CCTGGGCGTCGCAGAGCACGGTGAGCTCGCGTGCCTTCTTCATG	610
AAG-TAG	ATCCCCGTCCGGCGCT A GGAGTAGGTCACCTGGCGGTTGGTGGC
	GTTCTCGATCCGCTTGATCTCGATCTTGCCGCG
	CCTACTCC T AGCGCCGG	611
	CCGGCGCT A GGAGTAGG	612

Male-sterile	CGGATCGAGAACGCCACCAACCGCCAGGTGACCTACTCCAAGCG	613
Silky1	CCGGACGGGGATCATG T AGAAGGCACGCGAGCTCACCGTGCTCT
Zea mays	GCGACGCCCAGGTCGCCATCATCATGTTCTCCT
Lys30Term	AGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACGGTG	614
AAG-TAG	AGCTCGCGTGCCTTCT A CATGATCCCGGTCCGGCGCTTGGAGTAG
	GTCACCTGGCGGTTGGTGGCGTTCTCGATCCG
	GGATCATG T AGAAGGCA	615
	TGCCTTCT A CATGATCC	616

Male-sterile	ATCGAGAACGCCACCAACCGCCAGGTGACGTACTCCAAGCGCCG	617
Silky1	GACGGGGATCATGAAG T AGGCACGCGAGCTCACCGTGCTCTGCG
Zea mays	ACGCCCAGGTCGCCATCATCATGTTCTCCTCCA
Lys31Term	TGGAGGAGAACATGATGATGGCGACCTGGGCGTCGCAGAGCACG	618
AAG-TAG	GTGAGCTCGCGTGCCT A CTTCATGATCCCCGTCCGGCGCTTGGAG
	TAGGTCACCTGGCGGTTGGTGGCGTTCTCGAT
	TCATGAAG T AGGCACGC	619
	GCGTGCCT A CTTCATGA	620

Male-sterile	GCTAGCTGCATTGTCCGGCGAGAGAGATAGCTGCTGCAGGGGGC	621
AP3	GGCCATGGGGAGGGGC T AGATCGAGATCAAGCGGATCGAGAACG
Oryza sativa	CGACCAACAGGCAGGTGACCTACTCGAAGCGCC
Lys5Term	GGCGCTTGGAGTAGGTCACCTGCCTGTTGGTCGCGTTCTCGATCC	622
AAG-TAG	GCTTGATCTCGATCT A GCCCGTCCCCATGGCGGCCCCCTGCAGCA
	GCTATCTCTCTCGCCGGACAATGCAGCTAGC
	GGAGGGGC T AGATCGAG	623
	CTCGATCT A GCCCCTCC	624

Male-sterile	TGCATTGTCCGGCGAGAGAGATAGCTGCTGCAGGGGGCGGCCAT	625
AP3	GGGGAGGGGCAAGATC T AGATCAAGCGGATCGAGAACGCGACCA
Oryza sativa	ACAGGCAGGTGACCTACTCGAAGCGCCGCACGG
Glu7Term	CCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCGCGTTCT	626
GAG-TAG	CGATCCGCTTGATCT A GATCTTGCCCCTCCCCATGGCCGCCCCCT
	GCAGCAGCTATCTCTCTCGCCGGACAATGCA
	GCAAGATC T AGATCAAG	627
	CTTGATCT A GATCTTGC	628

Male-sterile	GTCCGGCGAGAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGA	629
AP3	GGGGCAAGATCGAGATC T AGCGGATCGAGAACGCGACCAACAGG
Oryza sativa	CAGGTGACCTACTCGAAGCGCCGCACGGGGATCA
Lys9Term	TGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCCTGTTGGTCG	630
AAG-TAG	CGTTCTCGATCCGCT A GATCTCGATCTTGCCCCTCCCCATGGCCG
	CCCCCTGCAGCAGCTATCTCTCTCGCCGGAC
	TCGAGATC T AGCGGATC	631
	GATCCGCT A GATCTCGA	632

Male-sterile	GAGAGATAGCTGCTGCAGGGGGCGGCCATGGGGAGGGGCAAGA	633
AP3	TCGAGATCAAGCGGATCT A GAACGCGACCAACAGGCAGGTGACCT
Oryza sativa	ACTCGAAGCGCCGCACGGGGATCATGAAGAAGG
Glu12Term	CCTTCTTCATGATCCCCGTGCGGCGCTTCGAGTAGGTCACCTGCC	634
GAG-TAG	TGTTGGTCGCGTTCT A GATCCGCTTGATCTCGATCTTGCCCCTCCC
	CATGGCGGCCCCCTGCAGCAGCTATCTCTC
	AGCGGATC T AGAACGCG	635
	CGCGTTCT A GATCCGCT	636

TABLE 15


Oligonucleotides to produce male-sterile plants

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Male-sterile	TCTGTACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCA	637
AG	GCAATCACGGCGTA G CAATCGGAGCTAGGAGGAGATTCCTCTCC
Arabidopsis thaliana	CTTGAGGAAATCTGGGAGAGGAAAGATCGAA
Tyr35Term	TTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCCT	638
TAG-TAG	CCTAGGTCCGATTG C TACGCCGTGATTGCTGCTCCAAAGCCAAAA
	ACGTTTAGGGCAAAATTTGATTAGTACAGA
	ACGGCGTA G CAATCGGA	639
	TCCGATTG C TACGCCGT	640

Male-sterile	CTGTACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAG	641
AG	CAATCACGGCGTAC T AATCGGAGCTAGGAGGAGATTCCTCTCCCT
Arabidopsis thaliana	TGAGGAAATCTGGGAGAGGAAAGATCGAAA
Gln36Term	TTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAATCTCC	642
CAA-TAA	TCCTAGCTCCGATT A GTACGCCGTGATTGCTGCTCCAAAGCCAAA
	AACGTTTAGGGCAAAATTTGATTAGTACAG
	CGGCGTAC T AATCGGAG	643
	CTCCGATT A GTACGCCG	644

Male-sterile	ACTAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAGCAAT	645
AG	CACGGCGTACCAAT A GGAGCTAGGAGGAGATTCCTCTCCCTTGA
Arabidopsis thaliana	GGAAATCTGGGAGAGGAAAGATCGAAATCAA
Ser37Term	TTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGAAT	646
TCG-TAG	CTCCTCCTAGCTCC T ATTGGTACGCCGTGATTGCTGCTCCAAAGC
	CAAAAACGTTTAGGGCAAAATTTGATTAGT
	GTACCAAT A GGAGCTAG	647
	CTAGCTCC T ATTGGTAC	648

Male-sterile	TAATCAAATTTTGCCCTAAACGTTTTTGGCTTTGGAGCAGCAATCA	649
AG	CGGCGTACCAATCG T AGCTAGGAGGAGATTCCTCTCCCTTGAGGA
Arabidopsis thalana	AATCTGGGAGAGGAAAGATCGAAATCAAAC
Glu38Term	GTTTGATTTCGATCTTTCCTCTCCCAGATTTCCTCAAGGGAGAGGA	650
GAG-TAG	ATCTCCTCCTAGCT A CGATTGGTACGCCGTGATTGCTGCTCCAAA
	GCCAAAAACGTTTAGGGCAAAATTTGATTA
	ACCAATCG T AGCTAGGA	651
	TCCTAGCT A CGATTGGT	652

Male-sterile	CTCTCCCACTTCTTTTCGGTGGTTTATTCATTTGGTGACGATATCA	653
AG	CAGAAGCAATGGAT T AAGGTGGGAGTAGTCACGATGCAGAGAGT
Brassica napus	AGCAAGAAGATAGGTAGAGGGAAGATAGAGA
Glu3Term	TCTCTATCTTCCCTCTACCTATCTTCTTGCTACTCTCTGCATCGTGA	654
GAA-TAA	CTACTCCCACCTT A ATCCATTGCTTCTGTGATATCGTCACCAAATG
	AATAAACCACCGAAAAGAAGTGGGAGAG
	CAATGGAT T AAGGTGGG	655
	CCCACCTT A ATCCATTG	656

Male-sterile	TATTCATTTGGTGACGATATCACAGAAGCAATGGATGAAGGTGGG	657
AG	AGTAGTCACGATGCA T AGAGTAGCAAGAAGATAGGTAGAGGGAA
Brassica napus	GATAGAGATAAAGAGGATAGAGAACACAACAA
Glu11Term	TTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCTCTACCTATC	658
GAG-TAG	TTCTTGCTACTCT A TGCATCGTGACTACTCCCACCTTCATCCATTG
	CTTCTGTGATATCGTCACCAAATGAATA
	ACGATGCA T AGAGTAGC	659
	GCTACTCT A TGCATCGT	660

Male-sterile	GGTGACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCA	661
AG	CGATGCAGAGAGTAGC T AGAAGATAGGTAGAGGGAAGATAGAGA
Brassica napus	TAAAGAGGATAGAGAACACAACAAATCGTCAAG
Lys14Term	CTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTCCCT	662
AAG-TAG	GTACCTATCTTCT A GCTACTCTCTGCATCGTGACTACTCCCACCTT
	CATCCATTGCTTCTGTGATATCGTCACC
	AGAGTAGC T AGAAGATA	663
	TATCTTCT A GCTAGTCT	664

Male-sterile	GACGATATCACAGAAGCAATGGATGAAGGTGGGAGTAGTCACGA	665
AG	TGCAGAGAGTAGCAAG T AGATAGGTAGAGGGAAGATAGAGATAAA
Brassica napus	GAGGATAGAGAACACAACAAATCGTCAAGTAA
Lys15Term	TTACTTGACGATTTGTTGTGTTCTCTATCCTCTTTATCTCTATCTTC	666
AAG-TAG	CCTCTACCTATCT A CTTGCTACTCTCTGCATCGTGACTACTCCCAC
	CTTCATCCATTGCTTCTGTGATATCGTC
	GTAGCAAG T AGATAGGT	667
	ACCTATCT A CTTGCTAC	668

Male-sterile	CAACCAAAAAACTTAAAAATCTTCTCTTTCCTTTCCTTACAAGGTGA	669
AG	AGTAATGGACTTC T AAAGTGATCTAACCAGAGAGATCTCACCACAA
Lycopersicon	AGGAAACTAGGAAGGGGGAAAATTGAGA
esculentum	TCTCAATTTTCCCCCTTCCTAGTTTCCTTTGTGGTGAGATCTCTCT	670
Glu4Term	GGTTAGATCACTTT A GAAGTCCATTACTTCACCTTGTAAGGAAAGG
CAA-TAA	AAAGAGAAGATTTTTAAGTTTTTTGGTTG
	TGGACTTC+E,unc TAAAGTGAT	671
	ATCACTTT A GAAGTCCA	672

Male-sterile	AAAATCTTCTCTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCC	673
AG	AAAGTGATCTAACC T GAGAGATCTCACCACAAAGGAAACTAGGAA
Lycopersicon	GGGGGAAAATTGAGATCAAAAGGATCGAAA
esculentum	TTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTTTGT	674
Arg9Term	GGTGAGATCTCTC A GGTTAGATCACTTTGGAAGTCCATTACTTCAC
AGA-TGA	CTTGTAAGGAAAGGAAAGAGAAGATTTT
	ATCTAACC T GAGAGATC	675
	GATCTCTC A GGTTAGAT	676

Male-sterile	ATCTTCTCTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCCAAA	677
AG	GTGATCTAACCAGA T AGATCTCACCACAAAGGAAACTAGGAAGGG
Lycopersicon	GGAAAATTGAGATCAAAAGGATCGAAAACA
esculentum	TGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAGTTTCCTT	678
Glu10Term	TGTGGTGAGATCT A TCTGGTTAGATCACTTTGGAAGTCCATTACTT
GAG-TAG	CACCTTGTAAGGAAAGGAAAGAGAAGAT
	TAACCAGA T AGATCTCA	679
	TGAGATCT A TCTGGTTA	680

Male-sterile	CTTTCCTTTCCTTACAAGGTGAAGTAATGGACTTCCAAAGTGATCT	681
AG	AACCAGAGAGATCT G ACCACAAAGGAAACTAGGAAGGGGGAAAA
Lycopersicon	TTGAGATCAAAAGGATCGAAAACACGACGAA
esculentum	TTCGTCGTGTTTTCGATCCTTTTGATCTCAATTTTCCCCCTTCCTAG	682
Ser12Term	TTTCCTTTGTGGT C AGATCTCTGTGGTTAGATCACTTTGGAAGTCC
TCA-TGA	ATTACTTCACCTTGTAAGGAAAGGAAAG
	AGAGATCT G ACCACAAA	683
	TTTGTGGT C AGATCTCT	684

Male-sterile	GTACTCTCTATTTTCATCTTCCAACCCTTTCTTTCCTTACCAGGTGA	685
NAG1	AAGTATGGACTTC T AAAGTGATCTAACAAGAGAGATCTCTCCACAA
Nicotiana tabacum	AGGAAACTGGGAAGAGGAAAGATTGAGA
Gln4Term	TCTCAATCTTTCCTCTTCCCAGTTTCCTTTGTGGAGAGATCTCTCTT	686
CAA-TAA	GTTAGATCACTTT A GAAGTCCATACTTTCACCTGGTAAGGAAAGAA
	AGGGTTGGAAGATGAAAATAGAGAGTAC
	TGGACTTC T AAAGTGAT	687
	ATCACTTT A GAAGTCCA	688

Male-sterile	ATCTTCCAACCCTTTCTTTCCTTACCAGGTGAAAGTATGGACTTCC	689
NAG1	AAAGTGATCTAACA T GAGAGATCTCTCCACAAAGGAAACTGGGAA
Nicotiana tabacum	GAGGAAAGATTGAGATCAAACGGATCGAAA
Arg9Term	TTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTTTGT	690
AGA-TGA	GGAGAGATCTCTC A TGTTAGATCACTTTGGAAGTCCATACTTTCAC
	CTGGTAAGGAAAGAAAGGGTTGGAAGAT
	ATCTAACA T GAGAGATC	691
	GATCTCTC A TGTTAGAT	692

Male-sterile	TTCCAACCCTTTCTTTCCTTAGCAGGTGAAAGTATGGACTTCCAAA	693
NAG1	GTGATCTAACAAGA T AGATCTCTCCACAAAGGAAACTGGGAAGAG
Nicotiana tabacum	GAAAGATTGAGATCAAACGGATCGAAAACA
Glu10Term	TGTTTTCGATCCGTTTGATCTCAATCTTTCCTCTTCCCAGTTTCCTT	694
GAG-TAG	TGTGGAGAGATCT A TCTTGTTAGATGACTTTGGAAGTCCATACTTT
	CACCTGGTAAGGAAAGAAAGGGTTGGAA
	TAACAAGA T AGATCTCT	695
	AGAGATCT A TCTTGTTA	696

Male-sterile	CTTTCCTTACCAGGTGAAAGTATGGACTTCCAAAGTGATCTAACAA	697
NAG1	GAGAGATCTCTCCA T AAAGGAAACTGGGAAGAGGAAAGATTGAGA
Nicotiana tabacum	TCAAACGGATCGAAAACACAACGAATCGTC
Gln14Term	GACGATTCGTTGTGTTTTCGATCCGTTTGATCTCAATCTTTCCTCTT	698
CAA-TAA	CCCAGTTTCCTTT A TGGAGAGATCTCTCTTGTTAGATCACTTTGGA
	AGTCCATACTTTCACCTGGTAAGGAAAG
	TCTCTCCA T AAAGGAAA	699
	TTTCCTTT A TGGAGAGA	700

Male-sterile	GCCTATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAA	701
AG	AGAAGATTGGGAAGG T GAAAGATCGAGATCAAGCGGATCGAAAA
Rosa hybrida	CACCACCAATCGTCAAGTCACCTTCTGCAAAA
Gly22Term	TTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTCGATCCGCT	702
GGA-TGA	TGATCTGGATCTTTC A CCTTCCCAATCTTCTTTGGGCATCAGCGTC
	CAGGACCGTGTTGGGTTTGTTTTCATAGGC
	TGGGAAGG T GAAAGATC	703
	GATCTTTC A CCTTCCCA	704

Male-sterile	TATGAAAACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGA	705
AG	AGATTGGGAAGGGGA T AGATCGAGATCAAGCGGATCGAAAACAC
Rosa hybrida	CACCAATCGTCAAGTCACCTTCTGCAAAAGGC
Lys23Term	GCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTTCGATCC	706
AAG-TAG	GCTTGATCTCGATCT A TCCCCTTCCCAATCTTCTTTGGGCATCAGC
	GTCCAGGACCGTGTTGGGTTTGTTTTCATA
	GAAGGGGA T AGATCGAG	707
	CTCGATCT A TCCCCTTC	708

Male-sterile	AACAAACCCAACACGGTCCTGGACGCTGATGCCCAAAGAAGATTG	709
AG	GGAAGGGGAAAGATC T AGATCAAGCGGATCGAAAACACCACCAA
Rosa hybrida	TCGTCAAGTCACCTTCTGCAAAAGGCGCAATG
Glu25Term	CATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGGTGTTTT	710
GAG-TAG	CGATCCGCTTGATCT A GATCTTTCCCCTTCCCAATCTTCTTTGGGC
	ATCAGCGTCCAGGACCGTGTTGGGTTTGTT
	GAAAGATC T AGATCAAG	711
	CTTGATCT A GATCTTTC	712

Male-sterile	CCCAACACGGTCCTGGACGCTGATGCCCAAAGAAGATTGGGAAG	713
AG	GGGAAAGATCGAGATC T AGCGGATCGAAAACACCACCAATCGTCA
Rosa hybrida	AGTCACCTTCTGCAAAAGGCGCAATGGTTTGC
Lys27	GCAAACCATTGCGCCTTTTGCAGAAGGTGACTTGACGATTGGTGG	714
AAG-TAG	TGTTTTCGATCCGCT A GATCTCGATCTTTCCCCTTCCCAATCTTCT
	TTGGGCATCAGCGTCCAGGACCGTGTTGGG
	TCGAGATC T AGCGGATC	715
	GATCCGCT A GATCTCGA	716

Male-sterile	CAATTGCGTGTTTTTATTTTTTTTGTTTTTGACTAAGTAGAAATGGC	717
far	GTCTCTAAGCGAT T AATCGACCGAGGTATCGCGCGAGAGGAAAAT
Antirrhinum majus	CGGGAGAGGAAAGATCGAGATCAAACGGA
Gln7Term	TCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTCTCGGGCGA	718
CAA-TAA	TACCTCGGTCGATT A ATCGCTTAGAGACGCCATTTCTACTTAGTCA
	AAAAGAAAAAAAATAAAAACAGGCAATTG
	TAAGCGAT T AATCGACC	719
	GGTCGATT A ATCGCTTA	720

Male-sterile	GTTTTTATTTTTTTTCTTTTTGACTAAGTAGAAATGGCGTCTCTAAG	721
far	CGATCAATCGACC T AGGTATCGCCCGAGAGGAAAATCGGGAGAG
Antirrhinum majus	GAAAGATCGAGATCAAACGGATCGAAAACA
Glu10Term	TGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTCCCGATTTTCCTC	722
GAG-TAG	TCGGGCGATACCT A GGTCGATTGATCGCTTAGAGACGCCATTTCT
	ACTTAGTCAAAAAGAAAAAAAATAAAAAC
	AATCGACC T AGGTATCG	723
	CGATACCT A GGTCGATT	724

Male-sterile	TTTCTTTTTGACTAAGTAGAAATGGCGTCTCTAAGCGATCAATCGA	725
far	CCGAGGTATCGCCC T AGAGGAAAATCGGGAGAGGAAAGATCGAG
Antirrhinum majus	ATCAAACGGATCGAAAACAAAACAAATCAAC
Glu14Term	GTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTTCCTCTC	726
GAG-TAG	CCGATTTTCCTCT A GGGCGATACCTCGGTCGATTGATCGCTTAGA
	GACGCCATTTCTACTTAGTCAAAAAGAAA
	TATCGCCC T AGAGGAAA	727
	TTTCCTCT A GGGCGATA	728

Male-sterile	TTTGACTAAGTAGAAATGGCGTCTCTAAGCGATCAATCGACCGAG	729
far	GTATCGCCCGAGAGG T AAATCGGGAGAGGAAAGATCGAGATCAA
Antirrhinum majus	ACGGATCGAAAACAAAACAAATCAACAGGTTA
Lys16Term	TAACCTGTTGATTTGTTTTGTTTTCGATCCGTTTGATCTCGATCTTT	730
AAA-TAA	CCTCTCCCGATTT A CCTCTCGGGCGATACCTCGGTCGATTGATCG
	CTTAGAGACGCCATTTCTACTTAGTCAAA
	CCGAGAGG T AAATCGGG	731
	CCCGATTT A CCTCTCGG	732

Male-sterile	TGTCCAAGCATTATCAGTCACCACTCACAAGAATGATTAAGGAAGA	733
AG	AGGAAAGGGTAAGT A GCAAATAAAGGGGATGTTCCAGAATCAAGA
Cucumis sativus	AGAGAAGATGTCAGACTCGCCTCAGAGGAA
Leu21Term	TTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAACA	734
TTG-TAG	TCCCCTTTATTTGC T ACTTACCCTTTCCTTCTTCCTTAATCATTCTT
	GTGAGTGGTGACTGATAATGCTTGGACA
	GGGTAAGT A GCAAATAA	735
	TTATTTGC T ACTTACCC	736

Male-sterile	TCCAAGCATTATCAGTCACCACTCACAAGAATGATTAAGGAAGAA	737
AG	GGAAAGGGTAAGTTG T AAATAAAGGGGATGTTCCAGAATCAAGAA
Cucumis sativus	GAGAAGATGTCAGACTCGCCTCAGAGGAAGA
Gln22Term	TCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATTCTGGAA	738
CAA-TAA	CATCCCCTTTATTT A CAACTTACCCTTTCCTTCTTCCTTAATCATTC
	TTGTGAGTGGTGACTGATAATGCTTGGA
	GTAAGTTG T AAATAAAG	739
	CTTTATTT A CAACTTAC	740

Male-sterile	CATTATCAGTCACCACTCACAAGAATGATTAAGGAAGAAGGAAAG	741
AG	GGTAAGTTGCAAAT A TAGGGGATGTTCCAGAATCAAGAAGAGAAG
Cucumis sativus	ATGTCAGACTCGCCTCAGAGGAAGATGGGAA
Lys24Term	TTCCCATCTTCCTCTGAGGCGAGTCTGACATCTTCTCTTCTTGATT	742
AAG-TAG	CTGGAACATCCCCT A TATTTGCAACTTACCCTTTCCTTCTTCCTTAA
	TCATTCTTGTGAGTGGTGACTGATAATG
	TGCAAATA T AGGGGATG	743
	CATCCCCT A TATTTGCA	744

Male-sterile	CCACTCACAAGAATGATTAAGGAAGAAGGAAAGGGTAAGTTGCAA	745
AG	ATAAAGGGGATGTTC T AGAATCAAGAAGAGAAGATGTCAGACTCG
Cucumis sativus	CCTCAGAGGAAGATGGGAAGAGGAAAGATTG
Gln28Term	CAATCTTTCCTCTTCCCATCTTCCTCTGAGGCGAGTCTGACATCTT	746
CAG-TAG	CTCTTCTTGATTCT A GAACATCCCCTTTATTTGCAACTTACCCTTTC
	CTTCTTCCTTAATCATTCTTGTGAGTGG
	GGATGTTC T AGAATCAA	747
	TTGATTCT A GAACATCC	748

Male-sterile	CCACCACCACCACCACCACCACCACCACACCATGCTCAACATGAT	749
AG	GACTGATCTGAGCTG A GGGCCGTCGTCCAAGGTCAAGGAGCAGG
Zea mays	TGGCGGCGGCGCCGACGGGCTCCGGCGACAGG
Cys10Term	CCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCTGCTCCTTGA	750
TGC-TGA	CCTTGGACGACGGCCC T CAGCTCAGATCAGTCATCATGTTGAGCA
	TGGTGTGGTGGTGGTGGTGGTGGTGGTGGTGG
	CTGAGCTG A GGGCCGTC	751
	GACGGCCC T CAGCTCAG	752

Male-sterile	ACCACCACCACCACCACCACACCATGCTCAACATGATGACTGATC	753
AG	TGAGCTGCGGGCCGT A GTCCAAGGTCAAGGAGCAGGTGGCGGC
Zea mays	GGCGCCGACGGGCTCCGGCGACAGGCAGGGGCA
Ser13Term	TGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGCCACCT	754
TCG-TAG	GCTCCTTGACCTTGGAC T ACGGCCCGCAGCTCAGATCAGTCATCA
	TGTTGAGCATGGTGTGGTGGTGGTGGTGGTGGT
	CGGGCCGT A GTCCAAGG	755
	CCTTGGAC T ACGGCCCG	756

Male-sterile	CACCACCACCACCACACCATGCTCAACATGATGACTGATCTGAGC	757
AG	TGCGGGCCGTCGTCC T AGGTCAAGGAGCAGGTGGCGGCGGCGC
Zea mays	CGACGGGCTCCGGCGACAGGCAGGGGCAGGGGA
Lys15Term	TCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCCGCCGC	758
AAG-TAG	CACCTGCTCCTTGACCT A GGACGACGGCCCGCAGCTCAGATCAG
	TCATCATGTTGAGCATGGTGTGGTGGTGGTGGTG
	CGTCGTCC T AGGTCAAG	759
	CTTGACCT A GGACGACG	760

Male-sterile	CACCACCACACCATGCTCAACATGATGACTGATCTGAGCTGCGGG	761
AG	CCGTCGTCCAAGGTC T AGGAGCAGGTGGCGGCGGCGCCGACGG
Zea mays	GCTCCGGCGACAGGCAGGGGCAGGGGAGAGGCA
Lys17Term	TGCCTCTCCCCTGCCCCTGCCTGTCGCCGGAGCCCGTCGGCGCC	762
AAG-TAG	GCCGCCACCTGCTCCT A GACCTTGGACGACGGCCCGCAGCTCAG
	ATCAGTCATCATGTTGAGCATGGTGTGGTGGTG
	CCAAGGTC T AGGAGCAG	763
	CTGCTCCT A GACCTTGG	764

Male-sterile	TCCTACCTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACA	765
AG	AGAGCATGCACATC T GAGAAGAGGAGGCTACACCATCCACAGTAA
Zea mays	CAGGCATCATGTCGACCCTGACTTCGGCGG
Arg4Term	CCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTGGATGGT	766
CGA-TGA	GTAGCCTCCTCTTCTC A GATGTGCATGCTCTTGTTCCTATCACACA
	GATTTTGAGGTCTGAAGGAGAAAAGGTAGGA
	TGCACATC T GAGAAGAG	767
	CTCTTCTC A GATGTGCA	768

Male-sterile	TACCTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGA	769
AG	GCATGCACATCCGA T AAGAGGAGGCTACACCATCCACAGTAACAG
Zea mays	GCATCATGTCGACCCTGACTTCGGCGGGGC
Glu5Term	GCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACTGTGGAT	770
GAA-TAA	GGTGTAGCCTCCTCTT A TCGGATGTGCATGCTCTTGTTCCTATCAC
	ACAGATTTTGAGGTCTGAAGGAGAAAAGGTA
	ACATCCGA T AAGAGGAG	771
	CTCCTCTT A TCGGATGT	772

Male-sterile	CTTTTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCA	773
AG	TGCACATCCGAGAA T AGGAGGCTACACCATCCACAGTAACAGGCA
Zea mays	TCATGTCGACCCTGACTTCGGCGGGGCAGC
Glu6Term	GCTGCCCCGCCGAAGTGAGGGTCGACATGATGCCTGTTACTGTG	774
GAG-TAG	GATGGTGTAGCCTCCT A TTCTCGGATGTGCATGCTCTTGTTCCTAT
	CACACAGATTTTGAGGTCTGAAGGAGAAAAG
	TCCGAGAA T AGGAGGCT	775
	AGCCTCCT A TTCTCGGA	776

Male-sterile	TTCTCCTTCAGACCTCAAAATCTGTGTGATAGGAACAAGAGCATG	777
AG	CACATCCGAGAAGAG T AGGCTACACCATCCACAGTAACAGGCATC
Zea mays	ATGTCGACCCTGACTTCGGCGGGGCAGCAGA
Glu7Term	TCTGCTGCCCCGCCGAAGTCAGGGTCGACATGATGCCTGTTACT	778
GAG-TAG	GTGGATGGTGTAGCCT A CTCTTCTCGGATGTGCATGCTCTTGTTC
	CTATCACACAGATTTTGAGGTCTGAAGGAGAA
	GAGAAGAG T AGGCTACA	779
	TGTAGCCT A CTCTTCTC	780

Male-sterile	GCTGGGTCAGGATCGTCGGCGGCGGTGGCGGCGGGGAGCAGC	781
AG	GAGAAGATGGGGAGGGGG T AGATCGAGATAAAGCGGATCGAGAA
Oryza sativa	CACGACGAACCGGCAGGTGACCTTCTGCAAGCGCC
Lys5Term	GGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCTCGATC	782
AAG-TAG	CGCTTTATCTCGATCT A CCCCCTCCCCATCTTCTCGCTGCTCCCC
	GCCGCCACCGCCGCCGACGATCCTGACCCAGC
	GGAGGGGG T AGATCGAG	783
	CTCGATCT A CCCCCTCC	784

Male-sterile	TCAGGATCGTCGGCGGGGGTGGCGGCGGGGAGCAGCGAGAAGA	785
AG	TGGGGAGGGGGAAGATC T AGATAAAGCGGATCGAGAACACGACG
Oryza sativa	AACCGGCAGGTGACCTTCTGCAAGCGCCGCAATG
GTu7Term	CATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTCGTGTTCT	786
GAG-TAG	CGATCCGCTTTATCT A GATCTTCCCCCTCCCCATCTTCTCGCTGCT
	CCCCGCCGCCACCGCCGCCGACGATCCTGA
	GGAAGATC T AGATAAAG	787
	CTTTATCT A GATCTTCC	788

Male-sterile	TCGTCGGCGGCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGG	789
AG	AGGGGGAAGATCGAGATA T AGCGGATCGAGAACACGACGAACCG
Oryza sativa	GCAGGTGACCTTCTGCAAGCGCCGCAATGGCCTCC
Lys9Term	GGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGCCGGTTCGTC	790
AAG-TAG	GTGTTCTCGATCCGCT A TATCTCGATCTTCCCCCTCCCCATCTTCT
	CGCTGCTCCCCGCCGCCACCGCCGCCGACGA
	TCGAGATA T AGCGGATC	791
	GATCCGCT A TATCTCGA	792

Male-sterile	GCGGTGGCGGCGGGGAGCAGCGAGAAGATGGGGAGGGGGAAG	793
AG	ATCGAGATAAAGCGGATC T AGAACACGACGAACCGGCAGGTGAC
Oryza sativa	CTTCTGCAAGCGCCGCAATGGCCTCCTGAAGAAGG
Glu12Term	CCTTCTTCAGGAGGCCATTGCGGCGCTTGCAGAAGGTCACCTGC	794
GAG-TAG	CGGTTCGTCGTGTTCT A GATCCGCTTTATCTCGATCTTCCCCCTCC
	CCATCTTCTCGCTGCTCCCCGCCGCCACCGC
	AGCGGATC T AGAACACG	795
	CGTGTTCT A GATCCGCT	796

TABLE 16


Oligonucleotides to produce male-sterile plants

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Male-sterile	GGGAAGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAA	797
P1	TAGACAAGTTACATA G TCAAAGAGAAGAAATGGTATCATCAAAAAA
Cucumis sativus	GCCAAAGAAATTACTGTTCTTTGCGATGCT
Tyr21Term	AGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATACCAT	798
TAT-TAG	TTCTTCTCTTTGA C TATGTAACTTGTCTATTGCTTGAGTTCTCTATTC
	TTTTTATTTCTATTTTCCCTCTTCCC
	GTTACATA G TCAAAGAG	799
	CTCTTTGA C TATGTAAC	800

Male-sterile	GAAGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATA	801
P1	GACAAGTTACATATT G AAAGAGAAGAAATGGTATCATCAAAAAAGC
Cucumis sativus	CAAAGAAATTACTGTTCTTTGCGATGCTCA
Ser22Term	TGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATAC	802
TCA-TGA	CATTTCTTCTCTTT C AATATGTAACTTGTCTATTGCTTGAGTTCTCTA
	TTGTTTTTATTTCTATTTTCCCTCTTC
	TACATATT G AAAGAGAA	803
	TTCTCTTT+E,un CAATATGTA	804

Male-sterile	AGAGGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGAC	805
P1	AAGTTAGATATTCA T AGAGAAGAAATGGTATCATCAAAAAAGCCAA
Cucumis sativus	AGAAATTACTGTTCTTTGCGATGCTCAAG
Lys23Term	CTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATGATA	806
AAG-TAG	CCATTTCTTCTCT A TGAATATGTAACTTGTCTATTGCTTGAGTTCTC
	TATTCTTTTTATTTCTATTTTCCCTCT
	CATATTCA T AGAGAAGA	807
	TCTTCTCT A TGAATATG	808

Male-sterile	GGGAAAATAGAAATAAAAAGAATAGAGAACTCAAGCAATAGACAAG	809
P1	TTACATATTCAAAG T GAAGAAATGGTATCATCAAAAAAGCCAAAGA
Cucumis sativus	AATTACTGTTCTTTGCGATGCTCAAGTTT
Arg24Term	AAACTTGAGCATCGCAAAGAACAGTAATTTCTTTGGCTTTTTTGATG	810
AGA-TGA	ATACCATTTCTTC A CTTTGAATATGTAACTTGTCTATTGCTTGAGTT
	CTCTATTCTTTTTATTTCTATTTTCCC
	ATTCAAAG T GAAGAAAT	811
	ATTTCTTC A CTTTGAAT	812

Male-sterile	GGGACGTGGGAAGGTTGAGATCAAGAGGATTGAGAACTGAAGTAA	813
P1	CAGGCAGGTGACCTA G TCCAAGAGGAGGAATGGGATTATCAAGAA
Malus domestica	GGCAAAGGAGATCACTGTTCTATGTGATGCT
Tyr21Term	AGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATCCCA	814
TAG-TAG	TTCCTCCTCTTGGA C TAGGTGACCTGCCTGTTACTTGAGTTCTCAA
	TCCTCTTGATCTCAACCTTCCCACGTCGC
	GTGACCTA G TGCAAGAG	815
	CTCTTGGA C TAGGTCAC	816

Male-sterile	CGTGGGAAGGTTGAGATCAAGAGGATTGAGAACTCAAGTAACAGG	817
P1	CAGGTGACCTACTCC T AGAGGAGGAATGGGATTATCAAGAAGGCA
Malus domestica	AAGGAGATCACTGTTCTATGTGATGCTAAAG
Lys23Term	CTTTAGCATCACATAGAACAGTGATCTCCTTTGCCTTCTTGATAATC	818
AAG-TAG	CCATTCCTCCTCT A GGAGTAGGTCACCTGCCTGTTACTTGAGTTCT
	CAATCCTCTTGATCTCAACCTTCCCACG
	CCTACTCC T AGAGGAGG	819
	CCTCCTCT A GGAGTAGG	820

Male-sterile	AGGATTGAGAAGTCAAGTAACAGGCAGGTGACCTACTCCAAGAGG	821
P1	AGGAATGGGATTATC T AGAAGGCAAAGGAGATGACTGTTCTATGT
Malus domestica	GATGCTAAAGTATCTCTTATCATTTATTCTA
Lys30Term	TAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGTGAT	822
AAG-TAG	CTCCTTTGCCTTCT A GATAATCGCATTCCTCCTCTTGGAGTAGGTC
	ACCTGCCTGTTACTTGAGTTCTCAATCCT
	GGATTATC T AGAAGGCA	823
	TGCCTTCT A GATAATCC	824

Male-sterile	ATTGAGAACTCAAGTAACAGGCAGGTGACCTACTCCAAGAGGAGG	825
P1	AATGGGATTATCAAG T AGGCAAAGGAGATCACTGTTCTATGTGATG
Malus domestica	CTAAAGTATCTCTTATCATTTATTCTAGCT
Lys31Term	AGCTAGAATAAATGATAAGAGATACTTTAGCATCACATAGAACAGT	826
AAG-TAG	GATCTCCTTTGCCT A CTTGATAATGCCATTCCTCCTCTTGGAGTAG
	GTCACCTGCCTGTTACTTGAGTTCTCAAT
	TTATCAAG T AGGCAAAG	827
	CTTTGCCT A CTTGATAA	828

Male-sterile	CATTTTTACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAA	829
globosa	AAACAAAAAAATG T GAAGAGGAAAAATTGAGATCAAAAGAATTGAG
Antirrhinum majus	AACTCAAGCAACAGGCAGGTTACTTACT
Gly2Term	AGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTGATCTCA	830
GGA-TGA	ATTTTTCCTCTTC A CATTTTTTTGTTTTTGTTTTTCTCTCTTGTTTTTG
	TTTGCAGATAACTATTGTAAAAATG
	AAAAAATG T GAAGAGGA	831
	TCCTCTTC A CATTTTTT	832

Male-sterile	TTTTACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAA	833
globosa	CAAAAAAATGGGA T GAGGAAAAATTGAGATCAAAAGAATTGAGAAC
Antirrhinum majus	TCAAGCAACAGGCAGGTTACTTACTCAA
Arg3Term	TTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTGATC	834
AGA-TGA	TCAATTTTTCCTC A TCCCATTTTTTTGTTTTTGTTTTTCTCTCTTGTTT
	TTGTTTGCAGATAACTATTGTAAAA
	AAATGGGA T GAGGAAAA	835
	TTTTCCTC A TCCCATTT	836

Male-sterile	TACAATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACA	837
globosa	AAAAAATGGGAAGA T GAAAAATTGAGATCAAAAGAATTGAGAACTC
Antirthinum majus	AAGCAACAGGCAGGTTACTTACTCAAAGA
Gly4Term	TCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTCTTTTG	838
GGA-TGA	ATCTCAATTTTTC A TCTTCCCATTTTTTTGTTTTTGTTTTTCTCTCTTG
	TTTTTGTTTGCAGATAACTATTGTA
	TGGGAAGA T GAAAAATT	839
	AATTTTTC A TCTTCCCA	840

Male-sterile	AATAGTTATCTGCAAACAAAAACAAGAGAGAAAAACAAAAACAAAA	841
globosa	AAATGGGAAGAGGA T AAATTGAGATCAAAAGAATTGAGAACTCAAG
Antirrhinum majus	CAACAGGCAGGTTACTTACTCAAAGAGAA
Lys5Term	TTCTCTTTGAGTAAGTAACCTGCCTGTTGCTTGAGTTCTCAATTGTT	842
AAA-TAA	TTGATCTCAATTT A TCCTCTTCCCATTTTTTTGTTTTTGTTTTTCTCT
	CTTGTTTTTGTTTGCAGATAACTATT
	GAAGAGGA T AAATTGAG	843
	CTCAATTT A TCCTCTTC	844

Male-sterile	GCTGAGCTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGC	845
P1	AGTATGGGGCGCGGC T AGATCAAGATCAAGAGGATCGAGAACTCT
Zea mays	ACCAACCGGCAGGTGACCTTCTCCAAGCGCC
Lys5Term	GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC	846
AAG-TAG	TCTTGATCTTGATCT A GCCGCGCCCCATACTGCGTTCTCCACTCCC
	AAACAGATCCAAGGGCAGCAAGAGCTCAGC
	GGCGCGGC T AGATGAAG	847
	CTTGATCT A GCCGCGCC	848

Male-sterile	CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG	849
P1	GGGCGCGGCAAGATC T AGATCAAGAGGATCGAGAACTCTACCAAC
Zea mays	CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG
Lys7Term	CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC	850
AAG-TAG	TCGATCCTCTTGATCT A GATCTTGCCGCGCCCCATACTGCGTTCTC
	CACTCCCAAACAGATCCAAGGGCAGCAAGAG
	GCAAGATC T AGATCAAG	851
	CTTGATCT A GATCTTGC	852

Male-sterile	CTCTTGCTGCCCTTGGATCTGTTTGGGAGTGGAGAACGCAGTATG	853
P1	GGGCGCGGCAAGATC T AGATCAAGAGGATCGAGAACTCTACCAAC
Zea mays	CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG
Lys9Term	CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC	854
AAG-TAG	TCGATCCTCTTGATCT A GATCTTGCCGCGCCCCATACTGCGTTCTC
	CACTCCCAAACAGATCCAAGGGCAGCAAGAG
	GCAAGATC T AGATCAAG	855
	GTTGATCT A GATCTTGC	856

Male-sterile	GATCTGTTTGGGAGTGGAGAACGCAGTATGGGGCGCGGCAAGAT	857
P1	CAAGATCAAGAGGATC T AGAACTCTACCAACCGGCAGGTGACCTT
Zea mays	CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG
Glu12Term	CCTTCTTGACGAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC	858
GAG-TAG	CGGTTGGTAGAGTTCT A GATCCTCTTGATCTTGATCTTGCCGCGCC
	CCATACTGCGTTCTCCACTCCCAAACAGATC
	AGAGGATC T AGAACTCT	859
	AGAGTTCT A GATGCTCT	860

Male-sterile	GCTGAGCTCTTGCTGCCCTTGAATCTGTTAGGGAGTGGAGAACGG	861
P1	AGTATGGGGCGCGGC T AGATCGAGATCAAGAGGATCGAGAACTCT
Zea mays	ACCAACCGGCAGGTGACCTTCTCCAAGCGCC
Lys5Term	GGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTCTCGATCC	862
AAG-TAG	TCTTGATCTCGATCT A GCCGCGCCCCATACTCCGTTCTCCACTCCC
	TAACAGATTCAAGGGCAGCAAGAGCTCAGC
	GGCGCGGC T AGATCGAG	863
	CTCGATCT A GCCGCGCC	864

Male-sterile	CTCTTGCTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATG	865
P1	GGGCGCGGCAAGATC T AGATCAAGAGGATCGAGAACTCTACCAAC
Zea mays	CGGCAGGTGACCTTCTCCAAGCGCCGGGCCG
Glu7Term	CGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTAGAGTTC	866
GAG-TAG	TCGATCCTCTTGATCT A GATCTTGCCGCGCCCCATACTCCGTTCTC
	CACTCCCTAACAGATTCAAGGGCAGCAAGAG
	GCAAGATC T AGATCAAG	867
	CTTGATCT A GATCTTGC	868

Male-sterile	CTGCCCTTGAATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCG	869
P1	CGGCAAGATCGAGATC T AGAGGATCGAGAACTCTACCAACCGGCA
Zea mays	GGTGACCTTCTCCAAGCGCCGGGCCGGACTGG
Lys9Term	CCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGCCGGTTGGTA	870
AAG-TAG	GAGTTCTCGATCCTCT A GATCTCGATCTTGCCGCGCCCCATACTC
	CGTTCTCCACTCCCTAACAGATTCAAGGGCAG
	TCGAGATC T AGAGGATC	871
	GATCCTCT A GATCTCGA	872

Male-sterile	AATCTGTTAGGGAGTGGAGAACGGAGTATGGGGCGCGGCAAGAT	873
P1	GGAGATGAAGAGGATC T AGAACTCTACCAACCGGCAGGTGACCTT
Zea mays	CTCCAAGCGCCGGGCCGGACTGGTCAAGAAGG
Glu12Term	CCTTCTTGACCAGTCCGGCCCGGCGCTTGGAGAAGGTCACCTGC	874
GAG-TAG	CGGTTGGTAGAGTTCT A GATCCTCTTGATCTCGATCTTGCCGCGC
	CCCATACTCCGTTCTCCACTCCCTAACAGATT
	AGAGGATC T AGAACTCT	875
	AGAGTTCT A GATCCTCT	876

Male-sterile	TTGCTGCTAAGCTAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGG	877
P1	CGGGATGGGGCGCGGG+E,un TAGATCGAGATCAAGAGGATCGAGAACT
Oryza sativa	CCACCAACCGCCAGGTGACCTTCTCCAAGCGCA
Lys5Term	TGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCTCGATCC	878
AAG-TAG	TCTTGATGTCGATGT A CCCGCGCCCCATCCCGCCTCCTCCTCCTC
	CTCCTCCTTCCTCCAGCTAGCTTAGCAGCAA
	GGCGCGGG T AGATCGAG	879
	CTCGATCT A CCCGCGCC	880

Male-sterile	CTAAGCTAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGA	881
P1	TGGGGCGCGGGAAGATC T AGATCAAGAGGATCGAGAACTCCACC
Oryza sativa	AACCGCCAGGTGACCTTCTCCAAGCGCAGGAGCG
Glu7Term	CGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTGGAGTTCT	882
GAG-TAG	CGATCCTCTTGATCT A GATCTTCCCGCGCCCCATCCCGCCTCCTC
	CTCCTCCTCCTCCTTCCTCCAGCTAGCTTAG
	GGAAGATC T AGATCAAG	883
	CTTGATCT A GATCTTCC	884

Male-sterile	TAGCTGGAGGAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGC	885
P1	GCGGGAAGATCGAGATC T AGAGGATCGAGAACTCCACCAACCGC
Oryza sativa	CAGGTGACCTTCTCCAAGCGCAGGAGCGGGATCC
Lys9Term	GGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGCGGTTGGTG	886
AAG-TAG	GAGTTCTCGATCCTCT A GATCTCGATCTTCCCGCGCCCCATCCCG
	CCTCCTCCTCCTCCTCCTCCTTCCTCCAGCTA
	TCGAGATC T AGAGGATC	887
	GATCCTCT A GATCTCGA	888

Male-sterile	GAAGGAGGAGGAGGAGGAGGAGGCGGGATGGGGCGCGGGAAG	889
P1	ATCGAGATCAAGAGGATC T AGAACTCCACCAACCGCCAGGTGACC
Oryza sativa	TTCTCCAAGCGCAGGAGCGGGATCCTCAAGAAGG
Glu12Term	CCTTCTTGAGGATCCCGCTCCTGCGCTTGGAGAAGGTCACCTGGC	890
GAG-TAG	GGTTGGTGGAGTTCT A GATCCTCTTGATCTCGATCTTCCCGCGCC
	CCATCCCGCCTCCTCCTCCTCCTCCTCCTTC
	AGAGGATC T AGAACTCC	891
	GGAGTTCT A GATCCTCT	892

EXAMPLE 7

Engineering Plants for Abiotic Stress Tolerance

Environmental stresses, such as drought, increased soil salinity, soil contamination with heavy meals, and extreme temperature, are major factors limiting plant growth and productivity. The worldwide loss in yield of three major cereal crops, rice, maize, and wheat due to water stress (drought) has been estimated to be over ten billion dollars annually and many currently marginal soils could be brought into cultivation if suitable plant varieties were available. [0128]
Physiological and biochemical responses to high levels of ionic or nonionic solutes and decreased water potential have been studied in a variety of plants. It is known, for example, that increasing levels of alcohol dehydrogenase can confer enhances flooding resistance in plants. There are also several possible mechanisms to enhance plant salt tolerance. For example, one mechanism underlying the adaptation or tolerance of plants to osmotic stresses is the accumulation of compatible, low molecular weight osmolytes such as sugar alcohols, special amino acids, and glycinebetaine. Such accumulation can be engineered, for example, by removing feedback inhibition on 1-pyrroline-t-carboxylate synthetase, which results in accumulation of proline. Additionally, recent experiments suggest that altering the expression or activity of specific sodium or potassium transporters can confer enhanced salt tolerance. [0129]
Plant tolerance of contamination by heavy metals such as lead and aluminum in soils has also been investigated and one mechanism underlying tolerance is the production of dicarboxylic acids such as oxalate and citrate. In addition, individual genes involved in heavy metal sensitivity have been identified. [0130]

The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that confer stress tolerance in plants.

TABLE 17


Genome-Altering Oligos Conferring Stress Tolerance

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Salt Tolerance	CGTCTTTTTGTGTGGTAGTTGGATGTGACGGTTGCTCAAATGCTT	893
P5CS	GTGACCGATAGCAGT GC TAGAGATAAGGATTTCAGGAAGCAACTT
Arabidopsis thaliana	AGTGAAACTGTCAAAGCGATGCTGAGGATGA
Phe128Ala	TCATCCTCAGCATCGCTTTGACAGTTTCACTAAGTTGCTTCCTGAA	894
TTT-GCT	ATCCTTATGTCTA GC ACTGCTATCGGTCACAAGCATTTGAGCAACC
	GTCACATCCAACTACCACACAAAAAGACG
	ATAGCAGT GC TAGAGAT	895
	ATCTCTA GC ACTGCTAT	896

Salt Tolerance	GAGAGTATGTTTGACCAGCTGGATGTGACGGCTGCTCAGCTGCTG	897
P5CS 1	GTGAATGACAGTAGT GC CAGAGACAAGGAGTTCAGGAAGCAACTT
Brassica napus	AATGAGACAGTGAAGTCCATGCTTGATTTGA
Phe128Ala	TCAAATCAAGCATGGACTTCACTGTCTCATTAAGTTGCTTCCTGAA	898
TTC-GCC	CTCCTTGTCTCTG GC ACTACTGTCATTCACCAGCAGCTGAGCAGC
	CGTCACATCCAGCTGGTCAAACATAGTGTC
	ACAGTAGT GC CAGAGAC	899
	GTCTCTG GC ACTACTGT	900

Salt Tolerance	GAGACTATGTTTGACCAGATGGATGTGACGGTGGCTCAAATGCTG	901
P505 2	GTGACTGATAGCAGT G TCAGAGATAAGGATTTCAGGAAGCAACTT
Brassica napus	AGTGAGACAGTCAAAGCTATGCTGAAAATGA
Phe129Ala	TCATTTTCAGCATAGCTTTGACTGTCTCACTAAGTTGCTTCCTGAA	902
TTC-GCC	ATCCTTATCTCTGA C ACTGCTATCAGTCACCAGCATTTGAGCCACC
	GTCACATCCATCTGGTCAAACATAGTCTC
	ATAGCAGT G TCAGAGAT	903
	ATCTCTGA C ACTGCTAT	904

Salt Tolerance	GATATGTTGTTTAACCAACTGGATGTCTCGTCATCTCAACTTCTTG	905
P5GS	TCACCGACAGTGAT GC TGAGAACCCAAAGTTCCGGGAGCAACTCA
Oryza sativa	CTGAAACTGTTGAGTCATTATTAGATCTTA
Phe128Ala	TAAGATCTAATAATGACTCAACAGTTTCAGTGAGTTGCTCCCGGAA	906
TTT-GCT	CTTTGGGTTCTCA GC ATCACTGTCGGTGACAAGAAGTTGAGATGA
	CGAGACATCCAGTTGGTTAAACAACATATC
	ACAGTGAT GC TGAGAAC	907
	GTTCTCA GC ATCACTGT	908

Salt Tolerance	GATATTTTGTTTAGTCAGCTGGATGTGACATCTGCTCAGCTTCTTG	909
P5CS	TTACTGACAATGAT GC TAGAGACCAAGATTTTAGAAAGCAACTTTC
Medicago sativa	TGAAACTGTGAGATCACTTCTAGCACTAA
Phe128Ala	TTAGTGCTAGAAGTGATCTCACAGTTTCAGAAAGTTGCTTTCTAAA	910
TTT-GCT	ATCTTGGTCTCTA GC ATCATTGTCAGTAAGAAGAAGCTGAGCAGAT
	GTCACATCCAGCTGACTAAACAAAATATC
	ACAATGAT GC TAGAGAC	911
	GTCTCTA GC ATCATTGT	912

Salt Tolerance	GATACATTGTTTAGTCAGCTGGATGTGACATCAGCTCAGCTACTC	913
P5CS	GTTACTGATAATGAT GC TAGGGATCCAGAATTCAGGAAGCAACTT
Actinidia deliciosa	ACTGAAACTGTAGAATCACTATTGAATTTGA
Phe128Ala	TCAAATTCAATAGTGATTCTACAGTTTCAGTAAGTTGCTTCCTGAAT	914
TTT-GCT	TCTGGATCCCTA GC ATCATTATCAGTAACGAGTAGCTGAGCTGAT
	GTCACATCCAGCTGACTAAACAATGTATC
	ATAATGAT GC TAGGGAT	915
	ATCCCTA GC ATCATTAT	916

Salt Tolerance	GACACACTCTTCAGTCAACTGGATGTGACATCAGCACAGCTTCTT	917
P5CS	GTAACAGATAATGAC GC CAGAAGTCCAGAATTTAGAAAACAACTTA
Cichorium intybus	CTGAAACAGTCGATTCTTTATTATCTTATA
Phe122Ala	TATAAGATAATAAAGAATCGACTGTTTCAGTAAGTTGTTTTCTAAAT	918
TTC-GCC	TCTGGACTTCTG GC GTCATTATCTGTTACAAGAAGCTGTGCTGAT
	GTCACATCCAGTTGACTGAAGAGTGTGTC
	ATAATGAC GC CAGAAGT	919
	ACTTCTG GC GTCATTAT	920

Salt Tolerance	GATTCTTTGTTCAGTCAGTTGGATGTGACATCAGCTCAGCTTCTGG	921
P5CS	TGACTGATAATGAC GC TAGAGATCCAGATTTTAGGAGACAACTCA
Lycopersicon	ATGACACAGTAAATTCGTTGCTTTCTCTAA
esculentum	TTAGAGAAAGCAACGAATTTACTGTGTCATTGAGTTGTCTCCTAAA	922
Phe12BAla	ATCTGGATCTCTA GC GTCATTATCAGTCACCAGAAGCTGAGCTGA
TTT-GCT	TGTCACATCCAACTGACTGAACAAAGAATC
	ATAATGAC GC TAGAGAT	923
	ATCTCTA GC GTCATTAT	924

Salt Tolerance	GATACCATGTTCAGCCAGCTTGATGTGACTTCTTCCCAACTTCTTG	925
P5CS	TGAATGATGGATTT GC TAGGGATGCTGGCTTCAGAAAACAACTTT
Vigna unguiculata	CGGACACAGTGAACGCGTTATTAGATTTAA
Phe162Ala	TTAAATCTAATAACGCGTTCACTGTGTCCGAAAGTTGTTTTCTGAA	926
TTT-GCT	GCCAGCATCCCTA GC AAATCCATCATTCACAAGAAGTTGGGAAGA
	AGTCACATCAAGCTGGCTGAACATGGTATC
	ATGGATTT GC TAGGGAT	927
	ATCCCTA GC AAATCCAT	928

Salt Tolerance	GACACCTTGTTTAGTCAGTTGGATCTGACTGCTGCTCAGCTGCTT	929
P5CS	GTGACGGACAACGAC GC TAGAGATCCAAGTTTTAGAACACAACTA
Mesembryanthemum	ACTGAAACAGTGTATCAGTTGTTGGATCTAA
crystallinum	TTAGATCCAACAACTGATACACTGTTTCAGTTAGTTGTGTTCTAAA	930
Phe125Ala	ACTTGGATCTCTA GC GTCGTTGTCCGTCACAAGCAGCTGAGCAGC
TTT-GCT	AGTCAGATCCAACTGACTAAACAAGGTGTC
	ACAACGAC GC TAGAGAT	931
	ATCTCTA GC GTCGTTGT	932

Salt Tolerance	GACACATTATTTAGCCAGCTGGATGTGACATCAGCTCAGCTTCTT	933
P5CS	GTGACTGATAATGAT GC TAGGGATGAAGCTTTCCGAAATCAACTTA
Vitis vinifera	CTCAAACAGTGGATTCATTGTTAGCTTTGA
Phe130Ala	TCAAAGCTAACAATGAATCCACTGTTTGAGTAAGTTGATTTCGGAA	934
TTT-GCT	AGCTTCATCCCTA GC ATCATTATCAGTCACAAGAAGCTGAGCTGAT
	GTCACATCCAGCTGGCTAAATAATGTGTC
	ATAATGAT GC TAGGGAT	935
	ATCCCTA GC ATCATTAT	936

Salt Tolerance	GATACGCTGTTCACTCAGCTCGATGTGACATCGGCTCAGCTTCTT	937
P5CS	GTGACGGATAACGAT GC TCGAGATAAGGATTTCAGGAAGCAGCTT
Vigna aconitifolia	ACTGAGACTGTGAAGTCGCTGTTGGGGCTGA
Phe129Ala	TCAGCGCCAACAGCGACTTCACAGTCTCAGTAAGCTGCTTCCTGA	938
TTT-GCT	AATCCTTATCTCGA GC ATCGTTATCCGTCACAAGAAGCTGAGCCG
	ATGTCACATCGAGCTGAGTGAACAGCGTATC
	ATAACGAT GC TCGAGAT	939
	ATCTCGA GC ATCGTTAT	940

Salt Tolerance	AGAGATGTTCTTAGTTCCAAAGAAATCTCACCTCTCAGTTTCTCCG	941
HKT1	TCTTCACAACAGTT GT CACGTTTGCAAACTGCGGATTTGTCCCCAC
Arabidopsis thaliana	GAATGAGAACATGATCATCTTTCGCAAAA
Ser207Val	TTTTGCGAAAGATGATCATGTTCTCATTCGTGGGGACAAATCCGC	942
TCC-GTC	AGTTTGCAAACGTG AC AACTGTTGTGAAGACGGAGAAAGTGAGAG
	GTGAGATTTCTTTGGAACTAAGAACATCTCT
	CAACAGTT GT CACGTTT	943
	AAACGTG AC AACTGTTG	944

Salt Tolerance	CGAATGAGAACATGATCATCTTTCGCAAAAACTCTGGTCTCATCTG	945
HKT1	GCTCCTAATCCCTC T AGTACTGATGGGAAACACTTTGTTCCCTTGC
Arabidopsis thaliana	TTCTTGGTTTTGCTCATATGGGGACTTTA
Gln237Leu	TAAAGTCCCCATATGAGCAAAACCAAGAAGCAAGGGAACAAAGTG	946
CAA-CTA	TTTCCCATCAGTACT A GAGGGATTAGGAGCCAGATGAGACCAGAG
	TTTTTGCGAAAGATGATCATGTTCTCATTCG
	AATCCCTC T AGTACTGA	947
	TCAGTACT A GAGGGATT	948

Salt Tolerance	AGTCTCTAGAAGGAATGAGTTCGTACGAGAAGTTGGTTGGATCGT	949
HKT1	TGTTTCAAGTGGTGA G TTCGCGACACACCGGAGAAACTATAGTAG
Arabidopsis thaliana	ACCTCTCTACACTTTCCCCAGCTATCTTGGT
Asn332Ser	ACCAAGATAGCTGGGGAAAGTGTAGAGAGGTCTACTATAGTTTCT	950
AAT-AGT	CCGGTGTGTCGCGAA C TCACCACTTGAAACAACGATCCAACCAAC
	TTCTCGTACGAACTCATTCCTTCTAGAGACT
	AGTGGTGA G TTCGCGAC	951
	GTCGCGAA C TCACCACT	952

Salt Tolerance	AGAGATGTGCTAAAGAAGAAAGGTCTCAAAATGGTGACCTTTTCC	953
HKT1	GTCTTCACCACCGTG GT GACCTTTGCCAGTTGTGGGTTTGTCCCG
Eucalyptus	ACCAATGAAAACATGATTATCTTCAGCAAAA
camaldulensis	TTTTGCTGAAGATAATCATGTTTTCATTGGTCGGGACAAACCCACA	954
Ser256Val	ACTGGCAAAGGTC AC CACGGTGGTGAAGACGGAAAAGGTCACCA
TCG-GTG	TTTTGAGACCTTTCTTCTTTAGCACATCTCT
	CCACCGTG GT GACCTTT	955
	AAAGGTC AC CACGGTGG	956

Salt Tolerance	CCAATGAAAACATGATTATCTTCAGCAAAAACTCTGGCCTCCTCCT	957
HKT1	GATTCTCATCCCTC T GGCCCTTCTTGGGAACATGCTGTTCCCATC
Eucalyptus	GAGCCTACGTTTGACGCTTTGGCTCATCGG
camaldulensis	CCGATGAGCCAAAGCGTCAAACGTAGGCTCGATGGGAACAGCAT	958
Gln286Leu	GTTCCCAAGAAGGGCCA G AGGGATGAGAATCAGGAGGAGGCCA
CAG-CTG	GAGTTTTTGCTGAAGATAATCATGTTTTCATTGG
	CATCCCTC T GGCCCTTC	959
	GAAGGGCC A GAGGGATG	960

Salt Tolerance	AATCGTTGAATGGACTAAGCTCCTGTGAGAAAATCGTGGGCGCGC	961
HKT1	TGTTTCAGTGCGTGA G CAGCAGACATACCGGCGAGACGGTCGTC
Eucalyptus	GATCTGTCCACAGTTGCTCCCGCCATCTTGGT
camaldulensis	ACCAAGATGGCGGGAGCAACTGTGGACAGATCGACGACCGTCTC	962
Asn381Ser	GCCGGTATGTCTGCTG C TCACGCACTGAAACAGCGCGCCCACGA
AAC-AGC	TTTTCTCACAGGAGCTTAGTCCATTCAACGATT
	GTGCGTGA G CAGCAGAC	963
	GTCTGCTG+E,un CTCACGCAC	964

Salt Tolerance	AAAGCTCCACTGAAGAAGAAAGGGATCAACATTGCACTCTTCTCA	965
HKT1	TTCTCGGTCACGGTC GT CTCGTTTGCGAATGTGGGGCTCGTGCC
Oryza sativa	GACAAATGAGAACATGGCAATCTTCTCCAAGA
Ser238Val	TCTTGGAGAAGATTGCCATGTTCTCATTTGTCGGCACGAGCCCCA	966
TCC-GTC	CATTCGCAAACGAG AC GACCGTGACCGAGAATGAGAAGAGTGCA
	ATGTTGATCCCTTTCTTCTTCAGTGGAGCTTT
	TCACGGTC GT CTCGTTT	967
	AAACGAG AC GACCGTGA	968

Salt Tolerance	CAAATGAGAACATGGCAATCTTCTCCAAGAACCCGGGCCTCCTCC	969
HKT1	TCCTGTTCATCGGCC T GATTGTTGCAGGCAATACACTTTACCCTCT
Oryza sativa	CTTCCTAAGGCTATTGATATGGTTCCTGGG
Gln268Leu	CCCAGGAACCATATCAATAGCCTTAGGAAGAGAGGGTAAAGTGTA	970
CAG-CTG	TTGCCTGCAAGAATC A GGCCGATGAACAGGAGGAGGAGGCCCGG
	GTTCTTGGAGAAGATTGCCATGTTCTCATTTG
	CATCGGCC T GATTCTTG	971
	CAAGAATC A GGCCGATG	972

Salt Tolerance	CAGTCTTTGATGGACTCAGCTCTTACCAGAAGATTATCAATGCATT	973
HKT1	GTTCATGGCAGTGA G CGCAAGGCACTCGGGGGAGAACTCCATCG
Oryza sativa	ACTGCTCACTCATCGCCCCTGCTGTTCTAGT
Asn363Ser	ACTAGAACAGCAGGGGCGATGAGTGAGCAGTCGATGGAGTTCTC	974
AAC-AGC	CCCCGAGTGCCTTGCG C TCACTGCCATGAACAATGCATTGATAAT
	CTTCTGGTAAGAGCTGAGTCCATCAAAGACTG
	GGCAGTGA G CGCAAGGC	975
	GCCTTGCG C TCACTGCC	976

Salt Tolerance	GTGCCCCACTGAACAAGAAAGGGATCAACATCGTGCTCTTCTCAC	977
HKT1	TATCAGTCACCGTTG T CTCCTGTGCGAATGCAGGACTCGTGCCCA
Triticum aestivum	CAAATGAGAACATGGTCATCTTCTCAAAGAA
Ala240Val	TTCTTTGAGAAGATGACCATGTTCTCATTTGTGGGCACGAGTCCT	978
GCC-GTC	GCATTCGCACAGGAG A CAACGGTGAGTGATAGTGAGAAGAGCAC
	GATGTTGATCCCTTTCTTGTTCAGTGGGGCAC
	CACCGTTG T CTCCTGTG	979
	CACAGGAG A CAACGGTG	980

Salt Tolerance	CAAATGAGAACATGGTCATCTTCTCAAAGAATTCAGGCCTCTTGTT	981
HKT1	GCTGCTGAGTGGCC T GATGCTCGCAGGCAATACATTGTTCCCTCT
Triticum aestivum	CTTCCTGAGGCTACTGGTGTGGTTCCTGGG
Gln270Leu	CCCAGGAACCACACCAGTAGCCTCAGGAAGAGAGGGAACAATGT	982
CAG-CTG	ATTGCCTGCGAGCATC A GGCCACTCAGCAGCAACAAGAGGCCTG
	AATTCTTTGAGAAGATGACCATGTTCTCATTTG
	GAGTGGCC T GATGCTCG	983
	CGAGCATC A GGCCACTC	984

Salt Tolerance	CAGTCTTTGATGGGCTCAGCTCTTATCAGAAGACTGTCAATGCATT	985
HKT1	CTTCATGGTGGTGA G TGCGAGGCACTCAGGGGAGAATTCCATCG
Triticum aestivum	ACTGCTCGCTCATGTCCCCTGCCATTATAGT
Asn365Ser	ACTATAATGGCAGGGGACATGAGCGAGCAGTCGATGGAATTCTCC	986
AAT-AGT	CCTGAGTGCCTCGCA C TCACCACCATGAAGAATGCATTGACAGTC
	TTCTGATAAGAGCTGAGCCCATCAAAGACTG
	GGTGGTGA G TGCGAGGC	987
	GCCTCGCA C TCACCACC	988

Freezing Tolerance	TTTTTTTTGTTTTCGTTTTCAAAAAGAAAATCTTTGAATTTTATGGCA	989
praline oxidase	ACCGGTCTTCTC T GAACAAACTTTATCCGGCGATCTTACCGTTTAG
precursor	CCGCTTTTAGCCCGGTGGGTCCTCCCA
Arabidopsis thaliana	TGGGAGGACCCACCGGGCTAAAAGCGGGTAAACGGTAAGATCGC	990
Arg7Term	GGGATAAAGTTTGTTC A GAGAAGACGGGTTGCCATAAAATTCAAA
CGA-TGA	GATTTTGTTTTTGAAAACGAAAACAAAAAAAA
	GTCTTCTC T GAACAAAC	991
	GTTTGTTC A GAGAAGAC	992

Freezing Tolerance	TCAAAAACAAAATCTTTGAATTTTATGGCAACCCGTCTTCTCAGAA	993
proline oxidase	CAAACTTTATCCGG T GATCTTACCGTTTACCGGCTTTTAGCCCGGT
precursor	GGGTCCTCCCACCGTGACTGCTTCCACCG
Arabidopsis thaliana	CGGTGGAAGCAGTCACGGTGGGAGGACCCAGCGGGCTAAAAGC	994
Arg13Term	GGGTAAACGGTAAGATC A CCGGATAAAGTTTGTTCTGAGAAGACG
CGA-TGA	GGTTGCCATAAAATTCAAAGATTTTGTTTTTGA
	TTATCCGG T GATCTTAC	995
	GTAAGATC A CCGGATAA	996

Freezing Tolerance	AAAATCTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTT	997
praline oxidase	ATCCGGCGATCTTA G CGTTTACCCGCTTTTAGCCCGGTGGGTCCT
precursor	CCCACCGTGACTGCTTCCACCGCCGTCGTC
Arabidopsis thaliana	GACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCACCGGG	998
Tyr15Term	CTAAAAGCGGGTAAACG C TAAGATCGCCGGATAAAGTTTGTTCGG
TAG-TAG	AGAAGAGGGGTTGCCATAAAATTCAAAGATTTT
	CGATCTTA G CGTTTACC	999
	GGTAAACG C TAAGATCG	1000

Freezing Tolerance	CTTTGAATTTTATGGCAACCCGTCTTCTCCGAACAAACTTTATCCG	1001
praline oxidase	GCGATCTTACCGTT A ACCCGCTTTTAGCCCGGTGGGTCCTCCCAC
precursor	CGTGACTGCTTCCACCGCCGTCGTCCCGGA
Arabidopsis thaliana	TCCGGGACGACGGCGGTGGAAGCAGTCACGGTGGGAGGACCCA	1002
Leu17Term	CCGGGCTAAAAGCGGGT T AACGGTAAGATCGGCGGATAAAGTTT
TTA-TAA	GTTCGGAGAAGACGGGTTGCCATAAAATTCAAAG
	TTACCGTT A ACCCGCTT	1003
	AAGCGGGT T AACGGTAA	1004

Freezing Tolerance	CCGGTGGGTCCTCCCACCGTGACTGCTTCCAGCGCCGTGGTCCC	1005
proline oxidase	GGAGATTCTCTCCTTT T GACAACAAGCACCGGAACCACCTCTTCA
precursor	CCACCCAAAACCCACCGAGCAATCTCACGATG
Arabidopsis thaliana	CATCGTGAGATTGCTCGGTGGGTTTTGGGTGGTGAAGAGGTGGT	1006
Gly42Term	TCCGGTGCTTGTTGTC A AAAGGAGAGAATCTCCGGGACGACGGC
GGA-TGA	GGTGGAAGCAGTCACGGTGGGAGGACCCACCGG
	TCTCCTTT T GACAACAA	1007
	TTGTTGTC A AAAGGAGA	1008

Lead Tolerance	ACATGAAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCT	1009
cyclic nucleotide-	AAACTATGAATTTC T GACAAGAGAAGTTTGTAAGGTCAGTGTTCCA
regulated ion channel	GATTTGTCTCATTGAATTCTAAGTCGTGA
Arabidopsis thaliana	TCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCTTAC	1010
Arg4Term	AAACTTCTCTTGTC A GAAATTCATAGTTTGAGACTAATAAGATTCAA
CGA-TGA	TACAAACAGAGATTTCACTGCTTCATGT
	TGAATTTC T GACAAGAG	1011
	CTCTTGTC A GAAATTCA	1012

Lead Tolerance	TGAAGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAA	1013
cyclic nucleotide-	CTATGAATTTCCGA T AAGAGAAGTTTGTAAGGTCAGTGTTCCAGAT
regulated ion channel	TTGTCTCATTGAATTCTAAGTCGTGAAGC
Arabidopsis thaliana	GCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGACCT	1014
Gln5Term	TACAAACTTCTCTT A TCGGAAATTCATAGTTTGAGACTAATAAGATT
CAA-TAA	CAATACAAACAGAGATTTCACTGCTTCA
	ATTTCCGA T AAGAGAAG	1015
	CTTCTCTT A TCGGAAAT	1016

Lead Tolerance	AGCAGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTAT	1017
cyclic nucleotide-	GAATTTCCGACAA T AGAAGTTTGTAAGGTCAGTGTTCCAGATTTGT
regulated ion channel	CTCATTGAATTCTAAGTCGTGAAGCTTA
Arabidopsis thaliana	TAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACACTGA	1018
Glu6Term	CCTTACAAACTTCT A TTGTCGGAAATTCATAGTTTGAGACTAATAA
GAG-TAG	GATTCAATACAAACAGAGATTTCACTGCT
	TCCGACAA T AGAAGTTT	1019
	AAACTTCT A TTGTCGGA	1020

Lead Tolerance	AGTGAAATCTCTGTTTGTATTGAATCTTATTAGTCTCAAACTATGAA	1021
cyclic nucleotide-	TTTCCGACAAGAG T AGTTTGTAAGGTCAGTGTTCCAGATTTGTCTC
regulated ion channel	ATTGAATTCTAAGTCGTGAAGCTTAATT
Arabidopsis thaliana	AATTAAGCTTCACGACTTAGAATTCAATGAGACAAATCTGGAACAC	1022
Lys7Term	TGACCTTACAAACT A CTCTTGTCGGAAATTCATAGTTTGAGACTAA
AAG-TAG	TAAGATTCAATACAAACAGAGATTTCACT
	GACAAGAG T AGTTTGTA	1023
	TACAAACT A CTCTTGTC	1024

Lead Tolerance	CATTGAATTCTAAGTCGTGAAGCTTAATTCGATTCTTCTTCACTTTC	1025
cyclic nucleotide-	TCGGATCAGGTTT T AAGATTGGAAGTCGGATAAGACTTCCTCCGA
regulated ion channel	CGTGGAATATTCCGGTAAAAACGAGATTC
Arabidopsis thaliana	GAATCTCGTTTTTACCGGAATATTCCACGTCGGAGGAAGTCTTATC	1026
Gln12Term	CGACTTCCAATCTT A AAACCTGATCCGAGAAAGTGAAGAAGAATC
CAA-TAA	GAATTAAGCTTCACGACTTAGAATTCAATG
	TCAGGTTT T AAGATTGG	1027
	CCAATCTT A AAACCTGA	1028

Lead Tolerance	TGGAAGTCAATCCCCCACGTTGAGCAGGTTGATGCATTGGGTAAA	1029
cyclic nucleotide-	GTTATGAATCACCGC T AAGACGAGTTTGTGAGGTTTCAGGATTGG
gated calmodulin-	AAATCAGAGAGAAGCTCTGAGGGAAATTTTC
binding ion channel	GAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTGAAACCT	1030
(CBP4)	CACAAACTCGTCTT A GCGGTGATTCATAACTTTAGCCAATGCATCA
Nicotiana Tabacum	ACCTGCTCAACGTGGGGGATTGACTTCCA
Gln5Term	ATCACCGC T AAGACGAG	1031
CAA-TAA	CTCGTCTT A GCGGTGAT	1032

Lead Tolerance	TCAATCCCCCACGTTGAGCAGGTTGATGCATTGGCTAAAGTTATG	1033
cyclic nucleotide-	AATCACCGCCAAGAC T AGTTTGTGAGGTTTCAGGATTGGAAATCA
gated calmodulin-	GAGAGAAGCTCTGAGGGAAATTTTCATGCTA
binding ion channel	TAGCATGAAAATTTCCCTCAGAGCTTCTCTCTGATTTCCAATCCTG	1034
(CBP4)	AAACCTCACAAACT A GTCTTGGCGGTGATTCATAACTTTAGCCAAT
Nicotiana Tabacum	GCATCAACCTGCTCAACGTGGGGGATTGA
Gly7Term	GCCAAGAC T AGTTTGTG	1035
GAG-TAG	CACAAACT A GTCTTGGC	1036

Lead Tolerance	GAGCAGGTTGATGCATTGGCTAAAGTTATGAATCACCGCCAAGAC	1037
cyclic nucleotide-	GAGTTTGTGAGGTTT T AGGATTGGAAATCAGAGAGAAGCTCTGAG
gated calmodulin-	GGAAATTTTCATGCTAAAGGTGGAGTCCACC
binding ion channel	GGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAGCTTCTCTC	1038
(CBP4)	TGATTTCCAATCCT A AAACCTCACAAACTCGTCTTGGCGGTGATTC
Nicotiana Tabacum	ATAACTTTAGCCAATGCATCAACCTGCTC
Gln12Term	TGAGGTTT T AGGATTGG	1039
CAG-TAG	CCAATCCT A AAACCTCA	1040

Lead Tolerance	TGATGCATTGGCTAAAGTTATGAATCACCGCCAAGACGAGTTTGT	1041
cyclic nucleotide-	GAGGTTTCAGGATTG T AAATCAGAGAGAAGCTCTGAGGGAAATTT
gated calmodulin-	TCATGCTAAAGGTGGAGTCCACCGAAGTAAA
binding ion channel	TTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGAG	1042
(CBP4)	CTTCTCTCTGATTT A CAATCCTGAAACCTCACAAACTCGTCTTGGC
Nicotiana Tabacum	GGTGATTCATAACTTTAGCCAATGCATCA
Trp14Term	CAGGATTG T AAATCAGA	1043
TGG-TGA	TCTGATTT A CAATCCTG	1044

Lead Tolerance	GATGCATTGGCTAAAGTTATGAATCACCGCCAAGACGAGTTTGTG	1045
cyclic nucleotide-	AGGTTTCAGGATTGG T AATCAGAGAGAAGCTGTGAGGGAAATTTT
gated calmoduin-	CATGCTAAAGGTGGAGTCCACCGAAGTAAAG
binding ion channel	CTTTACTTCGGTGGACTCCACCTTTAGCATGAAAATTTCCCTCAGA	1046
(CBP4)	GCTTCTCTCTGATT A CCAATCCTGAAACCTCACAAACTCGTCTTGG
Nicotiana Tabacum	CGGTGATTCATAACTTTAGCCAATGCATC
Lys15Term	AGGATTGG T AATCAGAG	1047
AAA-TAA	CTCTGATT A CCAATCCT	1048

Lead Tolerance	CTTGAAGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGG	1049
calmoduin binding	TGGAGATAATGATG T AAAGAGAGGACAGATATGTTAGATTTCAGG
transport protein	ACTGCAAATCAGAGCAATCTGTTATCTCAG
Hordeum vulgare	CTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATGTAACATA	1050
Glu2Term	TCTGTCCTCTCTTT A CATCATTATCTCCACCAGGCGAACAGTTAGC
GAA-TAA	AGCTAAGAGTGGTAGATCAATTCTTCAAG
	TAATGATG T AAAGAGAG	1051
	CTCTCTTT A CATCATTA	1052

Lead Tolerance	GAAGAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTG	1053
calmodulin binding	GAGATAATGATGGAA T GAGAGGACAGATATGTTAGATTTCAGGAC
transport protein	TGCAAATCAGAGCAATCTGTTATCTCAGAGA
Hordeum vulgare	TCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCTAAC	1054
Arg3Term	ATATCTGTCCTCTC A TTCCATCATTATCTCCACCAGGCGAACAGTT
AGA-TGA	AGCAGCTAAGAGTGGTAGATCAATTCTTC
	TGATGGAA T GAGAGGAC	1055
	GTCCTCTC A TTCCATCA	1056

Lead Tolerance	GAATTGATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAG	1057
calmodulin binding	ATAATGATGGAAAGA T AGGACAGATATGTTAGATTTCAGGACTGC
transport protein	AAATCAGAGCAATCTGTTATCTCAGAGAACG
Hordeum vulgare	CGTTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTGAAATCT	1058
Glu4Term	AACATATCTGTCCT A TCTTTCCATCATTATCTCCACCAGGCGAACA
GAG-TAG	GTTAGCAGCTAAGAGTGGTAGATCAATTC
	TGGAAAGA T AGGACAGA	1059
	TCTGTCCT A TCTTTCCA	1060

Lead Tolerance	ATCTACCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATG	1061
calmodulin binding	ATGGAAAGAGAGGAC T GATATGTTAGATTTCAGGACTGCAAATCA
transport protein	GAGCAATCTGTTATCTCAGAGAACGCAGTTT
Hordeum vulgare	AAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTTGCAGTCCTG	1062
Arg6Term	AAATCTAACATATC A GTCCTCTCTTTCCATCATTATCTCCACCAGG
AGA-TGA	CGAACAGTTAGCAGCTAAGAGTGGTAGAT
	GAGAGGAC T GATATGTT	1063
	AACATATC A GTCCTCTC	1064

Lead Tolerance	CCACTCTTAGCTGCTAACTGTTCGCCTGGTGGAGATAATGATGGA	1065
calmodulin binding	AAGAGAGGACAGATA G GTTAGATTTCAGGAGTGCAAATCAGAGCA
transport protein	ATCTGTTATCTCAGAGAACGCAGTTTCACCA
Hordeum vulgare	TGGTGAAACTGCGTTCTCTGAGATAACAGATTGCTCTGATTTGCA	1066
Tyr7Term	GTCCTGAAATCTAAC C TATCTGTCCTCTCTTTCCATCATTATCTCCA
TAT-TAG	CCAGGCGAACAGTTAGCAGCTAAGAGTGG
	GACAGATA G GTTAGATT	1067
	AATCTAAC C TATCTGTC	1068

2,4-DB resistance	ATCCTTCTCTGAGAAAAAACAACAGATCCGAATTTTATCTTTAATCA	1069
3-ketoacyl-CoA	GCCGGAAAAAATG T AGAAAGCGATCGAGAGACAACGCGTTCTTCT
thiolase	TGAGCATCTCCGACCTTCTTCTTCTTCTT
Arabidopsis thaliana	AAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGTTGT	1070
Glu2Term	CTCTCGATCGCTTTCT A CATTTTTTCCGGCTGATTAAAGATAAAATT
GAG-TAG	CGGATCTGTTGTTTTTTCTCAGAGAAGGAT
	AAAAAATG T AGAAAGCG	1071
	CGCTTTCT A CATTTTTT	1072

2,4-DB resistance	CTTCTCTGAGAAAAAACAACAGATCCGAATTTTATCTTTAATCAGC	1073
3-ketoacyl-CoA	CGGAAAAAATGGAG T AAGCGATCGAGAGACAACGCGTTCTTCTTG
thiolase	AGCATCTCCGACCTTCTTCTTCTTCTTCGC
Arabidopsis thaliana	GCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGAAGAACGCGT	1074
Lys3Term	TGTCTCTCGATCGCTT A CTCCATTTTTTCCGGCTGATTAAAGATAA
AAA-TAA	AATTCGGATCTGTTGTTTTTTCTCAGAGAAG
	AAATGGAG T AAGCGATC	1075
	GATCGCTT A CTCCATTT	1076

2,4-DB resistance	GAAAAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAA	1077
3-ketoacyl-CoA	TGGAGAAAGCGATC T AGAGACAACGCGTTCTTCTTGAGCATCTCC
thiolase	GACCTTCTTCTTCTTCTTCGCACAATTACG
Arabidopsis thaliana	CGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGCTCAAGA	1078
Glu6Term	AGAACGCGTTGTCTCT A GATCGCTTTCTCCATTTTTTCCGGCTGAT
GAG-TAG	TAAAGATAAAATTCGGATCTGTTGTTTTTTC
	AAGCGATC T AGAGACAA	1079
	TTGTCTCT A GATCGCTT	1080

2,4-DB resistance	AAAACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGG	1081
3-ketoacyl-CoA	AGAAAGCGATCGAG T GACAACGCGTTCTTCTTGAGCATCTCCGAC
thiolase	CTTCTTCTTCTTCTTCGCACAATTACGAGG
Arabidopsis thaliana	CCTCGTAATTGTGGGAAGAAGAAGAAGAAGGTCGGAGATGCTCAA	1082
Arg7Term	GAAGAACGCGTTGTC A CTCGATCGCTTTCTCCATTTTTTCCGGCT
AGA-TGA	GATTAAAGATAAAATTCGGATCTGTTGTTTT
	CGATCGAG T GACAACGC	1083
	GCGTTGTC A CTCGATCG	1084

2,4-DB resistance	ACAACAGATCCGAATTTTATCTTTAATCAGCCGGAAAAAATGGAGA	1085
3-ketoacyl-CoA	AAGCGATCGAGAGA T AACGCGTTCTTCTTGAGCATCTCCGACCTT
thiolase	CTTCTTCTTCTTCGCACAATTACGAGGCTT
Arabidopsis thaliana	AAGCCTCGTAATTGTGCGAAGAAGAAGAAGAAGGTCGGAGATGC	1086
Gln8Term	TCAAGAAGAACGCGTT A TCTCTCGATCGCTTTCTCCATTTTTTCCG
CAA-TAA	GCTGATTAAAGATAAAATTCGGATCTGTTGT
	TCGAGAGA T AACGCGTT	1087
	AACGCGTT A TCTCTCGA	1088

2,4-DB resistance	GAGAGACAAAGAGTTCTTCTTGAACATCTCCGTCCTTCTTCTTCTT	1089
glyoxysomal beta-	CCTCTCACAGCTTT T AAGGCTCTCTCTCTGCTTCAGCTTGCTTGGC
ketoacyol-thiolase	TGGGGACAGTGCTGCGTATCAGAGGACCT
precursor	AGGTCGTCTGATACGCAGCACTGTCCCCAGCCAAGCAAGCTGAA	1090
Brassica napus	GCAGAGAGAGAGCCTT A AAAGCTGTGAGAGGAAGAAGAAGAAGG
Glu26Term	ACGGAGATGTTCAAGAAGAACTCTTTGTCTCTC
GAA-TAA	ACAGCTTT T AAGGCTCT	1091
	AGAGCCTT A AAAGCTGT	1092

2,4-DB resistance	TTGAACATCTCCGTCCTTCTTCTTCTTCCTCTCACAGCTTTGAAGG	1093
glyoxysomal beta-	CTCTCTCTCTGCTT G AGCTTGCTTGGCTGGGGACAGTGCTGCGTA
ketoacyol-thiolase	TCAGAGGACCTCTCTCTATGGAGATGATGT
precursor	ACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGCACTGTCC	1094
Brassica napus	CCAGCCAAGCAAGCT C AAGCAGAGAGAGAGCCTTCAAAGCTGTG
Ser32Term	AGAGGAAGAAGAAGAAGGACGGAGATGTTCAA
TCA-TGA	CTCTGCTT G AGCTTGCT	1095
	AGCAAGCT C AAGCAGAG	1096

2,4-DB resistance	TCTCCGTCCTTCTTCTTCTTCCTCTCACAGCTTTGAAGGCTCTCTC	1097
glyoxysomal beta-	TCTGCTTCAGCTTG A TTGGCTGGGGACAGTGCTGCGTATCAGAG
ketoacyol-thiolase	GACCTCTCTCTATGGAGATGATGTAGTCATT
precursor	AATGACTACATCATCTCCATAGAGAGAGGTCCTCTGATACGCAGC	1098
Brassica napus	ACTGTCCCCAGCCAA T CAAGCTGAAGCAGAGAGAGAGCCTTCAAA
Cys34Term	GCTGTGAGAGGAAGAAGAAGAAGGACGGAGA
TGC-TGA	TCAGCTTG A TTGGCTGG	1099
	CCAGCCAA T CAAGCTGA	1100

2,4-DB resistance	TCCGTCCTTCTTCTTGTTCCTCTCACAGCTTTGAAGGCTCTCTCTC	1101
glyoxysomal beta-	TGCTTCAGCTTGCT A GGCTGGGGACAGTGCTGCGTATCAGAGGA
ketoacyol-thiolase	CCTCTCTCTATGGAGATGATGTAGTCATTGT
precursor	ACAATGACTACATCATCTCCATAGAGAGAGGTCGTCTGATACGCA	1102
Brassica napus	GCACTGTCCCCAGCC T AGCAAGCTGAAGCAGAGAGAGAGCCTTC
Leu35Term	AAAGCTGTGAGAGGAAGAAGAAGAAGGACGGA
TTG-TAG	AGCTTGCT A GGCTGGGG	1103
	CCCCAGCC T AGCAAGCT	1104

2,4-DB resistance	TCACAGCTTTGAAGGCTCTCTCTCTGCTTCAGCTTGCTTGGCTGG	1105
glyoxysomal beta-	GGACAGTGCTGCGTA G CAGAGGACCTCTCTCTATGGAGATGATGT
ketoacyol-thiolase	AGTCATTGTTGCGGCACATAGGACTGCACTA
precursor	TAGTGCAGTCCTATGTGCCGCAACAATGACTACATCATCTCCATA	1106
Brassica napus	GAGAGAGGTCGTCTG C TACGCAGCACTGTCCCCAGCCAAGCAAG
Tyr42Term	CTGAAGCAGAGAGAGAGCCTTCAAAGCTGTGA
TAT-TAG	GCTGCGTA G CAGAGGAC	1107
	GTCCTCTG C TACGCAGC	1108

2,4-DB resistance	CAACAGACAGGAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATTC	1109
3-ketoacyl-CoA	TTCTTCTCACAATTA Gee GAGTCCGCTCTTGCCGCATCAGTATGTGCT
thiolase B	GCAGGGGATAGCGCCGCATATCATAGGGCT
Mangifera indica	AGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATGC	1110
Tyr25Term	GGCAAGAGCGGACTC C TAATTGTGAGAAGAAGAATTAGAAGGGC
TAC-TAG	GGAGATGCTGGAGCAACACTTGCTGTCTGTTG
	CACAATTA G GAGTCCGC	1111
	GCGGACTC C TAATTGTG	1112

2,4-DB resistance	AACAGACAGCAAGTGTTGCTCCAGCATCTCCGCCCTTCTAATTCTT	1113
3-ketoacyol-CoA	CTTCTCACAATTAC T AGTCCGCTCTTGCCGCATCAGTATGTGCTGC
thiolase B	AGGGGATAGCGCCGCATATCATAGGGCTT
Magnifera indica	AAGCCCTATGATATGCGGCGCTATCCCCTGCAGCACATACTGATG	1114
Glu26Term	CGGCAAGAGCGGACT A GTAATTGTGAGAAGAAGAATTAGAAGGG
GAG-TAG	CGGAGATGCTGGAGCAACACTTGCTGTCTGTT
	ACAATTAC T AGTCCGCT	1115
	AGCGGACT A GTAATTGT	1116

2,4-DB resistance	TCCAGCATCTCCGCCCTTCTAATTCTTCTTCTCACAATTACGAGTC	1117
3-ketoacy\to-CoA	CGCTCTTGCCGCAT G AGTATGTGCTGCAGGGGATAGCGCCGCAT
thioblase B	ATCATAGGGCTTCTGTTTATGGAGACGATGT
Mangifera indica	ACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGCGCTATCC	1118
Ser32Term	CCTGCAGCACATACT C ATGCGGCAAGAGCGGACTCGTAATTGTGA
TCA-TGA	GAAGAAGAATTAGAAGGGCGGAGATGCTGGA
	TGCCGCAT G AGTATGTG	1119
	CACATACT C ATGCGGCA	1120

2,4-DB resistance	TCTCCGCCCTTCTAATTCTTCTTCTCACAATTACGAGTCCGCTCTT	1121
3-ketoacyl-CoA	GCCGCATCAGTATG A GCTGCAGGGGATAGCGCCGGATATCATAG
thiolase B	GGCTTCTGTTTATGGAGACGATGTGGTGATT
Mangifera indica	AATCACCACATCGTCTCCATAAACAGAAGCCCTATGATATGCGGC	1122
Cys34Term	GCTATCCCCTGCAGC T CATACTGATGCGGCAAGAGCGGACTCGT
TGT-TGA	AATTGTGAGAAGAAGAATTAGAAGGGCGGAGA
	TCAGTATG A GCTGCAGG	1123
	CCTGCAGC T CATACTGA	1124

2,4-DB resistance	TCACAATTACGAGTCCGCTCTTGCCGCATCAGTATGTGCTGCAGG	1125
3-ketoacyl-CoA	GGATAGCGCCGCATA G CATAGGGCTTGTGTTTATGGAGACGATGT
thiolase B	GGTGATTGTGGCAGGTCATCGTACTGCACTT
Mangifera indica	AAGTGCAGTAGGATGAGCTGCCACAATCACCACATCGTCTCCATA	1126
Tyr42Term	AACAGAAGCCCTATG C TATGCGGCGCTATCCCCTGCAGCACATAC
TAT-TAG	TGATGCGGCAAGAGCGGACTCGTAATTGTGA
	GCCGCATA G CATAGGGC	1127
	GCCCTATG C TATGCGGC	1128

2,4-DB resistance	GAAGGCGATCAACAGGCAGAGCATTTTGCTACATCATCTCCGGCC	1129
3-ketoacyl-CoA	TTCTTCTTCCGCTTA G ACAAATGAATCTTCGCTCTCTGCATCGGTT
thiolase	TGTGCAGCTGGGGATAGTGCTTCGTATCAA
Cucumis sativus	TTGATACGAAGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG	1130
Tyr22Term	CGAAGATTCATTTGT C TAAGCGGAAGAAGAAGGCCGGAGATGATG
TAG-TAG	TAGCAAAATGCTCTGGCTGTTGATCGCCTTC
	TCCGCTTA G ACAAATGA	1131
	TCATTTGT C TAAGCGGA	1132

2,4-DB resistance	ATCAACAGGCAGAGCATTTTGCTACATCATCTCCGGCCTTCTTCTT	1133
3-ketoacyl-CoA	CCGCTTACACAAAT T AATCTTCGCTCTCTGCATCGGTTTGTGCAGC
thiolase	TGGGGATAGTGCTTCGTATCAAAGGACAT
Cucumis sativus	ATGTCCTTTGATACGAAGCAGTATCCCCAGCTGCACAAACCGATG	1134
Glu25Term	CAGAGAGCGAAGATT A ATTTGTGTAAGCGGAAGAAGAAGGCCGG
GAA-TAA	AGATGATGTAGCAAAATGCTCTGCCTGTTGAT
	ACACAAAT T AATCTTCG	1135
	CGAAGATT A ATTTGTGT	1136

2,4-DB resistance	GGCAGAGCATTTTGCTACATCATCTCCGGCCTTCTTCTTCCGCTTA	1137
3-ketoacyl-CoA	CACAAATGAATCTT A GCTCTCTGCATCGGTTTGTGCAGCTGGGGA
thiolase	TAGTGCTTCGTATCAAAGGACATCGGTGTT
Cucumis sativus	AACACCGATGTCCTTTGATACGAAGCACTATCCCCAGCTGCACAA	1138
Ser27Term	ACCGATGCAGAGAGC T AAGATTCATTTGTGTAAGCGGAAGAAGAA
TCG-TAG	GGCCGGAGATGATGTAGCAAAATGCTCTGCC
	TGAATCTT A GCTCTCTG	1139
	CAGAGAGC T AAGATTCA	1140

2,4-DB resistance	TGCTACATCATCTCCGGCCTTCTTCTTCCGCTTACACAAATGAATC	1141
3-ketoacyl-CoA	TTCGCTCTCTGCAT A GGTTTGTGCAGCTGGGGATAGTGCTTCGTA
thiolase	TCAAAGGACATCGGTGTTTGGAGATGATGT
Cucumis sativus	ACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCACTATCCC	1142
Ser31Term	CAGCTGCACAAACC T ATGCAGAGAGCGAAGATTCATTTGTGTAAG
TCG-TAG	CGGAAGAAGAAGGCCGGAGATGATGTAGCA
	CTCTGCAT A GGTTTGTG	1143
	CACAAACC T ATGCAGAG	1144

2,4-DB resistance	TCATCTCCGGCCTTCTTCTTCCGCTTACACAAATGAATCTTCGCTC	1145
3-ketoacyl-CoA	TCTGCATCGGTTTG A GCAGCTGGGGATAGTGCTTCGTATCAAAGG
thiolase	ACATCGGTGTTTGGAGATGATGTCGTGATT
Cucumis sativus	AATCACGACATCATCTCCAAACACCGATGTCCTTTGATACGAAGCA	1146
Cys33Term	CTATCCCCAGCTGC T CAAACCGATGCAGAGAGCGAAGATTCATTT
TGT-TGA	GTGTAAGCGGAAGAAGAAGGCCGGAGATGA
	TCGGTTTG A GCAGCTGG	1147
	CCAGCTGC T CAAACCGA	1148

2A-DB resistance	GAAGGCAATCAACAGGCAGAGCATTCTGCTACATCATCTCCGGCC	1149
3-ketoacyl-CoA	TTCATCTTCGGCTTA G ACCCATGAATCTTCGCTCTCTGCATCGGTT
thiolase	TGTGCAGCTGGGGATAGTGCGTCGTATCAA
Cucurbita sp.	TTGATACGACGCACTATCCCCAGCTGCACAAACCGATGCAGAGAG	1150
Tyr22Term	CGAAGATTCATGGCT C TAAGCCGAAGATGAAGGCCGGAGATGAT
TAT-TAG	GTAGCAGAATGCTCTGCCTGTTGATTGCCTTC
	TCGGCTTA G AGCCATGA	1151
	TCATGGCT C TAAGCCGA	1152

2,4-DB resistance	ATCAACAGGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTT	1153
3-ketoacyl-CoA	CGGCTTATAGCCAT T AATCTTCGCTCTCTGCATCGGTTTGTGCAGC
thiolase	TGGGGATAGTGCGTCGTATCAAAGAACGT
Cucurbita sp.	ACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAAACCGATG	1154
Glu25Term	CAGAGAGCGAAGATT A ATGGCTATAAGCCGAAGATGAAGGCCGG
GAA-TAA	AGATGATGTAGCAGAATGCTCTGCCTGTTGAT
	ATAGCCAT T AATCTTCG	1155
	CGAAGATT A ATGGCTAT	1156

2,4-DB resistance	GGCAGAGCATTCTGCTACATCATCTCCGGCCTTCATCTTCGGCTT	1157
3-ketoacyl-CoA	ATAGCCATGAATCTT A GCTCTCTGCATCGGTTTGTGCAGCTGGGG
thiolase	ATAGTGCGTCGTATCAAAGAACGTCGGTGTT
Cucurbita sp.	AACACCGACGTTCTTTGATACGACGCACTATCCCCAGCTGCACAA	1158
Ser27Term	ACCGATGCAGAGAGCTAAGATTCATGGCTATAAGCCGAAGATGAA
TCG-TAG	GGCCGGAGATGATGTAGCAGAATGCTCTGCC
	TGAATCTT A GCTCTCTG	1159
	CAGAGAGC T AAGATTCA	1160

2,4-DB resistance	TGCTACATCATCTCCGGCCTTCATCTTCGGCTTATAGCCATGAATC	1161
3-ketoacyl-CoA	TTCGCTCTCTGCAT A GGTTTGTGCAGCTGGGGATAGTGCGTCGTA
thiolase	TCAAAGAACGTCGGTGTTTGGAGATGATGT
Cucurbita sp.	ACATCATCTCCAAACACCGACGTTCTTTGATACGACGCACTATCCC	1162
Ser31Term	CAGCTGCACAAACC T ATGCAGAGAGCGAAGATTCATGGCTATAAG
TCG-TAG	CCGAAGATGAAGGCCGGAGATGATGTAGCA
	CTCTGCAT A GGTTTGTG	1163
	CACAAACC T ATGCAGAG	1164

2,4-DB resistance	TCATCTCCGGCCTTCATCTTCGGCTTATAGCCATGAATCTTCGCTC	1165
3-ketoacyl-CoA	TCTGCATCGGTTTG A GCAGCTGGGGATAGTGCGTCGTATCAAAGA
thiolase	ACGTCGGTGTTTGGAGATGATGTCGTGATA
Cucurbita sp.	TATCACGACATCATCTCCAAACACCGACGTTCTTTGATACGACGCA	1166
Cys33Term	CTATCCCCAGCTGC T CAAACCGATGCAGAGAGCGAAGATTCATGG
TGT-TGA	CTATAAGCCGAAGATGAAGGCCGGAGATGA
	TCGGTTTG A GCAGCTGG	1167
	CCAGCTGC T CAAACCGA	1168

2,4 DB resistance	TCATAGTCTCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTG	1169
Pex14	CTATGGCAACTCAT T AGCAAACGCAACCTCCTTCCGATTTTCCCGC
Arabidopsis thaliana	TCTTGCCGATGAAAATTCCCAGATTCCAG
Gln5Term	CTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGGAA	1170
CAG-TAG	GGAGGTTGCGTTTGCT A ATGAGTTGCCATAGCAGCTCACTAACCT
	TGGAAGAATCCAAGCGGCAAAAGAGACTATGA
	CAACTCAT T AGCAAACG	1171
	CGTTTGCT A ATGAGTTG	1172

2,4 DB resistance	TAGTCTCTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTA	1173
Pex14	TGGCAACTCATCAG T AAACGCAACCTCCTTCCGATTTTCCCGCTCT
Arabidopsis thaliana	TGCCGATGAAAATTCCCAGATTGCAGGTT
Gln6Term	AACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAAAATCGG	1174
CAA-TAA	AAGGAGGTTGCGTTT A CTGATGAGTTGCCATAGCAGCTCACTAAC
	CTTGGAAGAATCCAAGCGGCAAAAGAGACTA
	CTCATCAG T AAACGCAA	1175
	TTGCGTTT A CTGATGAG	1176

2,4 DB resistance	CTTTTGCCGCTTGGATTCTTCCAAGGTTAGTGAGCTGCTATGGCA	1177
Pex14	ACTCATCAGCAAACG T AACCTCCTTCCGATTTTCCCGCTCTTGCCG
Arabidopsis thaliana	ATGAAAATTCCGAGATTCCAGGTTCAATTT
Gln8Term	AAATTGAACCTGGAATCTGGGAATTTTCATCGGCAAGAGCGGGAA	1178
CAA-TAA	AATCGGAAGGAGGTT A CGTTTGCTGATGAGTTGCCATAGCAGCTC
	ACTAACCTTGGAAGAATCCAAGCGGCAAAAG
	AGCAAACG T AACCTCCT	1179
	AGGAGGTT A CGTTTGCT	1180

2,4 DB resistance	GCTGCTATGGCAACTGATGAGCAAACGCAACCTCCTTCCGATTTT	1181
Pex14	CCCGCTCTTGCCGAT T AAAATTCCCAGATTCCAGGTTCAATTTACA
Arabidopsis thaliana	CCTTCTAATCATTATTTCTTAATTTTTCTT
Glu19Term	AAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAATTGAACCTGG	1182
GAA-TAA	AATCTGGGAATTTT A ATCGGCAAGAGCGGGAAAATCGGAAGGAG
	GTTGCGTTTGCTGATGAGTTGCCATAGCAGC
	TTGCCGAT T AAAATTCC	1183
	GGAATTTT A ATCGGCAA	1184

2,4 DB resistance	GCAACTCATCAGCAAACGCAACCTCCTTCCGATTTTCCCGCTCTT	1185
Pex14	GCCGATGAAAATTCC T AGATTCCAGGTTCAATTTACACCTTCTAAT
Arabidopsis thaliana	CATTATTTCTTAATTTTTCTTTGGTGGATT
Gln22Term	AATCCACCAAAGAAAAATTAAGAAATAATGATTAGAAGGTGTAAAT	1186
CAG-TAG	TGAACCTGGAATCT A GGAATTTTCATCGGCAAGAGCGGGAAAATC
	GGAAGGAGGTTGCGTTTGCTGATGAGTTGC
	AAAATTCC T AGATTCCA	1187
	TGGAATCT A GGAATTTT	1188

EXAMPLE 8

Production of Albino Mutants for the Analysis of Photosynthetic Processes

Plant productivity is limited by resources available and the ability of plants to harness these resources. The conversion of light to chemical energy, which is then used to synthesize carbohydrates, fatty acids, sugars, amino acids and other compounds, requires a complex system which combines the light harvesting apparatus of pigments and proteins. The value of light energy to the plant can only be realized when it is efficiently converted into chemical energy by photosynthesis and fed into various biochemical processes. Significant effort has therefore been directed at studying photosynthetic processes in plants in order to improve productivity and/or the efficiency of photosynthesis. The analysis of the photosynthetic process is substantially aided by the ability to produce albino plants. [0132]

The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism.

TABLE 18


Oligonucleotides to produce albino plants

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

White leaves	TTCTTTCCTGTGAAATTATCTGCTCAAATCTTTGGTTCCTGACGGAG	1189
Immutans	ATGGCGGCGATTT G AGGCATCTCCTCTGGTACGTTGACGATTTCA
Arabidopsis thaliana	CGGCCTTTGGTTACTCTTCGACGCTCTAG
Ser5Term	CTAGAGCGTCGAAGAGTAACCAAAGGCCGTGAAATCGTCAACGTA	1190
TCA-TGA	CCAGAGGAGATGCCT C AAATCGCCGCCATCTCCGTCAGGAACCAA
	AGATTTGAGCAGATAATTTCACAGGAAAGAA
	GGCGATTT G AGGCATCT	1191
	AGATGCCT C AAATCGCC	1192

White leaves	GCTCAAATCTTTGGTTCCTGACGGAGATGGCGGCGATTTCAGGCA	1193
Immutans	TCTCCTCTGGTACGT A GACGATTTCACGGCCTTTGGTTACTCTTCG
Arabidopsis thaliana	ACGCTCTAGAGCCGCCGTTTCGTACAGCTC
Leu12Term	GAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAGAGTAACCAAA 1194
TTG-TAG	GGCCGTGAAATCGTC T ACGTACCAGAGGAGATGCCTGAAATCGCC
	GCCATCTCCGTCAGGAACCAAAGATTTGAGC
	TGGTACGT A GACGATTT	1195
	AAATCGTC T ACGTACCA	1196

White leaves	TTTGGTTCCTGACGGAGATGGCGGCGATTTCAGGCATCTCCTCTG	1197
Immutans	GTACGTTGACGATTTGACGGCCTTTGGTTACTCTTCGACGCTCTAG
Arabidopsis thaliana	AGCCGCCGTTTCGTACAGCTCCTCTCACCG
Ser15Term	CGGTGAGAGGAGCTGTACGAAACGGCGGCTCTAGAGCGTCGAAG	1198
TCA-TGA	AGTAACCAAAGGCCGT C AAATCGTCAACGTACCAGAGGAGATGCC
	TGAAATCGCCGCCATCTCCGTCAGGAACCAAA
	GACGATTT G ACGGCCTT	1199
	AAGGCCGT C AAATCGTC	1200

White leaves	GCGGCGATTTCAGGCATCTCCTCTGGTACGTTGACGATTTCACGG	1201
Immutans	CCTTTGGTTACTCTT T GACGCTCTAGAGCCGCCGTTTCGTACAGCT
Arabidopsis thaliana	CCTCTCACCGATTGCTTCATCATCTTCCTC
Arg22Term	GAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTGTACGAAACG	1202
CGA-TGA	GCGGCTCTAGAGCGTC A AAGAGTAACCAAAGGCCGTGAAATCGTC
	AACGTACCAGAGGAGATGCCTGAAATCGCCGC
	TTACTCTT T GACGCTCT	1203
	AGAGCGTC A AAGAGTAA	1204

White leaves	TCAGGCATCTCCTCTGGTACGTTGACGATTTCACGGCCTTTGGTTA	1205
Immutans	CTCTTCGACGCTCT T GAGCCGCCGTTTCGTACAGCTCCTCTCACC
Arabidopsis thaliana	GATTGCTTCATCATCTTCCTCTCTCTTCTC
Arg25Term	GAGAAGAGAGAGGAAGATGATGAAGCAATCGGTGAGAGGAGCTG	1206
AGA-TGA	TACGAAACGGCGGCTC A AGAGCGTCGAAGAGTAACCAAAGGCCG
	TGAAATCGTCAACGTACCAGAGGAGATGCCTGA
	GACGCTCT T GAGCCGCC	1207
	GGCGGCTC A AGAGCGTC	1208

White leaves	GATTCTTGTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATT	1209
Immutans	TCTGCTATGAGTTTT T GAACCTCAGTTTCTTCATATTCTTGTTTTAG
Lycopersicon	AGCTAGGAGTTTTGAGAAGTCATCAGTTT
esculentum	AAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAGAATATGA	1210
Gly11Term	AGAAACTGAGGTTC A AAAACTCATAGCAGAAATCGAAATCGCCATT
GGA-TGA	CTTGATCCTTCTTCCTTCCCACAAGAATC
	TGAGTTTT T GAACCTCA	1211
	TGAGGTTC A AAAACTCA	1212

White leaves	GTGGGAAGGAAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTA	1213
Immutans	TGAGTTTTGGAACCT G AGTTTCTTCATATTCTTGTTTTAGAGCTAGG
Lycopersicon	AGTTTTGAGAAGTCATCAGTTTTATGCAA
esculentum	TTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTCTAAAACAAG	1214
Ser13Term	AATATGAAGAAACT C AGGTTCCAAAACTCATAGCAGAAATCGAAAT
TCA-TGA	CGCCATTCTTGATCCTTCTTCCTTCCCAC
	TGGAACCT G AGTTTCTT	1215
	AAGAAACT C AGGTTCCA	1216

White leaves	AAGAAGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGG	1217
Immutans	AACCTCAGTTTCTTGATATTCTTGTTTTAGAGCTAGGAGTTTTGAGA
Lycopersicon	AGTCATCAGTTTTATGCAATTCCCAGAA
esculentum	TTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTAGCTC	1218
Ser16Term	TAAAACAAGAATAT C AAGAAACTGAGGTTCCAAAACTCATAGCAGA
TCA-TGA	AATCGAAATCGCCATTCTTGATCCTTCTT
	AGTTTCTT G ATATTCTT	1219
	AAGAATAT C AAGAAACT	1220

White leaves	AGGATCAAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACC	1221
Immutans	TCAGTTTCTTCATA G TCTTGTTTTAGAGCTAGGAGTTTTGAGAAGTC
Lycopersicon	ATCAGTTTTATGCAATTCCCAGAACCCA
esculentum	TGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAACTCCTA	1222
Tyr17Term	GCTCTAAAACAAGA C TATGAAGAAACTGAGGTTCCAAAACTCATAG
TAT-TAG	CAGAAATCGAAATCGCCATTCTTGATCCT
	TCTTCATA G TCTTGTTT	1223
	AAACAAGA C TATGAAGA	1224

White leaves	AAGAATGGCGATTTCGATTTCTGCTATGAGTTTTGGAACCTCAGTT	1225
Immutans	TCTTCATATTCTTG A TTTAGAGCTAGGAGTTTTGAGAAGTCATCAGT
Lycopersicon	TTTATGCAATTCCCAGAACCCATGTCGG
esculentum	CCGACATGGGTTCTGGGAATTGCATAAAACTGATGACTTCTCAAAA	1226
Cys19Term	CTCCTAGCTCTAAA T CAAGAATATGAAGAAACTGAGGTTCCAAAAC
TGT-TGA	TCATAGCAGAAATCGAAATCGCCATTCTT
	TATTCTTG A TTTAGAGC	1227
	GCTCTAAA T CAAGAATA	1228

White leaves	CGCGTCCGATAAAAAAATCAAGAATGGCGATTTCCATATCTGCTAT	1229
Immutans	GAGTTTTCGAACTT G AGTTTCTTCTTCATATTCAGCATTTTTGTGCA
Capsicum annuum	ATTCCAAGAACCCATTTTGTTTGAATTC
Ser13Term	GAATTCAAACAAAATGGGTTCTTGGAATTGCACAAAAATGCTGAAT	1230
TCA-TGA	ATGAAGAAGAAACT C AAGTTCGAAAACTCATAGCAGATATGGAAAT
	CGCCATTCTTGATTTTTTTATCGGACGCG
	TCGAACTT G AGTTTCTT	1231
	AAGAAACT C AAGTTCGA	1232

White leaves	AAAAATCAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAAC	1233
Immutans	TTCAGTTTCTTCTT G ATATTCAGCATTTTTGTGCAATTCCAAGAACC
Capsicum annuum	CATTTTGTTTGAATTCTCTATTTTCACT
Ser17Term	AGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAATTGCACA	1234
TCA-TGA	AAAATGCTGAATAT C AAGAAGAAACTGAAGTTCGAAAACTCATAGC
	AGATATGGAAATCGCCATTCTTGATTTTT
	TTCTTCTT G ATATTCAG	1235
	CTGAATAT C AAGAAGAA	1236

White leaves	CAAGAATGGCGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGT	1237
Immutans	TTCTTCTTCATATT G AGCATTTTTGTGCAATTCCAAGAACCCATTTT
Capsicum annuum	GTTTGAATTCTCTATTTTCACTTAGGAA
Ser19Term	TTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGTTCTTGGAAT	1238
TCA-TGA	TGCACAAAAATGCT C AATATGAAGAAGAAACTGAAGTTCGAAAACT
	CATAGCAGATATGGAAATCGCCATTCTTG
	TTCATATT G AGCATTTT	1239
	AAAATGCT C AATATGAA	1240

White leaves	CGATTTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCA	1241
Immutans	TATTCAGCATTTT A GTGCAATTCCAAGAACCCATTTTGTTTGAATTC
Capsicum annuum	TCTATTTTCACTTAGGAATTCTCATAG
Leu21Term	CTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAATGGGT	1242
TTG-TAG	TCTTGGAATTGCAC T AAAATGCTGAATATGAAGAAGAAACTGAAGT
	TCGAAAACTCATAGCAGATATGGAAATCG
	AGCATTTT A GTGCAATT	1243
	AATTGCAC T AAAATGCT	1244

White leaves	TTCCATATCTGCTATGAGTTTTCGAACTTCAGTTTCTTCTTCATATT	1245
Immutans	CAGCATTTTTGTG A AATTCCAAGAACCCATTTTGTTTGAATTCTCTA
Capsicum annuum	TTTTCACTTAGGAATTCTCATAGAACT
Cys22Term	AGTTCTATGAGAATTCCTAAGTGAAAATAGAGAATTCAAACAAAAT	1246
TGC-TGA	GGGTTCTTGGAATT T CACAAAAATGCTGAATATGAAGAAGAAACTG
	AAGTTCGAAAACTCATAGCAGATATGGAA
	TTTTTGTG A AATTCCAA	1247
	TTGGAATT T CACAAAAA	1248

White leaves	TTCGGCACGAGGGAGAAGGAGCAGACCGAGGTGGCCGTCGAGG	1249
Immutans	AGTCCTTCCCCTTCAGG T AGACGGCTCCTCCTGACGAGCCACTGG
Oryza sativa	TCACCGCCGAGGAGAGCTGGGTGGTTAAGCTCG
Glu22Term	CGAGCTTAACCACCCAGCTCTCCTCGGCGGTGACCAGTGGCTCGT	1250
GAG-TAG	CAGGAGGAGCCGTCT A CCTGAAGGGGAAGGACTCCTCGACGGCC
	ACCTCGGTCTGCTCCTTCTCCCTCGTGCCGAA
	CCTTCAGG T AGACGGCT	1251
	AGCCGTCT A CCTGAAGG	1252

White leaves	GAGCAGACCGAGGTGGCCGTCGAGGAGTCCTTCCCCTTCAGGGA	1253
Immutans	GACGGCTCCTCCTGAC T AGCCACTGGTCACCGCCGAGGAGAGCT
Oryza sativa	GGGTGGTTAAGCTCGAGCAGTCCGTGAACATTT
Glu28Term	AAATGTTCACGGACTGCTCGAGCTTAACCACCCAGCTCTCCTCGG	1254
CAG-TAG	CGGTGACCAGTGGCT A GTCAGGAGGAGCCGTCTCCCTGAAGGGG
	AAGGACTCCTCGACGGCCACCTCGGTCTGCTC
	CTCCTGAC T AGCCACTG	1255
	CAGTGGCT A GTCAGGAG	1256

White leaves	GTCGAGGAGTCCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGA	1257
Immutans	GCCACTGGTCACCGCC T AGGAGAGCTGGGTGGTTAAGCTCGAGC
Oryza sativa	AGTCCGTGAACATTTTCCTCACGGAGTCAGTCA
Glu34Term	TGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCTTAA	1258
GAG-TAG	CCACCCAGCTCTCCTAGGCGGTGACCAGTGGCTCGTCAGGAGGA
	GCCGTCTCCCTGAAGGGGAAGGACTCCTCGAC
	TCACCGCC T AGGAGAGC	1259
	GCTCTCCT A GGCGGTGA	1260

White leaves	GAGGAGTCCTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCC	1261
Immutans	ACTGGTCACCGCCGAG T AGAGCTGGGTGGTTAAGCTCGAGCAGT
Oryza sativa	CCGTGAACATTTTCCTCACGGAGTCAGTCATCA
Glu35Term	TGATGACTGACTCCGTGAGGAAAATGTTCACGGACTGCTCGAGCT	1262
GAG-TAG	TAACCACCCAGCTCT A CTCGGCGGTGACCAGTGGCTCGTCAGGA
	GGAGCCGTCTCCCTGAAGGGGAAGGACTCCTC
	CCGCCGAG T AGAGCTGG	1263
	CCAGCTCT A CTCGGCGG	1264

White leaves	CTTCCCCTTCAGGGAGACGGCTCCTCCTGACGAGCCACTGGTCAC	1265
Immutans	CGCCGAGGAGAGCTG A GTGGTTAAGCTCGAGCAGTCCGTGAACA
Oryza sativa	TTTTCCTCACGGAGTCAGTCATCACGATACTT
Trp37Term	AAGTATCGTGATGACTGACTCCGTGAGGAAAATGTTCACGGACTG	1266
TGG-TGA	CTCGAGCTTAACCAC T CAGCTCTCCTCGGCGGTGACCAGTGGCTC
	GTCAGGAGGAGCCGTCTCCCTGAAGGGGAAG
	GAGAGCTG A GTGGTTAA	1267
	TTAACCAC T CAGCTCTC	1268

White leaves	TCCGGAGGAGGAAGGGGGATTCGACGAGGAGCTCACCCTCGCCG	1269
Immutans	GCGAGGACGGCGACTGAGTCGTCAGATTCGAGCAGTCCTTCAAC
Triticum aestivum	GTATTCCTCACGGATACTGTCATCTTTATACTC
Trp22Term	GAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAAGGACTG	1270
TGG-TGA	CTCGAATCTGACGAC T CAGTCGCCGTCCTCGCCGGCGAGGGTGA
	GCTCCTCGTCGAATCCCCCTTCCTCCTCCGGA
	GGCGACTG A GTCGTCAG	1271
	CTGACGAC T CAGTCGCC	1272

White leaves	GAGGAAGGGGGATTCGACGAGGAGCTCACCCTCGCCGGCGAGG	1273
Immutans	ACGGCGACTGGGTCGTC T GATTCGAGCAGTCCTTCAACGTATTCC
Triticum aestivum	TCACGGATACTGTCATCTTTATACTCGATATTC
Arg25Term	GAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATACGTTGAA	1274
AGA-TGA	GGACTGCTCGAATC A GACGACCCAGTCGCCGTCCTCGCCGGCGA
	GGGTGAGCTCCTCGTCGAATCCCCCTTCCTC
	GGGTCGTC T GATTCGAG	1275
	CTCGAATC A GACGACCC	1276

White leaves	GGGGGATTCGACGAGGAGCTCACCCTCGCCGGCGAGGACGGCG	1277
Immutans	ACTGGGTCGTCAGATTCTAGCAGTCCTTCAACGTATTCCTCACGGA
Triticum aestivum	TACTGTCATCTTTATACTCGATATTCTGTATC
Glu21Term	GATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAATAC	1278
GAG-TAG	GTTGAAGGACTGCT A GAATCTGACGACCCAGTCGCCGTCCTCGCC
	GGCGAGGGTGAGCTCCTCGTCGAATCCCCC
	TCAGATTC T AGCAGTCC	1279
	GGACTGCT A GAATCTGA	1280

White leaves	GGATTCGACGAGGAGCTCACCCTCGCCGGCGAGGACGGCGACTG	1281
Immutans	GGTCGTCAGATTCGAG T AGTCCTTCAACGTATTCCTCACGGATACT
Triticum aestivum	GTCATCTTTATACTCGATATTCTGTATCGTG
Gln28Term	CACGATACAGAATATCGAGTATAAAGATGACAGTATCCGTGAGGAA	1282
CAG-TAG	TACGTTGAAGGACTACTCGAATCTGACGACCCAGTCGCCGTCCTC
	GCCGGCGAGGGTGAGCTCCTCGTCGAATCC
	GATTCGAG T AGTCCTTC	1283
	GAAGGACT A CTCGAATC	1284

White leaves	CGAGCAGTCCTTCAACGTATTCCTCACGGATACTGTCATCTTTATA	1285
Immutans	CTCGATATTCTGTA G CGTGACCGCGACTACGCAAGGTTCTTCGTG
Triticum aestivum	CTCGAGACCATCGCCAGGGTGCCCTATTTC
Tyr46Term	GAAATAGGGCACCCTGGCGATGGTCTCGAGCACGAAGAACCTTG	1286
TAT-TAG	CGTAGTCGCGGTCACGCTACAGAATATCGAGTATAAAGATGACAG
	TATCCGTGAGGAATACGTTGAAGGACTGCTCG
	ATTCTGTA G CGTGACCG	1287
	CGGTCACG C TACAGAAT	1288

EXAMPLE 9

Altering Amino Acid Content of Plants

Another aim of biotechnology is to generate plants, especially crop plants, with added value traits. An example of such a trait is improved nutritional quality in food crops. For example, lysine, tryptophan and threonine, which are essential amino acids in the diet of humans and many animals, are limiting nutrients in most cereal crops. Consequently, grain-based diets, such as those based on corn, barley, wheat, rice, maize, millet, sorghum, and the like, must be supplemented with more expensive synthetic amino acids or amino-acid-containing oilseed protein meals. Increasing the lysine content of these grains or of any of the feed component crops would result in significant added value. [0134]
Naturally occurring mutants of plants that have different levels of particular essential amino acids have been identified. However, these mutants are generally not the result of increased free amino acid, but are instead the result of shifts in the overall protein profile of the grain. For example, in maize, reduced levels of lysine-deficient endosperm proteins (prolamines) are complemented by elevated levels of more lysine-rich proteins (albumins, globulins and glutelins). While nutritionally superior, these mutants are associated with reduced yields and poor grain quality, limiting their agronomic usefulness. [0135]
An alternative approach is to generate plants with mutations that render key amino acid biosynthetic enzymes insensitive to feedback inhibition. Many such mutations are known and mutation results in increased free amino acid. The increased production can optionally be coupled to increased expression of an abundant storage protein comprising the chosen amino acid. Alternatively, a normally abundant protein can be engineered to contain more of the target amino acid. [0136]

The attached table discloses exemplary oligonucleotide base sequences which can be used to generate site-specific mutations that remove feedback inhibition in plant amino acid biosynthetic enzymes.

TABLE 19


Genome-Altering Oligos Conferring Amino Acid Overproduction

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Met Overproduction	TATCCTCCAGGATCTTAAGATTTCCTCCTAATTTCGTCCGTCAGCT	1289
CGS	GAGCATTAAAGCCC A TAGAAACTGTAGCAACATCGGTGTTGCACA
Arabidopsis thaliana	GATCGTGGCGGCTAAGTGGTCCAACAACCC
Arg77His	GGGTTGTTGGACCACTTAGCCGCCACGATCTGTGCAACACCGAT	1290
CGT-CAT	GTTGCTACAGTTTCTA T GGGCTTTAATGCTCAGCTGACGGACGAA
	ATTAGGAGGAAATCTTAAGATCCTGGAGGATA
	TAAAGCCC A TAGAAACT	1291
	AGTTTCTA T GGGCTTTA	1292

Met Overproduction	TCTTAAGATTTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGC	1293
CGS	CCGTAGAAACTGTA A CAACATCGGTGTTGCACAGATCGTGGCGG
Arabidopsis thaliana	CTAAGTGGTCCAACAACCCATCCTCCGCGTT
Ser81Asn	AACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCCACGATCTG	1294
AGC-AAC	TGCAACACCGATGTTG T TACAGTTTCTACGGGCTTTAATGCTCAGC
	TGACGGACGAAATTAGGAGGAAATCTTAAGA
	AAACTGTA A CAACATCG	1295
	CGATGTTG T TACAGTTT	1296

Met Overproduction	TTTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGCCCGTAGAA	1297
CGS	ACTGTAGCAACATC A GTGTTGCACAGATCGTGGCGGCTAAGTGGT
Arabidopsis thaliana	CCAACAACCCATCCTCCGCGTTACCTTCGG
Gly84Ser	CCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGCC	1298
GGT-AGT	ACGATCTGTGCAACAC T GATGTTGCTACAGTTTCTACGGGCTTTAA
	TGCTCAGCTGACGGACGAAATTAGGAGGAAA
	GCAACATC A GTGTTGCA	1299
	TGCAACAC T GATGTTGC	1300

Met Overproduction	TTCCTCCTAATTTCGTCCGTCAGCTGAGCATTAAAGCCCGTAGAAA	1301
CGS	CTGTAGCAACATCG A TGTTGCACAGATCGTGGCGGCTAAGTGGTC
Arabidopsis thaliana	CAACAACCCATCCTCCGCGTTACCTTCGGC
Gly84Asp	GCCGAAGGTAACGCGGAGGATGGGTTGTTGGACCACTTAGCCGC	1302
GGT-GAT	CACGATCTGTGCAACA T CGATGTTGCTACAGTTTCTACGGGCTTTA
	ATGCTCAGCTGACGGACGAAATTAGGAGGAA
	CAACATCG A TGTTGCAC	1303
	GTGCAACA T CGATGTTG	1304

Met Overproduction	TATCGTCACTCATCCTCCGCTTCCCTCCCAACTTCGTCCGCCAGC	1305
CGS	TCAGCACCAAGGCCC A CCGCAACTGCAGCAACATCGGCGTCGCG
Fragraria vesca	CAGATCGTCGCGGCTTCGTGGTCCAACAAAGA
Arg73His	TCTTTGTTGGACCACGAAGCCGCGACGATCTGCGCGACGCCGAT	1306
CGC-CAC	GTTGCTGCAGTTGCGG T GGGCCTTGGTGCTGAGCTGGCGGACGA
	AGTTGGGAGGGAAGCGGAGGATGAGTGACGATA
	CAAGGCCC A CCGCAACT	1307
	AGTTGCGG T GGGCCTTG	1308

Met Overproduction	TCCTCCGCTTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGG	1309
CGS	CCCGCCGCAACTGCA A CAACATCGGCGTCGCGCAGATCGTCGCG
Fragraria vesca	GCTTCGTGGTCCAACAAAGACTCCGACCTTTC
Ser77Asn	GAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGCGACGATCTG	1310
AGC-AAC	CGCGACGCCGATGTTG T TGCAGTTGCGGCGGGCCTTGGTGCTGA
	GCTGGCGGACGAAGTTGGGAGGGAAGCGGAGGA
	CAACTGCA A CAACATCG	1311
	CGATGTTG T TGCAGTTG	1312

Met Overproduction	TTCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGGCCCGCCG	1313
CGS	CAACTGCAGCAACATC A GCGTCGCGCAGATCGTCGCGGCTTCGT
Fragraria vesca	GGTCCAACAAAGACTCCGACCTTTCGGCGGTGC
Gly80Ser	GCACCGCCGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGCG	1314
GGC-AGC	ACGATCTGCGCGACGC T GATGTTGGTGCAGTTGCGGCGGGCCTT
	GGTGCTGAGCTGGCGGACGAAGTTGGGAGGGAA
	GCAACATC A GCGTCGCG	1315
	CGCGACGC T GATGTTGC	1316

Met Overproduction	TCCCTCCCAACTTCGTCCGCCAGCTCAGCACCAAGGCCCGCCGC	1317
CGS	AACTGCAGCAACATCG A CGTCGCGCAGATCGTCGCGGCTTCGTG
Fragraria vesca	GTCCAACAAAGACTCCGACCTTTCGGCGGTGCC
Gly80Asp	GGCACCGCCGAAAGGTCGGAGTCTTTGTTGGACCACGAAGCCGC	1318
GGC-GAC	GACGATCTGCGCGACG T CGATGTTGCTGCAGTTGCGGCGGGCCT
	TGGTGCTGAGCTGGCGGACGAAGTTGGGAGGGA
	CAACATCG A CGTCGCGC	1319
	GCGCGACG T CGATGTTG	1320

Met Overproduction	TCTCCTCCCTCATCCTCCGCTTCCCTCCCAACTTCCAGCGCCAGC	1321
CGS	TAAGCACCAAGGCG A GCCGCAACTGCAGCAACATCGGCGTCGCG
Glycine max	CAAATCGTCGCCGCTTCGTGGTCGAACAACAG
Arg68His	CTGTTGTTCGACCACGAAGCGGCGACGATTTGCGCGACGCCGAT	1322
CGC-CAC	GTTGCTGCAGTTGCGGC T CGCCTTGGTGCTTAGCTGGCGCTGGA
	AGTTGGGAGGGAAGCGGAGGATGAGGGAGGAGA
	CCAAGGCG A GCCGCAAC	1323
	GTTGCGGC T CGCCTTGG	1324

Met Overproduction	TCCTCCGCTTCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGG	1325
CGS	CGCGCCGCAACTGCA A CAACATCGGCGTCGCGCAAATCGTCGCC
Glycine max	GCTTCGTGGTCGAACAACAGCGACAACTCTCC
Ser72Asn	GGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCGACGATTTG	1326
AGC-AAC	CGCGACGCCGATGTTG T TGCAGTTGCGGCGCGCCTTGGTGCTTA
	GCTGGCGCTGGAAGTTGGGAGGGAAGCGGAGGA
	CAACTGCA A CAACATCG	1327
	CGATGTTG T TGCAGTTG	1328

Met Overproduction	TTCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGGCGCGCCG	1329
CGS	CAACTGCAGCAACATC A GCGTCGCGCAAATCGTCGCCGCTTCGT
Glycine max	GGTCGAACAACAGCGACAACTCTCCGGCCGCCG
Gly75Ser	CGGCGGCCGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGCG	1330
GGC-AGC	ACGATTTGCGCGACGC T GATGTTGCTGCAGTTGCGGCGCGCCTT
	GGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGAA
	GCAACATC A GCGTCGCG	1331
	CGCGACGC T GATGTTGC	1332

Met Overproduction	TCCCTCCCAACTTCCAGCGCCAGCTAAGCACCAAGGCGCGCCGC	1333
CGS	AACTGCAGCAACATCG A CGTCGCGCAAATCGTCGCCGCTTCGTG
Glycine max	GTCGAACAACAGCGACAACTCTCCGGCCGCCGG
Gly75Asp	CCGGCGGCGGGAGAGTTGTCGCTGTTGTTCGACCACGAAGCGGC	1334
GGC-GAC	GACGATTTGCGCGACG T CGATGTTGCTGCAGTTGCGGCGCGCCT
	TGGTGCTTAGCTGGCGCTGGAAGTTGGGAGGGA
	CAACATCG A CGTCGCGC	1335
	GCGCGACG T CGATGTTG	1336

Met Overproduction	TGTCTTCTCTGATTTTCAGGTTTCCTCCTAATTTCGTGAGGCAGCT	1337
CGS	AAGCATTAAGGCT CAC AGGAATTGCAGCAATATTGGCGTGGCTCA
Solanum tuberosum	AGTTGTGGCGGCTTCCTGGTCTAACAACCA
Arg70His	TGGTTGTTAGACCAGGAAGCCGCCACAACTTGAGCCACGCCAATA	1338
AGG-CAC	TTGCTGCAATTCCT GTG AGCCTTAATGCTTAGCTGCCTCACGAAAT
	TAGGAGGAAACCTGAAAATCAGAGAAGACA
	TAAGGCT CAC AGGAATT	1339
	AATTCCT GTG AGCCTTA	1340

Met Overproduction	TTTTCAGGTTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGC	1341
CGS	TAGGAGGAATTGCA A CAATATTGGCGTGGCTCAAGTTGTGGCGG
Solanum tuberosum	CTTCCTGGTCTAACAACCAAGCCGGTCCTGA
Ser74Asn	TCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGCCACAACTTG	1342
AGC-AAC	AGCCACGCCAATATTGTTGCAATTCCTCCTAGCCTTAATGCTTAGC
	TGCCTCACGAAATTAGGAGGAAACCTGAAAA
	GAATTGCA A CAATATTG	1343
	CAATATTG T TGCAATTC	1344

Met Overproduction	TTTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGAGG	1345
CGS	AATTGCAGCAATATT A GCGTGGCTCAAGTTGTGGCGGCTTCCTGG
Solanum tuberosum	TCTAACAACCAAGCCGGTCCTGAATTCACTC
Gly77Ser	GAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGCC	1346
GGC-AGC	ACAACTTGAGCCACGC T AATATTGCTGCAATTCCTCCTAGCCTTAA
	TGCTTAGCTGCCTCACGAAATTAGGAGGAAA
	GCAATATT A GCGTGGGT	1347
	AGCCACGC T AATATTGC	1348

Met Overproduction	TTCCTCCTAATTTCGTGAGGCAGCTAAGCATTAAGGCTAGGAGGA	1349
CGS	ATTGCAGCAATATTG A CGTGGCTCAAGTTGTGGCGGCTTCCTGGT
Solanum tuberosum	CTAACAACCAAGCCGGTCCTGAATTCACTCC
Gly77Asp	GGAGTGAATTCAGGACCGGCTTGGTTGTTAGACCAGGAAGCCGC	1350
GGC-GAC	CACAACTTGAGCCACG T CAATATTGCTGCAATTCCTCCTAGCCTTA
	ATGCTTAGCTGCCTCACGAAATTAGGAGGAA
	CAATATTG A CGTGGCTC	1351
	GAGCCACG T CAATATTG	1352

Met Overproduction	CTTCCTCTCTTATCCTTCGCTTTCCTCCCAACTTTGTCCGTCAGCT	1353
CGS	CAGCACCAAGGCTCGCC A CAACTGCAGCAACATTGGTGTCGCAC
Mesembryanthemum	AGGTCGTCGCTGCCTCCTGGTCCAACAACTC
crystallinum	GAGTTGTTGGACCAGGAGGCAGCGACGACCTGTGCGACACCAAT	1354
Arg73His	GTTGCTGCAGTTG T GGCGAGCCTTGGTGCTGAGCTGACGGACAA
CGC-CAC	AGTTGGGAGGAAAGCGAAGGATAAGAGAGGAAG
	GGCTCGCC A CAACTGCA	1355
	TGCAGTTG T GGCGAGCC	1356

Met Overproduction	TCCTTCGCTTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGG	1357
CGS	CTCGCCGCAACTGCAACAACATTGGTGTCGCACAGGTCGTCGCT
Mesembryanthemum	GCCTCCTGGTCCAACAACTCCGATGCCGGCGC
crystallinum	GCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAGCGACGACCT	1358
Ser77Asn	GTGCGACACCAATG T TGTTGCAGTTGCGGCGAGCCTTGGTGCTG
AGC-AAC	AGCTGACGGACAAAGTTGGGAGGAAAGCGAAGGA
	CAACTGCA A CAACATTG	1359
	CAATGTTG T TGCAGTTG	1360

Met Overproduction	TTTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGGCTCGCCGC	1361
CGS	AACTGCAGCAACATT A GTGTCGCACAGGTCGTCGCTGCCTCCTG
Mesembryanthemum	GTCCAACAACTCCGATGCCGGCGCCACCTCTT
crystallinum	AAGAGGTGGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAGC	1362
Gly80Ser	GACGACCTGTGCGACAC T AATGTTGCTGCAGTTGCGGCGAGCCT
GGT-AGT	TGGTGCTGAGCTGACGGACAAAGTTGGGAGGAAA
	GCAACATT A GTGTCGCA	1363
	TGCGACAC T AATGTTGC	1364

Met Overproduction	TTCCTCCCAACTTTGTCCGTCAGCTCAGCACCAAGGCTCGCCGCA	1365
CGS	ACTGCAGCAACATTG A TGTCGCACAGGTCGTCGCTGCCTCCTGGT
Mesembryanthemum	CCAACAACTCCGATGCCGGCGCCACCTCTTG
crystallinum	CAAGAGGTGGCGCCGGCATCGGAGTTGTTGGACCAGGAGGCAG	1366
Gly80Asp	CGACGACCTGTGCGACA T CAATGTTGCTGCAGTTGCGGCGAGCC
GGT-GAT	TTGGTGCTGAGCTGACGGACAAAGTTGGGAGGAA
	CAACATTG A TGTCGCAC	1367
	GTGCGACA T CAATGTTG	1368

Met Overproduction	CCTCTGCTACCATCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGC	1369
CGS	TTAGCACCAAGGCACACGGCAACTGCAGCAACATCGGCGTCGCG
Zea mays	CAGATCGTCGCCGCCGCGTGGTCCGACTGCCC
Arg41His	GGGCAGTCGGACCACGCGGCGGCGACGATCTGCGCGACGCCGA	1370
CGC-CAC	TGTTGCTGCAGTTGCGG T GTGCCTTGGTGCTAAGCTGGCGGACA
	AAGTTTGGCGGAAAGCGGAGGATGGTAGCAGAGG
	CAAGGCAC A CCGCAACT	1371
	AGTTGCGG T GTGCCTTG	1372

Met Overproduction	TCCTCCGCTTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGG	1373
CGS	CACGCCGCAACTGCA A CAACATCGGCGTCGCGCAGATCGTCGCC
Zea mays	GCCGCGTGGTCCGACTGCCCCGCCGCTCGCCC
Ser45Asn	GGGCGAGCGGCGGGGCAGTCGGACCACGCGGCGGCGACGATCT	1374
AGC-AAC	GCGCGACGCCGATGTTG T TGCAGTTGCGGCGTGCCTTGGTGCTA
	AGCTGGCGGACAAAGTTTGGCGGAAAGCGGAGGA
	CAACTGCA A CAACATCG	1375
	CGATGTTG T TGCAGTTG	1376

Met Overproduction	TTTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGGCACGCCGC	1377
CGS	AACTGCAGCAACATC A GCGTCGCGCAGATCGTCGCCGCCGCGTG
Zea mays	GTCCGACTGCCCCGCCGCTCGCCCCCACTTAG
Gly48Ser	CTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCGG	1378
GGC-AGC	CGACGATCTGCGCGACGC T GATGTTGCTGCAGTTGCGGCGTGCC
	TTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAAA
	GCAACATC A GCGTCGCG	1379
	CGCGACGC T GATGTTGC	1380

Met Overproduction	TTCCGCCAAACTTTGTCCGCCAGCTTAGCACCAAGGCACGCCGCA	1381
CGS	ACTGCAGCAACATCG A CGTCGCGCAGATCGTCGCCGCCGCGTGG
Zea mays	TCCGACTGCCCCGCCGCTCGCCCCCACTTAGG
Gly48Asp	CCTAAGTGGGGGCGAGCGGCGGGGCAGTCGGACCACGCGGCG	1382
GGC-GAC	GCGACGATCTGCGCGACG T CGATGTTGCTGCAGTTGCGGCGTGC
	CTTGGTGCTAAGCTGGCGGACAAAGTTTGGCGGAA
	CAACATCG A CGTCGCGG	1383
	GCGCGACG T CGATGTTG	1384

Met Overproduction	GTATGAATGATCTGTGGGTGAAACACTGTGGGATTAGTCATACAG	1385
TS	GAAGTTTCAAGGATCGTGGAATGACTGTTTTGGTTAGTCAAGTTAA
Arabidopsis thaliana	TCGTCTGAGAAAGATGAAACGACCTGTGGT
Leu205Arg	ACCACAGGTCGTTTCATCTTTCTCAGACGATTAACTTGACTAACCA	1386
CTT-CGT	AAACAGTCATTCCA C GATCCTTGAAACTTCCTGTATGACTAATCCC
	ACAGTGTTTCACCCACAGATCATTCATAC
	CAAGGATC G TGGAATGA	1387
	TCATTCCA C GATCCTTG	1388

Met Overproduction	GCATGACTGATTTGTGGGTCAAACACTGTGGGATTAGCCATACTG	1389
TS	GTAGTTTTAAGGATCGTGGGATGACTGTTTTGGTGAGTCAAGTTAA
Solanum tuberosum	TCGCTTGCGGAAAATGCATAAACCGGTTGT
Leu198Arg	ACAACCGGTTTATGCATTTTCCGCAAGCGATTAACTTGACTCACCA	1390
CTT-CGT	AAACAGTCATCCCACGATCCTTAAAACTACCAGTATGGCTAATCCC
	ACAGTGTTTGACCCACAAATCAGTCATGC
	TAAGGATC G TGGGATGA	1391
	TCATCCCA C GATCCTTA	1392

Lys Overproduction	TCATTGGGCACACAGTGAACTGCTTTGGCTCTAGAATCAAAGTGA	1393
DHPS	TAGGCAACACAGGAA A CAACTCAACCAGAGAAGCCGTCCACGCA
Zea mays	ACAGAACAGGGATTTGCTGTTGGCATGCATGC
Ser157Asn	GCATGCATGCCAACAGCAAATCCCTGTTCTGTTGCGTGGACGGCT	1394
AGC-AAC	TCTCTGGTTGAGTTG T TTCCTGTGTTGCCTATCACTTTGATTCTAG
	AGCCAAAGCAGTTCACTGTGTGCCCAATGA
	CACAGGAA A CAACTCAA	1395
	TTGAGTTG T TTCCTGTG	1396

Lys Overproduction	GCTCTAGAATCAAAGTGATAGGCAACACAGGAAGCAACTCAACCA	1397
DHPS	GAGAAGCCGTCCACG A AACAGAACAGGGATTTGCTGTTGGCATG
Zea mays	CATGCGGCTCTCCACATCAATCCTTACTACGG
Ala166Val	CCGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGC	1398
GCA-GAA	AAATCCCTGTTCTGTT T CGTGGACGGCTTCTCTGGTTGAGTTGCTT
	CCTGTGTTGCCTATCACTTTGATTCTAGAGC
	CGTCCACG A AACAGAAC	1399
	GTTCTGTT T CGTGGACG	1400

Lys Overproduction	GGCTCTAGAATCAAAGTGATAGGCAACACAGGAAGCAACTCAACC	1401
DHPS	AGAGAAGCCGTCCAC A CAACAGAACAGGGATTTGCTGTTGGCAT
Zea mays	GCATGCGGCTCTCCACATCAATCCTTACTACG
Ala166Thr	CGTAGTAAGGATTGATGTGGAGAGCCGCATGCATGCCAACAGCA	1402
GCA-ACA	AATCCCTGTTCTGTTG T GTGGACGGCTTCTCTGGTTGAGTTGCTTC
	CTGTGTTGCCTATCACTTTGATTCTAGAGCC
	CCGTCCAC A CAACAGAA	1403
	TTCTGTTG T GTGGACGG	1404

Lys Overproduction	TTATTGGGCATACAGTTAACTGCTTTGGCACTAAAATTAAAGTGGT	1405
DHPS	CGGCAACACAGGAA A TAACTCAACAAGGGAGGCTATTCACGCAAC
Oryza sativa	TGAGCAGGGATTCGCTGTAGGTATGCACGC
Ser24Asn	GCGTGCATACCTACAGCGAATCCCTGCTCAGTTGCGTGAATAGCC	1406
AGT-AAT	TCCCTTGTTGAGTTA T TTCCTGTGTTGCCGACCACTTTAATTTTAGT
	GCCAAAGCAGTTAACTGTATGCCCAATAA
	CACAGGAA A TAACTCAA	1407
	TTGAGTTA T TTCCTGTG	1408

Lys Overproduction	GCACTAAAATTAAAGTGGTCGGCAACACAGGAAGTAACTCAACAA	1409
DHPS	GGGAGGCTATTCACGTAACTGAGCAGGGATTCGCTGTAGGTATG
Oryza sativa	CACGCGGCTCTCCACATCAATCCTTACTACGG
Ala133Val	CCGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGC	1410
GCA-GTA	GAATCCCTGCTCAGTT A CGTGAATAGCCTCCCTTGTTGAGTTACTT
	CCTGTGTTGCCGACCACTTTAATTTTAGTGC
	TATTCACG T AACTGAGC	1411
	GCTCAGTT A CGTGAATA	1412

Lys Overproduction	GGCACTAAAATTAAAGTGGTCGGCAACACAGGAAGTAACTCAACA	1413
DHPS	AGGGAGGCTATTCACACAACTGAGCAGGGATTCGCTGTAGGTAT
Oryza sativa	GCACGCGGCTCTCCACATCAATCCTTACTACG
Ala133Thr	CGTAGTAAGGATTGATGTGGAGAGCCGCGTGCATACCTACAGCG	1414
GCA-ACA	AATCCCTGCTCAGTTG T GTGAATAGCCTCCCTTGTTGAGTTACTTC
	CTGTGTTGCCGACCACTTTAATTTTAGTGCC
	CTATTCAC A CAACTGAG	1415
	CTCAGTTG T GTGAATAG	1416

Lys Overproduction	TCATCGGGCATACTGTTAACTGCTTTGGAGCCAACATTAAAGTGAT	1417
DHPS 1	AGGCAACACGGGAA A TAACTCAACCAGAGAAGCTGTTCACGCGA
Triticum aestivum	CAGAGCAGGGATTTGCTGTTGGCATGCATGC
Ser65Asn	GCATGCATGCCAACAGCAAATCCCTGCTCTGTCGCGTGAACAGCT	1418
AGT-AAT	TCTCTGGTTGAGTTA T TTCCCGTGTTGCCTATCACTTTAATGTTGG
	CTCCAAAGCAGTTAACAGTATGCCCGATGA
	CACGGGAA A TAACTCAA	1419
	TTGAGTTA T TTCCCGTG	1420

Lys Overproduction	GAGCCAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACCA	1421
DHPS 1	GAGAAGCTGTTCACGTGACAGAGCAGGGATTTGCTGTTGGCATG
Triticum aestivum	CATGCAGCTCTTCATGTCAATCCTTACTACGG
Ala174Val	CCGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCA	1422
GCG-GTG	AATCCCTGCTCTGTC A CGTGAACAGCTTCTCTGGTTGAGTTACTTC
	CCGTGTTGCCTATCACTTTAATGTTGGCTC
	TGTTCACG T GACAGAGC	1423
	GCTCTGTC A CGTGAACA	1424

Lys Overproduction	GGAGCCAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACC	1425
DHPS 1	AGAGAAGCTGTTCAC A CGACAGAGCAGGGATTTGCTGTTGGCAT
Triticum aestivum	GCATGCAGCTCTTCATGTCAATCCTTACTACG
Ala174Thr	CGTAGTAAGGATTGACATGAAGAGCTGCATGCATGCCAACAGCAA	1426
GCG-ACG	ATCCCTGCTCTGTCG T GTGAACAGCTTCTCTGGTTGAGTTACTTCC
	CGTGTTGCCTATCACTTTAATGTTGGCTCC
	CTGTTCAC A GGACAGAG	1427
	CTCTGTCG T GTGAACAG	1428

Lys Overproduction	TCATCGGGCACACTGTTAACTGCTTTGGAACTAACATTAAAGTGAT	1429
DHPS 2	AGGCAACACGGGAA A TAACTCAACTAGAGAAGCGATTCACGCTTC
Triticum aestivum	AGAGCAGGGATTTGCTGTTGGCATGCATGC
Ser154Asn	GCATGCATGCCAACAGCAAATCCCTGCTCTGAAGCGTGAATCGCT	1430
AGT-AAT	TCTCTAGTTGAGTTA T TTCCCGTGTTGCCTATCACTTTAATGTTAGT
	TCCAAAGCAGTTAACAGTGTGCCCGATGA
	CACGGGAA A TAACTCAA	1431
	TTGAGTTA T TTCCCGTG	1432

Lys Overproduction	GAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACTA	1433
DHPS 2	GAGAAGCGATTCACGTTTCAGAGCAGGGATTTGCTGTTGGCATGC
Triticum aestivum	ATGCAGCTCTCCATGTCAATCCTTACTATGG
Ala163Val	CCATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCA	1434
GCT-GTT	AATCCCTGCTCTGAA A CGTGAATCGCTTCTCTAGTTGAGTTACTTC
	CCGTGTTGCCTATCACTTTAATGTTAGTTC
	GATTCACG T TTCAGAGC	1435
	GCTCTGAA A CGTGAATC	1436

Lys Overproduction	GGAACTAACATTAAAGTGATAGGCAACACGGGAAGTAACTCAACT	1437
DHPS 2	AGAGAAGCGATTCACACTTCAGAGCAGGGATTTGCTGTTGGCATG
Triticum aestivum	CATGCAGCTCTCCATGTCAATCCTTACTATG
Ala163Thr	CATAGTAAGGATTGACATGGAGAGCTGCATGCATGCCAACAGCAA	1438
GCT-ACT	ATCCCTGCTCTGAAG T GTGAATCGCTTCTCTAGTTGAGTTACTTCC
	CGTGTTGCCTATCACTTTAATGTTAGTTCC
	CGATTCAC A CTTCAGAG	1439
	CTCTGAAG T GTGAATCG	1440

Lys Overproduction	CTCATTGGGCATACTGTGAACTGCTTTGGCTCTAGAATTAAAGTGA	1441
DHPS	TAGGCAACACAGGAA A TAACTCAACCAGAGAAGCTGTTCACGCAA
Coix lacryma-jobi	CAGAGCAGGGATTTGCTGTTGGCATGCATG
Ser154Asn	CATGCATGCCAACAGCAAATCCCTGCTCTGTTGCGTGAACAGCTT	1442
AGT-AAT	CTCTGGTTGAGTTA T TTCCTGTGTTGCCTATCACTTTAATTCTAGA
	GCCAAAGCAGTTCACAGTATGCCCAATGAG
	CACAGGAA A TAACTCAA	1443
	TTGAGTTA T TTCCTGTG	1444

Lys Overproduction	GCTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAACCA	1445
DHPS	GAGAAGCTGTTCACG T AACAGAGCAGGGATTTGCTGTTGGCATGC
Coix lacryma-jobi	ATGCAGCTCTCCACATCAATCCTTACTATGG
Ala163Val	CCATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCA	1446
GCA-GTA	AATCCCTGCTCTGTT A CGTGAACAGCTTCTCTGGTTGAGTTACTTC
	CTGTGTTGCCTATCACTTTAATTCTAGAGC
	TGTTCACG T AACAGAGC	1447
	GCTCTGTT A CGTGAACA	1448

Lys Overproduction	GGCTCTAGAATTAAAGTGATAGGCAACACAGGAAGTAACTCAACC	1449
DHPS	AGAGAAGCTGTTCAC A CAACAGAGCAGGGATTTGCTGTTGGCATG
Coix lacryma-jobi	CATGCAGCTCTCCACATCAATCCTTACTATG
Ala163Thr	CATAGTAAGGATTGATGTGGAGAGCTGCATGCATGCCAACAGCAA	1450
GCA-ACA	ATCCCTGCTCTGTTG T GTGAACAGCTTCTCTGGTTGAGTTACTTCC
	TGTGTTGCCTATCACTTTAATTCTAGAGCC
	CTGTTCAC A CAACAGAG	1451
	CTCTGTTG T GTGAACAG	1452

Lys Overproduction	TCATTGGTCACACAGTCAATTGTTTTGGAGGGTCCATCAAAGTCAT	1453
DHPS	CGGGAACACTGGAA A CAACTCCACAAGGGAAGCAATCCATGCAA
Nicotiana tabacum	CTGAACAGGGATTTGCTGTAGGTATGCATGC
Ser136Asn	GCATGCATACCTACAGCAAATCCCTGTTCAGTTGCATGGATTGCTT	1454
AGC-AAC	CCCTTGTGGAGTTG T TTCCAGTGTTCCCGATGACTTTGATGGACC
	CTCCAAAACAATTGACTGTGTGACCAATGA
	CACTGGAA A CAACTCCA	1455
	TGGAGTTG T TTCCAGTG	1456

Lys Overproduction	GAGGGTCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCACAA	1457
DHPS	GGGAAGCAATCCATG T AACTGAACAGGGATTTGCTGTAGGTATGC
Nicotiana tabacum	ATGCAGCTCTTCACATTAATCCCTACTATGG
Ala145Val	CCATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCA	1458
GCA-GTA	AATCCCTGTTCAGTT A CATGGATTGCTTCCCTTGTGGAGTTGCTTC
	CAGTGTTCCCGATGACTTTGATGGACCCTC
	AATCCATG T AACTGAAC	1459
	GTTCAGTT A CATGGATT	1460

Lys Overproduction	GGAGGGTCCATCAAAGTCATCGGGAACACTGGAAGCAACTCCAC	1461
DHPS	AAGGGAAGCAATCCAT A CAACTGAACAGGGATTTGCTGTAGGTAT
Nicotiana tabacum	GCATGCAGCTCTTCACATTAATCCCTACTATG
Ala145Thr	CATAGTAGGGATTAATGTGAAGAGCTGCATGCATACCTACAGCAA	1462
GCA-ACA	ATCCCTGTTCAGTTG T ATGGATTGCTTCCCTTGTGGAGTTGCTTCC
	AGTGTTCCCGATGACTTTGATGGACCCTCC
	CAATCCAT A CAACTGAA	1463
	TTCAGTTG T ATGGATTG	1464

Lys Overproduction	TTATAGGCCATACCGTTAACTGTTTTGGCGGAAGCATCAAAGTCAT	1465
DHPS	TGGAAACACTGGAA A CAATTCGACTAGAGAAGCAATCCACGCGAC
Arabidopsis thaliana	TGAACAAGGATTCGCGGTTGGAATGCATGC
Ser142Asn	GCATGCATTCCAACCGCGAATCCTTGTTCAGTCGCGTGGATTGCT	1466
AGC-AAC	TCTCTAGTCGAATTG T TTCCAGTGTTTCCAATGACTTTGATGCTTC
	CGCCAAAACAGTTAACGGTATGGCCTATAA
	CACTGGAA A CAATTCGA	1467
	TCGAATTG T TTCCAGTG	1468

Lys Overproduction	GCGGAAGCATCAAAGTCATTGGAAACACTGGAAGCAATTCGACTA	1469
DHPS	GAGAAGCAATCCACG T GACTGAACAAGGATTCGCGGTTGGAATG
Arabidopsis thaliana	CATGCTGCTCTTCATATAAACCCTTACTATGG
Ala151Val	CCATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCG	1470
GCG-GTG	AATCCTTGTTCAGTC A CGTGGATTGCTTCTCTAGTCGAATTGCTTC
	CAGTGTTTCCAATGACTTTGATGCTTCCGC
	AATCCACG T GACTGAAC	1471
	GTTCAGTC A CGTGGATT	1472

Lys Overproduction	GGCGGAAGCATCAAAGTCATTGGAAACACTGGAAGCAATTCGACT	1473
DHPS	AGAGAAGCAATCCAC A CGACTGAACAAGGATTCGCGGTTGGAAT
Arabidopsis thaliana	GCATGCTGCTCTTCATATAAACCCTTACTATG
Ala151Thr	CATAGTAAGGGTTTATATGAAGAGCAGCATGCATTCCAACCGCGA	1474
GCG-ACG	ATCCTTGTTCAGTCG T GTGGATTGCTTCTCTAGTCGAATTGCTTCC
	AGTGTTTCCAATGACTTTGATGCTTCCGCC
	CAATCCAC A CGACTGAA	1475
	TTCAGTCG T GTGGATTG	1476

Lys Overproduction	TTATTGCTCATACAGTCAACTGTTTTGGTGGGAAAATTAAGGTTAT	1477
DHPS	TGGAAATACTGGAA A CAACTCCACCAGGGAAGCAATTCATGCCAC
Glycine max	TGAGCAGGGTTTTGCTGTTGGAATGCATGC
Ser103Asn	GCATGCATTCCAACAGCAAAACCCTGCTCAGTGGCATGAATTGCT	1478
AGC-AAC	TCCCTGGTGGAGTTGTTTCCAGTATTTCCAATAACCTTAATTTTCC
	CACCAAAACAGTTGACTGTATGAGCAATAA
	TACTGGAA A CAACTCCA	1479
	TGGAGTTG T TTCCAGTA	1480

Lys Overproduction	GTGGGAAAATTAAGGTTATTGGAAATACTGGAAGCAACTCCACCA	1481
DHPS	GGGAAGCAATTCATG T CACTGAGCAGGGTTTTGCTGTTGGAATGC
Glycine max	ATGCTGCCCTTCACATAAACCCTTACTATGG
Ala112Val	CCATAGTAAGGGTTTATGTGAAGGGCAGCATGCATTCCAACAGCA	1482
GCC-GTC	AAACCCTGCTCAGTG A CATGAATTGCTTCCCTGGTGGAGTTGCTT
	CCAGTATTTCCAATAACCTTAATTTTCCCAC
	AATTCATG T CACTGAGC	1483
	GCTCAGTG A CATGAATT	1484

Lys Overproduction	GGTGGGAAAATTAAGGTTATTGGAAATACTGGAAGCAACTCCACC	1485
DHPS	AGGGAAGCAATTCAT A CCACTGAGCAGGGTTTTGCTGTTGGAATG
Glycine max	CATGCTGCCCTTCACATAAACCCTTACTATG
Ala112Thr	CATAGTAAGGGTTTATGTGAAGGGCAGCATGGATTCCAACAGCAA	1486
GCC-ACC	AACCCTGCTCAGTGG T ATGAATTGCTTCCCTGGTGGAGTTGCTTC
	CAGTATTTCCAATAACCTTAATTTTCCCACC
	CAATTCAT A CCACTGAG	1487
	CTCAGTGG T ATGAATTG	1488

Trp Overproduction	CTTGCAGGAGACATATTTCAGATCGTGCTGAGTCAACGTTTTGAG	1489
AS	CGGCGAACATTTGCA A ACCCCTTTGAAGTTTATAGAGCACTAAGA
Arabidopsis thaliana	GTTGTGAATCCAAGTCCGTATATGGGTTATT
Asp341Asn	AATAACCCATATACGGACTTGGATTCACAACTCTTAGTGCTCTATA	1490
GAG-AAC	AACTTCAAAGGGGT T TGCAAATGTTCGCCGCTCAAAACGTTGACT
	CAGCACGATCTGAAATATGTCTCCTGCAAG
	CATTTGCA A ACCCCTTT	1491
	AAAGGGGT T TGCAAATG	1492

Trp Overproduction	GCTGCAGGAGACATATTTCAAATCGTTTTAAGTCAACGCTTTGAGA	1493
AS	GAAGAACATTTGCT A ACCCATTTGAAGTGTACAGAGCATTAAGAAT
Nicotiana tabacum	TGTGAATCCAAGCCCATATATGACTTACA
Asp326Asn	TGTAAGTCATATATGGGCTTGGATTCACAATTCTTAATGCTCTGTA	1494
GAC-AAC	CACTTCAAATGGGT T AGCAAATGTTCTTCTCTCAAAGCGTTGACTT
	AAAACGATTTGAAATATGTCTCCTGCAGC
	CATTTGCT A ACCCATTT	1495
	AAATGGGT T AGCAAATG	1496

Trp Overproduction	CTAGCTGGTGACATTTTTCAAGTAGTCTTAAGCCAGCGTTTTGAGA	1497
AS	GGCGTACATTTGCT A ACCCCTTTGAGGTGTACCGTGCATTGCGTA
Oryza sativa	TTGTCAATCCTAGTCCTTATATGGCCTATC
Asp323Asn	GATAGGCCATATAAGGACTAGGATTGACAATACGCAATGCACGGT	1498
GAC-AAC	ACACCTCAAAGGGGT T AGCAAATGTACGCCTCTCAAAACGCTGGC
	TTAAGACTACTTGAAAAATGTCACCAGCTAG
	CATTTGCT A ACCCCTTT	1499
	AAAGGGGT T AGCAAATG	1500

Trp Overproduction	CTTGCTGGTGACATATTCCAGATCGTACTAAGTCAGCGTTTTGAAA	1501
AS	GGCGAACGTTCGCA A ACCCATTTGAAATCTATAGATCACTGAGGA
Ruta graveolens	TTGTTAATCCAAGCCCATATATGACTTATT
Asp354Asn	AATAAGTCATATATGGGCTTGGATTAACAATCCTCAGTGATCTATA	1502
GAC-AAC	GATTTCAAATGGGT T TGCGAACGTTCGCCTTTCAAAACGCTGACTT
	AGTACGATCTGGAATATGTCACCAGCAAG
	CGTTCGCA A ACCCATTT	1503
	AAATGGGT T TGCGAACG	1504

Trp Overproduction	CTGGCTGGGGACATATTCCAGCTTGTCCTAAGTCAGCGTTTTGAA	1505
AS	CGGCGAACATTTGCA A ATCCATTTGAAGTCTACCGAGCATTGAGA
Catharanthus roseus	ATTGTCAACCCAAGTCCATATATGACTTATT
Asp354Asn	AATAAGTCATATATGGACTTGGGTTGACAATTCTCAATGCTCGGTA	1506
GAT-AAT	GACTTCAAATGGAT T TGCAAATGTTCGCCGTTCAAAACGCTGACTT
	AGGACAAGCTGGAATATGTCCCCAGCCAG
	CATTTGCA A ATCCATTT	1507
	AAATGGAT T TGCAAATG	1508

EXAMPLE 10

Production of Modified Starch in Plants

A principal aim of biotechnology is the improvement of crop plants for food value, agriculture, and to produce a range of plant-derived raw materials. Along with oils, fats and proteins, polysaccharides constitute the main raw materials derived from plants, and apart from cellulose, the storage polymer starch is the most important polysaccharide raw material. Starch is derived from a range of plants, but maize is the most important cultivated plant for the production of starch. [0138]
The polysaccharide starch is a polymer made up of glucose molecules. However, starch is not a homogeneous raw material and is, in fact, a highly complex mixture of various types of molecules which differ from each other, for example, in their degree of polymerization and in the degree of branching of the glucose chains. For example, amylose-starch is a basically non-branched polymer made up of α-1,4-glycosidically branched glucose molecules, and amylopectin-starch is a complex mixture of variously branched glucose chains. The branching results from additional α-1,6-glycosidic linkages. In plants from which starch is typically isolated, for example maize or potato, the starch is approximately 25% amylose-starch and 75% amylopectin-starch. [0139]
In maize, various mutants in starch metabolism are known, for example waxy, sugary, shrunken and opaque-2. In addition to producing a modified starch, these mutations greatly improve grain quality in maize, and thus expand the use of maize not only as the food but also for the important industrial materials in food chemistry. It would therefore be advantageous to be able readily to obtain mutants in these genes in particular maize genotypes as well as other plants. Such plants can be obtained, for example, using traditional breeding methods and through specific genetic modification by means of recombinant DNA techniques. [0140]

The attached tables disclose exemplary oligonucleotide base sequences which can be used to generate site-specific mutations in genes involved in starch metabolism.

TABLE 20


Genome-Altering Oligos Conferring Increased Starch

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Increased Starch	GAACTTGAGACTGAGAAAAGGGATCCAAGGACAGTTGCTTCCATT	1509
ADPGPP	ATTCTTGGAGGTGGA AA AGGAACTCGACTCTTTCCTCTCACAAAA
Arabidopsis thaliana	CGCCGCGCCAAGCCTGCCGTTCCTATCGGGG
Ala99Lys	CCCCGATAGGAACGGCAGGCTTGGCGCGGCGTTTTGTGAGAGGA	1510
GCA-AAA	AAGAGTCGAGTTCCT TT TCCACCTCCAAGAATAATGGAAGCAACT
	GTCCTTGGATCCCTTTTCTCAGTCTCAAGTTC
	GAGGTGGA AA AGGAACT	1511
	AGTTCCT TT TCCACCTC	1512

Increased Starch	CAAAACGCCGCGCCAAGCCTGCCGTTCCTATCGGGGGAGCCTAT	1513
ADPGPP	AGGTTGATAGATGTAC T AATGAGCAATTGTATTAACAGCGGAATCA
Arabidopsis thaliana	ACAAAGTCTACATACTCACACAATATAACTC
Pro127Leu	GAGTTATATTGTGTGAGTATGTAGACTTTGTTGATTCCGCTGTTAA	1514
CCA-CTA	TACAATTGCTCATT A GTACATCTATCAACCTATAGGCTCCCCCGAT
	AGGAACGGCAGGCTTGGCGCGGCGTTTTG
	AGATGTAC T AATGAGCA	1515
	TGCTCATT A GTACATCT	1516

Increased Starch	TCACACAATATAACTCAGCATCATTGAACAGGCATTTAGCCCGTGC	1517
ADPGPP	TTACAACTCCAAT AAT CTTGGCTTTGGAGATGGCTATGTTGAGGTT
Arabidopsis thaliana	CTTGCGGCCACTCAAACGCCAGGAGAATC
Gly162Asn	GATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA	1518
GGA-AAT	TCTCCAAAGCCAAG ATT ATTGGAGTTGTAAGCACGGGGTAAATGC
	CTGTTCAATGATGCTGAGTTATATTGTGTGA
	CTCCAAT AAT CTTGGCT	1519
	AGCCAAG ATT ATTGGAG	1520

Increased Starch	TCACACAATATAACTCAGCATCATTGAACAGGCATTTAGCCCGTGC	1521
ADPGPP	TTACAACTCCAAT AAC CTTGGCTTTGGAGATGGCTATGTTGAGGTT
Arabidopsis thaliana	CTTGCGGCCACTCAAACGCCAGGAGAATC
Gly162Asn	GATTCTCCTGGCGTTTGAGTGGCCGCAAGAACCTCAACATAGCCA	1522
GGA-AAC	TCTCCAAAGCCAAG GTT ATTGGAGTTGTAAGCACGGGCTAAATGC
	CTGTTCAATGATGCTGAGTTATATTGTGTGA
	CTCCAAT AAC CTTGGCT	1523
	AGCCAAG GTT ATTGGAG	1524

Increased Starch	GTTTGAGAGAAGAAAGGTAGACCCGCAAAATGTGGCTGCAATCAT	1525
ADPGPP	TCTAGGAGGAGGCAA A GGAGCTAAACTCTTCCCTCTTACAATGAG
Arabidopsis thaliana	AGCCGCAACACCAGCTGTAAATATTCATCTT
Asn100Lys	AAGATGAATATTTACAGCTGGTGTTGCGGCTCTCATTGTAAGAGG	1526
AAT-AAA	GAAGAGTTTAGCTCC T TTGCCTCCTCCTAGAATGATTGCAGCCAC
	ATTTTGCGGGTCTACCTTTCTTCTCTCAAAC
	GGAGGCAA A GGAGCTAA	1527
	TTAGCTCC T TTGCCTCC	1528

Increased Starch	CTTGTGTCTTCAAATTATGTTAGGTTCCTGTTGGTGGATGCTACAG	1529
ADPGPP	GCTGATCGATATCC T GATGAGTAACTGTATTAACAGCTGCATCAAC
Arabidopsis thaliana	AAGATATTTGTGCTGACACAGTTCAACTC
Pro128Leu	GAGTTGAACTGTGTCAGCACAAATATCTTGTTGATGCAGCTGTTAA	1530
CCG-CTG	TACAGTTACTCATC A GGATATCGATCAGCCTGTAGCATCCACCAA
	CAGGAACCTAACATAATTTGAAGACACAAG
	CGATATCC T GATGAGTA	1531
	TACTCATC A GGATATCG	1532

Increased Starch	TGACACAGTTCAACTCAGCTTCCCTTAATCGACATTTAGCACGAAC	1533
ADPGPP	TTATTTTGGGAAT AAT ATAAACTTTGGAGGTGGTTTCGTAGAGGTA
Arabidopsis thaliana	CAAACACTATGACAATAATAACTCTCAGC
Gly163Asn	GCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC	1534
GGC-AAT	CTCCAAAGTTTAT ATT ATTCCCAAAATAAGTTCGTGCTAAATGTCG
	ATTAAGGGAAGCTGAGTTGAACTGTGTCA
	TGGGAAT AAT ATAAACT	1535
	AGTTTAT ATT ATTCCCA	1536

Increased Starch	TGACACAGTTCAACTCAGCTTCCCTTAATCGACATTTAGCACGAAC	1537
ADPGPP	TTATTTTGGGAAT AAC ATAAACTTTGGAGGTGGTTTCGTAGAGGTA
Arabidopsis thaliana	CAAACACTATGACAATAATAACTCTCAGC
Gly163Asn	GCTGAGAGTTATTATTGTCATAGTGTTTGTACCTCTACGAAACCAC	1538
GGC-AAC	CTCCAAAGTTTAT GTT ATTCCCAAAATAAGTTCGTGCTAAATGTCG
	ATTAAGGGAAGCTGAGTTGAACTGTGTCA
	TGGGAAT AAC ATAAACT	1539
	AGTTTAT GTT ATTCCCA	1540

Increased Starch	TTGAGGAACAACCAACGGCAGATCCAAAAGCTGTTGCCTCTGTCA	1541
ADPGPP	TTCTAGGTGGTGGT AAA GGAACTCGTCTTTTTCCTCTTACAAGCA
Lycopersicon	GAAGAGCTAAACCAGCTGTTCCTATTGGTGG
esculentum	CCACCAATAGGAACAGCTGGTTTAGCTCTTCTGCTTGTAAGAGGA	1542
Val94Lys	AAAAGACGAGTTCC TTT ACCACCACCTAGAATGACAGAGGCAACA
GTT-AAA	GCTTTTGGATCTGCCGTTGGTTGTTCCTCAA
	TGGTGGT AAA GGAACTC	1543
	GAGTTCC TTT ACCACCA	1544

Increased Starch	CAAGCAGAAGAGCTAAACCAGCTGTTCCTATTGGTGGTTGTTACC	1545
ADPGPP	GGCTAATTGATGTAC A AATGAGTAACTGCATTAACAGTGGCATAC
Lycopersicon	GGAAAATTTTCATCTTAACACAGTTCAATTC
esculentum	GAATTGAACTGTGTTAAGATGAAAATTTTCCGTATGCCACTGTTAA	1546
Pro122Leu	TGCAGTTACTCATT T GTACATCAATTAGCCGGTAACAACCACCAAT
CCA-CAA	AGGAACAGCTGGTTTAGCTCTTCTGGTTG
	TGATGTAC A AATGAGTA	1547
	TACTCATT T GTACATCA	1548

Increased Starch	CACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA	1549
ADPGPP	TAATTTTGGAAAT AAT GTGGGTTTTGGAGATGGATTTGTGGAGGTT
Lycopersicon	TTAGCTGCAACCCAGACTCCAGGGGATGC
esculentum	GCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA	1550
Gly158Asn	TCTCCAAAACCCAC ATT ATTTCCAAAATTATACGTGCGGGCAAGGT
GGA-AAT	GACGATTGAGGGAAAAGGAATTGAACTGTG
	TGGAAAT AAT GTGGGTT	1551
	AACCCAC ATT ATTTCCA	1552

Increased Starch	CACAGTTCAATTCCTTTTCCCTCAATCGTCACCTTGCCCGCACGTA	1553
ADPGPP	TAATTTTGGAAAT AAC GTGGGTTTTGGAGATGGATTTGTGGAGGT
Lycopersicon	TTTAGCTGCAACCCAGACTCCAGGGGATGC
esculentum	GCATCCCCTGGAGTCTGGGTTGCAGCTAAAACCTCCACAAATCCA	1554
Gly158Asn	TCTCCAAAACCCAC GTT ATTTCCAAAATTATACGTGCGGGCAAGGT
GGA-AAC	GACGATTGAGGGAAAAGGAATTGAACTGTG
	TGGAAAT AAC GTGGGTT	1555
	AACCCAC GTT ATTTCCA	1556

Increased Starch	ACGTAGATTTGGAAAAAAGAGACCCAAGTACAGTTGTAGCAATTAT	1557
ADPGPP	ACTAGGTGGAGGT AAA GGAACTCGTCTCTTCCCTCTCACCAAGCG
Cicer arietinum	ACGAGCCAAGCCTGCTGTTCCAATTGGAGG
Ala101Lys	CCTCCAATTGGAACAGCAGGCTTGGCTCGTCGCTTGGTGAGAGG	1558
GCT-AAA	GAAGAGACGAGTTCC TTT ACCTCCACCTAGTATAATTGCTACAACT
	GTACTTGGGTCTCTTTTTTCCAAATCTACGT
	TGGAGGT AAA GGAACTC	1559
	GAGTTCC TTT ACCTCCA	1560

Increased Starch	CCAAGCGACGAGCCAAGCCTGCTGTTCCAATTGGAGGTGCTTATA	1561
ADPGPP	GGCTGATAGATGTAC T AATGAGTAACTGCATCAATAGTGGGATCA
Cicer arietinum	ACAAAGTATACATTCTCACTCAATTTAATTC
Pro129Leu	GAATTAAATTGAGTGAGAATGTATACTTTGTTGATCCCACTATTGA 1562
CCA-CTA	TGCAGTTACTCATT A GTACATCTATCAGCCTATAAGCACCTCCAAT
	TGGAACAGCAGGCTTGGCTCGTCGCTTGG
	AGATGTAC T AATGAGTA	1563
	TACTCATT A GTACATCT	1564

Increased Starch	CTCAATTTAATTCAGCCTCACTCAACAGGCATATTGCACGTGCTTA	1565
ADPGPP	TAACTCTGGTACT AAT GTCACTTTTGGAGATGGCTATGTTGAGGTT
Cicer arietinum	CTTGCAGCAACTCAAACTCCAGGGGAGCA
Gly165Asn	TGCTCCCGTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA	1566
GGA-AAT	TCTCCAAAAGTGAC ATT AGTACCAGAGTTATAAGCACGTGCAATAT
	GCCTGTTGAGTGAGGCTGAATTAAATTGAG
	TGGTACT AAT GTCACTT	1567
	AAGTGAC ATT AGTACCA	1568

Increased Starch	CTCAATTTAATTCAGCCTCACTCAACAGGCATATTGCACGTGCTTA	1569
ADPGPP	TAACTCTGGTACT AAC GTCACTTTTGGAGATGGCTATGTTGAGGTT
Cicer arietinum	CTTGCAGCAACTCAAACTCCAGGGGAGCA
Gly165Asn	TGCTCCCCTGGAGTTTGAGTTGCTGCAAGAACCTCAACATAGCCA	1570
GGA-AAC	TCTCCAAAAGTGAC GTT AGTACCAGAGTTATAAGCACGTGCAATAT
	GCCTGTTGAGTGAGGCTGAATTAAATTGAG
	TGGTACT AAC GTCACTT	1571
	AAGTGAC GTT AGTACCA	1572

Increased Starch	ATATTGGAGAGGCGTCGGGCAAACCCTAAGAATGTGGCTGCAATC 1573
ADPGPP	ATACTGCCAGGCGGT AA AGGGACACACCTATTCCCTCTCACCAAT
Ipomoea batatas	CGAGCTGCAACCCCTGCTGTTCCACTTGGAG
Ala94Lys	CTCCAAGTGGAACAGCAGGGGTTGCAGCTCGATTGGTGAGAGGG	1574
GCA-AAA	AATAGGTGTGTCCCT TT ACCGCCTGGCAGTATGATTGCAGCCACA
	TTCTTAGGGTTTGCCCGACGCCTCTCCAATAT
	CAGGCGGT AA AGGGACA	1575
	TGTCCCT TT ACCGCCTG	1576

Increased Starch	CCAATCGAGCTGCAACCCCTGCTGTTCCACTTGGAGGATGCTATA	1577
ADPGPP	GGTTGATCGACATTC T AATGAGCAACTGCATCAACAGCGGGGTTA
Ipomoea batatas	ACAAGATCTTTGTGCTGACCCAGTTCAATTC
Pro122Leu	GAATTGAACTGGGTCAGCACAAAGATCTTGTTAACCCCGCTGTTG	1578
CCA-CTA	ATGCAGTTGCTCATT A GAATGTCGATCAACCTATAGCATCCTCCAA
	GTGGAACAGCAGGGGTTGCAGCTCGATTGG
	CGACATTC T AATGAGCA	1579
	TGCTCATT A GAATGTCG	1580

Increased Starch	TGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC	1581
ADPGPP	CGTCTTTGGCAAT AAT GTGAGCTTCGGAGATGGATTTGTTGAGGT
Ipomoea batatas	GCTGGCTGCAACCCAAACACAAGGGGAAAC
Gly157Asn	GTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA	1582
GGT-AAT	TCTCCGAAGCTCAC ATT ATTGCCAAAGACGGTACGGGAAATGTGA
	CGGTTAAGAGAAGCTGAATTGAACTGGGTCA
	TGGCAAT AAT GTGAGCT	1583
	AGCTCAC ATT ATTGCCA	1584

Increased Starch	TGACCCAGTTCAATTCAGCTTCTCTTAACCGTCACATTTCCCGTAC	1585
ADPGPP	CGTCTTTGGCAAT AAC GTGAGCTTCGGAGATGGATTTGTTGAGGT
Ipomoea batatas	GCTGGCTGCAACCCAAACACAAGGGGAAAC
Gly157Asn	GTTTCCCCTTGTGTTTGGGTTGCAGCCAGCACCTCAACAAATCCA	1586
GGT-AAC	TCTCCGAAGCTCAC GTT ATTGCCAAAGACGGTACGGGAAATGTGA
	CGGTTAAGAGAAGCTGAATTGAACTGGGTCA
	TGGCAAT AAC GTGAGCT	1587
	AGCTCAC GTT ATTGCCA	1588

Increased Starch	CATTCCGGAGGAACTTTGCGGATCCAAATGAGGTTGCTGCTGTTA	1589
ADPGPP	TATTGGGTGGTGGCA AA GGGACTCAACTTTTTCCTCTCACAAGCA
Oryza sativa	CAAGGGCCACGCCTGCTGTTCCTATTGGAGG
Thr96Lys	CCTCCAATAGGAACAGCAGGCGTGGCCCTTGTGCTTGTGAGAGG	1590
ACC-AAA	AAAAAGTTGAGTCCC TT TGCCACCACCCAATATAACAGCAGCAAC
	CTCATTTGGATCCGCAAAGTTCCTCCGGAATG
	TGGTGGCA AA GGGACTC	1591
	GAGTCCC TT TGCCACCA	1592

Increased Starch	CAAGCACAAGGGCCACGCCTGCTGTTCCTATTGGAGGATGCTATA	1593
ADPGPP	GGCTTATCGATATCC T CATGAGCAACTGTTTCAACAGTGGCATAAA
Oryza sativa	CAAGATATTCATAATGACTCAATTCAACTC
Pro124Leu	GAGTTGAATTGAGTCATTATGAATATCTTGTTTATGCCACTGTTGA	1594
CCC-CTC	AACAGTTGCTCATG A GGATATCGATAAGCCTATAGCATCCTCCAAT
	AGGAACAGCAGGCGTGGCCCTTGTGCTTG
	CGATATCC T CATGAGCA	1595
	TGCTCATG A GGATATCG	1596

Increased Starch	TGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC	1597
ADPGPP	GTACCTTGGTGGT AAT ATCAACTTTACTGATGGTTCTGTTGAGGTA
Oryza sativa	TTAGCCGCTACACAAATGCCTGGGGAGGC
Gly159Asn	GCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA	1598
GGA-AAT	TCAGTAAAGTTGAT ATT ACCACCAAGGTACGTACGATGAATGTGA
	CGATTAAGAGATGCTGAGTTGAATTGAGTCA
	TGGTGGT AAT ATCAACT	1599
	AGTTGAT ATT ACCACCA	1600

Increased Starch	TGACTCAATTCAACTCAGCATCTCTTAATCGTCACATTCATCGTAC	1601
ADPGPP	GTACCTTGGTGGT AAC ATCAACTTTACTGATGGTTCTGTTGAGGTA
Oryza sativa	TTAGCCGCTACACAAATGCCTGGGGAGGC
Gly159Asn	GCCTCCCCAGGCATTTGTGTAGCGGCTAATACCTCAACAGAACCA	1602
GGA-AAC	TCAGTAAAGTTGAT GTT ACCACCAAGGTACGTACGATGAATGTGA
	CGATTAAGAGATGCTGAGTTGAATTGAGTCA
	TGGTGGT AAC ATCAACT	1603
	AGTTGAT GTT ACCACCA	1604

Increased Starch	GTCCTTCAGGAGGATTAAGCGATCCGAACGAGGTTGCGGCCGTC	1605
ADPGPP	ATACTCGGCGGCGGCA AA GGGACTCAGCTCTTCCCACTCACGAG
Triticum aestivum	CACAAGGGCCACACCTGCTGTTCCTATTGGAGG
Thr80Lys	CCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTCGTGAGTGG	1606
ACC-AAA	GAAGAGCTGAGTCCC TT TGCCGCCGCCGAGTATGACGGCCGCAA
	CCTCGTTCGGATCGCTTAATCCTCCTGAAGGAC
	CGGCGGCA AA GGGACTC	1607
	GAGTCCC TT TGCCGCCG	1608

Increased Starch	CGAGCACAAGGGCCACACCTGCTGTTCCTATTGGAGGATGTTACA	1609
ADPGPP	GGCTCATCGACATTC T CATGAGCAACTGCTTCAACAGTGGCATCA
Triticum aestivum	ACAAGATATTCGTCATGACCCAGTTCAACTC
Pro108Leu	GAGTTGAACTGGGTCATGACGAATATCTTGTTGATGCCACTGTTG	1610
CCC-CTC	AAGCAGTTGCTCATG A GAATGTCGATGAGCCTGTAACATCCTCCA
	ATAGGAACAGCAGGTGTGGCCCTTGTGCTCG
	CGACATTC T CATGAGCA	1611
	TGCTCATG A GAATGTCG	1612

Increased Starch	TGACCCAGTTCAACTCGGCCTCCCTTAATCGTCACATTCACCGCA	1613
ADPGPP	CCTACCTCGGCGGG AAT ATCAATTTCACTGATGGATCCGTTGAGG
Triticum aestivum	TATTGGCCGCGACGCAAATGCCCGGGGAGGC
Gly143Asn	GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC	1614
GGA-AAT	ATCAGTGAAATTGAT ATT CCCGCCGAGGTAGGTGCGGTGAATGTG
	ACGATTAAGGGAGGCCGAGTTGAACTGGGTCA
	CGGCGGG AAT ATCAATT	1615
	AATTGAT ATT CCCGCCG	1616

Increased Starch	TGACCCAGTTCAACTCGGCCTCCCTTAATCGTCACATTCACCGCA	1617
ADPGPP	CCTACCTCGGCGGG AAC ATCAATTTCACTGATGGATCCGTTGAGG
Triticum aestivum	TATTGGCCGCGACGCAAATGCCCGGGGAGGC
Gly143Asn	GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACGGATCC	1618
GGA-AAC	ATCAGTGAAATTGAT GTT CCCGCCGAGGTAGGTGCGGTGAATGTG
	ACGATTAAGGGAGGCCGAGTTGAACTGGGTCA
	CGGCGGG AAC ATCAATT	1619
	AATTGAT GTT CCCGCCG	1620

Increased Starch	CCTCCCGAAAGAATTATGCTGATGCAAGCCACGTTTCTGCTGTCA	1621
ADPGPP	TTTTGGGTGGAGGCA AA GGAGTTCAACTCTTTCCTCTGACAAGCA
Oryza sativa	CAAGGGCTACCCCCGCTGTTCCTGTTGGAGG
Thr95Lys	CCTCCAACAGGAACAGCGGGGGTAGCCCTTGTGCTTGTCAGAGG	1622
ACT-AAA	AAAGAGTTGAACTCC TT TGCCTCCACCCAAAATGACAGCAGAAAC
	GTGGCTTGCATCAGCATAATTCTTTCGGGAGG
	TGGAGGCA AA GGAGTTC	1623
	GAACTCC TT TGCCTCCA	1624

Increased Starch	CAAGCACAAGGGCTACCCCCGCTGTTCCTGTTGGAGGATGTTACA	1625
ADPGPP	GGCTTATTGACATCC T TATGAGCAATTGCTTCAATAGCGGAATAAA
Oryza sativa	TAAAATATTTGTGATGACTCAGTTCAATTC
Pro123Leu	GAATTGAACTGAGTCATCACAAATATTTTATTTATTCCGCTATTGAA	1626
CCT-CTT	GCAATTGCTCATA A GGATGTCAATAAGCCTGTAACATCCTCCAACA
	GGAACAGCGGGGGTAGCCCTTGTGCTTG
	TGACATCC T TATGAGCA	1627
	TGCTCATA A GGATGTCA	1628

Increased Starch	TGACTCAGTTCAATTCTGCTTCTCTTAATCGCCATATCCATCATACA	1629
ADPGPP	TACCTTGGTGGG AAT ATCAACTTTACTGATGGGTCTGTGCAGGTA
Oryza sativa	TTGGCTGCTACACAAATGCCTGACGAACC
Gly158Asn	GGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA	1630
GGG-AAT	TCAGTAAAGTTGAT ATT CCCACCAAGGTATGTATGATGGATATGGC
	GATTAAGAGAAGCAGAATTGAACTGAGTCA
	TGGTGGG AAT ATCAACT	1631
	AGTTGATATTCCCACCA	1632

Increased Starch	TGACTCAGTTCAATTCTGCTTCTCTTAATCGCCATATCCATCATACA	1633
ADPGPP	TACCTTGGTGGG AAC ATCAACTTTACTGATGGGTCTGTGCAGGTA
Oryza sativa	TTGGCTGCTACACAAATGCCTGACGAACC
Gly158Asn	GGTTCGTCAGGCATTTGTGTAGCAGCCAATACCTGCACAGACCCA	1634
GGG-AAC	TCAGTAAAGTTGAT GTT CCCACCAAGGTATGTATGATGGATATGG
	CGATTAAGAGAAGCAGAATTGAACTGAGTCA
	TGGTGGG AAC ATCAACT	1635
	AGTTGAT GTT CCCACCA	1636

Increased Starch	CCTTCCGCAGGAATTACGCCGATCCGAACGAGGTCGCGGCCGTC	1637
ADPGPP	ATACTCGGCGGTGGCAAAGGGACTCAGCTCTTCCCTCTCACAAG
Triticum pestivum	CACAAGGGCCACACCTGCTGTTCCTATTGGAGG
Thr99Lys	CCTCCAATAGGAACAGCAGGTGTGGCCCTTGTGCTTGTGAGAGG	1638
ACC-AAA	GAAGAGCTGAGTCCC TT TGCCACCGCCGAGTATGACGGCCGCGA
	CCTCGTTCGGATCGGCGTAATTCCTGCGGAAGG
	CGGTGGCA AA GGGACTC	1639
	GAGTCCC TT TGCCACCG	1640

Increased Starch	CAAGCACAAGGGCCACACCTGCTGTTCCTATTGGAGGATGTTACA	1641
ADPGPP	GGCTCATCGATATTC T CATGAGCAACTGCTTCAATAGTGGCATCAA
Triticum aestivum	CAAGATATTCGTCATGACGCAGTTCAACTC
Pro127Leu	GAGTTGAACTGCGTCATGACGAATATCTTGTTGATGCCACTATTGA	1642
CCC-CTC	AGCAGTTGCTCATG A GAATATCGATGAGCCTGTAACATCCTCCAA
	TAGGAACAGCAGGTGTGGCCCTTGTGCTTG
	CGATATTC T CATGAGCA	1643
	TGCTCATG A GAATATCG	1644

Increased Starch	TGACGCAGTTCAACTCGGCCTCTCTTAATCGTCACATTCACCGCA	1645
ADPGPP	CCTACCTCGGCGGG AAT ATCAATTTCACTGATGGATCTGTTGAGG
Triticum aestivum	TATTGGCCGCGACGCAAATGCCCGGGGAGGC
Gly162Asn	GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC	1646
GGA-AAT	ATCAGTGAAATTGAT ATT CCCGCCGAGGTAGGTGCGGTGAATGTG
	ACGATTAAGAGAGGCCGAGTTGAACTGCGTCA
	CGGCGGG AAT ATCAATT	1647
	AATTGAT ATT CCCGCCG	1648

Increased Starch	TGACGCAGTTCAACTCGGCCTCTCTTAATCGTCACATTCACCGCA	1649
ADPGPP	CCTACCTCGGCGGG AAC ATCAATTTCACTGATGGATCTGTTGAGG
Triticum aestivum	TATTGGCCGCGACGCAAATGCCCGGGGAGGC
Gly162Asn	GCCTCCCCGGGCATTTGCGTCGCGGCCAATACCTCAACAGATCC	1650
GGA-AAC	ATCAGTGAAATTGAT GTT CCCGCCGAGGTAGGTGCGGTGAATGTG
	ACGATTAAGAGAGGCCGAGTTGAACTGCGTCA
	CGGCGGG AAC ATCAATT	1651
	AATTGAT GTT CCCGCCG	1652

Increased Starch	CTTTTCGGAGGAATTATGCTGATCCTAATGAAGTCGCTGCCGTCA	1653
ADPGPP	TTTTGGGTGGTGGTA AA GGGACTCAGCTTTTCCCTCTCACAAGCA
Zea mays	CAAGGGCCACCCCTGCTGTTCCTATTGGAGG
Thr96Lys	CCTCCAATAGGAACAGCAGGGGTGGCCCTTGTGCTTGTGAGAGG	1654
ACC-AAA	GAAAAGCTGAGTCCC TT TACCACCACCCAAAATGACGGCAGCGAG
	TTCATTAGGATCAGCATAATTCCTCCGAAAAG
	TGGTGGTA AA GGGACTC	1655
	GAGTCCC TT TACCACCA	1656

Increased Starch	CAAGCACAAGGGCCACCCCTGCTGTTCCTATTGGAGGATGTTACA	1657
ADPGPP	GGCTTATTGATATCC T CATGAGCAACTGTTTCAACAGTGGCATAAA
Zea mays	CAAGATATTTGTTATGACTCAGTTCAACTC
Pro124Leu	GAGTTGAACTGAGTCATAACAAATATCTTGTTTATGCCACTGTTGA	1658
CCC-CTC	AACAGTTGCTCATG A GGATATCAATAAGCCTGTAACATCCTCCAAT
	AGGAACAGCAGGGGTGGCCCTTGTGCTTG
	TGATATCC T CATGAGCA	1659
	TGCTCATG A GGATATCA	1660

Increased Starch	TGACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATTCATCGTAC	1661
ADPGPP	CTATCTTGGTGGG AAT ATCAACTTCACTGATGGATCTGTTGAGGT
Zea mays	GCTGGCTGCAACACAAATGCCTGGGGAGGC
Gly159Asn	GCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA	1662
GGG-AAT	TCAGTGAAGTTGAT ATT CCCACCAAGATAGGTACGATGAATGTGA
	CGGTTAAGAGAAGCTGAGTTGAACTGAGTCA
	TGGTGGG AAT ATCAACT	1663
	AGTTGAT ATT CCCACCA	1664

Increased Starch	TGACTCAGTTCAACTCAGCTTCTCTTAACCGTCACATTCATCGTAC	1665
ADPGPP	CTATCTTGGTGGG AAC ATCAACTTCACTGATGGATCTGTTGAGGT
Zea mays	GCTGGCTGCAACACAAATGCCTGGGGAGGC
Gly159Asn	GCCTCCCCAGGCATTTGTGTTGCAGCCAGCACCTCAACAGATCCA	1666
GGG-AAC	TCAGTGAAGTTGAT GTT CCCACCAAGATAGGTACGATGAATGTGA
	CGGTTAAGAGAAGCTGAGTTGAACTGAGTCA
	TGGTGGG AAC ATCAACT	1667
	AGTTGAT GTT CCCACCA	1668

Increased Starch	CTTGAGAGGCAAAAGAAGGGCGATGCAAGGACAGTAGTAGCAAT	1669
ADPGPP	CATTCTAGGAGGGGGA AA GGGAACTCGTCTTTTCCCCCTCACCAA
Solanum tuberosum	ACGTCGTGCTAAGCCTGCCGTTCCAATGGGAG
Ala58Lys	CTCCCATTGGAACGGCAGGCTTAGCACGACGTTTGGTGAGGGGG	1670
GCG-AAG	AAAAGACGAGTTCCC TT TCCCCCTCCTAGAATGATTGCTACTACTG
	TCCTTGCATCGCCCTTCTTTTGCCTCTCAAG
	GAGGGGGA AA GGGAACT	1671
	AGTTCCC TT TCCCCCTC	1672

Increased Starch	CCAAACGTCGTGCTAAGCCTGCCGTTCCAATGGGAGGAGCATATA	1673
ADPGPP	GGCTAATTGATGTAC T AATGAGCAACTGTATTAACAGTGGCATCAA
Solanum tuberosum	CAAAGTATACATTCTCACTCAATTCAACTC
Pro86Leu	GAGTTGAATTGAGTGAGAATGTATACTTTGTTGATGCCACTGTTAA	1674
CCA-CTA	TACAGTTGCTCATT A GTACATCAATTAGCCTATATGCTCCTCCCAT
	TGGAACGGCAGGCTTAGCACGACGTTTGG
	TGATGTAC T AATGAGCA	1675
	TGCTCATT A GTACATCA	1676

Increased Starch	CTCAATTCAACTCAGCCTCACTTAACAGGCATATAGCTCGTGCTTA	1677
ADPGPP	CAACTTTGGCAAT AAT GTCACATTCGAGAGTGGCTATGTCGAGGT
Solanum tuberosum	CTTAGCAGCAACTCAAACACCAGGTGAATT
Gly122Asn	AATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA	1678
GGG-AAT	CTCTCGAATGTGAC ATT ATTGCCAAAGTTGTAAGCACGAGCTATAT
	GCCTGTTAAGTGAGGCTGAGTTGAATTGAG
	TGGCAAT AAT GTCACAT	1679
	ATGTGAC ATT ATTGCCA	1680

Increased Starch	CTCAATTCAACTCAGCCTCACTTAACAGGCATATAGCTCGTGCTTA	1681
ADPGPP	CAACTTTGGCAAT AAC GTCACATTCGAGAGTGGCTATGTCGAGGT
Solanum tuberosum	CTTAGCAGCAACTCAAACACCAGGTGAATT
Gly122Asn	AATTCACCTGGTGTTTGAGTTGCTGCTAAGACCTCGACATAGCCA	1682
GGG-AAC	CTCTCGAATGTGAC GTT ATTGCCAAAGTTGTAAGCACGAGCTATAT
	GCCTGTTAAGTGAGGCTGAGTTGAATTGAG
	TGGCAAT AAC GTCACAT	1683
	ATGTGACGTTATTGCCA	1684

Increased Starch	TATTTGAATCTCCAAAAGCTGACCCAAAAAATGTGGCTGCAATTGT	1685
ADPGPP	GCTGGGTGGTGGT AAA GGGACTCGCCTCTTTCCTCTTACTAGCAG
Beta vulgaris	GAGAGCTAAGCCAGCAGTGCCAATTGGAGG
Ala98Lys	CCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA	1686
GCT-AAA	AAGAGGCGAGTCCC TTT ACCACCACCCAGCACAATTGCAGCCACA
	TTTTTTGGGTCAGCTTTTGGAGATTCAAATA
	TGGTGGT AAA GGGACTC	1687
	GAGTCCC TTT ACCACCA	1688

Increased Starch	TATTTGAATCTCCAAAAGCTGACCCAAAAAATGTGGCTGCAATTGT	1689
ADPGPP	GCTGGGTGGTGGT AAC GGGACTCGCCTCTTTCCTCTTACTAGCAG
Beta vulgaris	GAGAGCTAAGCCAGCAGTGCCAATTGGAGG
Ala98Lys	CCTCCAATTGGCACTGCTGGCTTAGCTCTCCTGCTAGTAAGAGGA	1690
GCT-AAC	AAGAGGCGAGTCCC GTT ACCACCACCCAGCACAATTGCAGCCAC
	ATTTTTTGGGTCAGCTTTTGGAGATTCAAATA
	TGGTGGT AAC GGGACTC	1691
	GAGTCCC GTT ACCACCA	1692

Increased Starch	CTAGCAGGAGAGCTAAGCCAGCAGTGCCAATTGGAGGGTGTTAC	1693
ADPGPP	AGGCTGATTGATGTGC T TATGAGCAACTGCATCAACAGTGGCATT
Beta vulgaris	AGAAAGATTTTCATTCTTACCCAGTTCAATTC
Pro126Leu	GAATTGAACTGGGTAAGAATGAAAATCTTTCTAATGCCACTGTTGA	1694
CCT-CTT	TGCAGTTGCTCATA A GCACATCAATCAGCCTGTAACACCCTCCAA
	TTGGCACTGCTGGCTTAGCTCTCCTGCTAG
	TGATGTGC T TATGAGCA	1695
	TGCTCATA A GCACATCA	1696

Increased Starch	CCCAGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA	1697
ADPGPP	TAATTTTGGAGAT AAT GTGAATTTTGGGGATGGCTTTGTGGAGGTT
Beta vulgaris	TTTGCTGCTACACAAACACCTGGAGAATC
Gly162Asn	GATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA	1698
GGT-AAT	TCCCCAAAATTCAC ATT ATCTCCAAAATTATAGGTTCGAGCAAGAT
	GACGATTAAGCGAAAACGAATTGAACTGGG
	TGGAGAT AAT GTGAATT	1699
	AATTCAC ATT ATCTCCA	1700

Increased Starch	CCCAGTTCAATTCGTTTTCGCTTAATCGTCATCTTGCTCGAACCTA	1701
ADPGPP	TAATTTTGGAGAT AAC GTGAATTTTGGGGATGGCTTTGTGGAGGT
Beta vulgaris	TTTTGCTGCTACACAAACACCTGGAGAATC
Gly162Asn	GATTCTCCAGGTGTTTGTGTAGCAGCAAAAACCTCCACAAAGCCA	1702
GGT-AAC	TCCCCAAAATTCAC GTT ATCTCCAAAATTATAGGTTCGAGCAAGAT
	GACGATTAAGCGAAAACGAATTGAACTGGG
	TGGAGAT AAC GTGAATT	1703
	AATTCAC GTT ATCTCCA	1704

TABLE 21


Oligonucleotides to produce plants with waxy starch

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Waxy starch	GAATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTT	1705
GBSS	CTTCTAACTTTGTGT G AAGAACTTCACTTTTCAACAATCATGGTGCT
Arabidopsis thaliana	TCTTCATGCTCTGATGTCGCTCAGATTAC
Ser12Term	GTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGTTG	1706
TCA-TGA	AAAAGTGAAGTTCTT C ACACAAAGTTAGAAGAAGCAGTCACAGTTG
	CCATTATGAACTACCCGTTTACCTGGATTC
	CTTTGTGT G AAGAACTT	1707
	AAGTTCTT C ACACAAAG	1708

Waxy starch	ATCCAGGTAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTT	1709
GBSS	CTAACTTTGTGTCA T GAACTTCACTTTTCAACAATCATGGTGCTTCT
Arabidopsis thaliana	TCATGCTCTGATGTCGCTCAGATTACCT
Arg13Term	AGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCATGATTGT	1710
AGA-TGA	TGAAAAGTGAAGTTC A TGACACAAAGTTAGAAGAAGCAGTCACAGT
	TGCCATTATGAACTACCCGTTTACCTGGAT
	TTGTGTCA T GAACTTCA	1711
	TGAAGTTC A TGACACAA	1712

Waxy starch	TAAACGGGTAGTTCATAATGGCAACTGTGACTGCTTCTTCTAACTT	1713
GBSS	TGTGTCAAGAACTTGACTTTTCAACAATCATGGTGCTTCTTCATGCT
Arabidopsis thaliana	CTGATGTCGCTCAGATTACCTTAAAAGG
Ser15Term	CCTTTTAAGGTAATCTGAGCGACATCAGAGCATGAAGAAGCACCAT	1714
TCA-TGA	GATTGTTGAAAAGT C AAGTTCTTGACACAAAGTTAGAAGAAGCAGT
	CACAGTTGCCATTATGAACTACCCGTTTA
	AAGAACTT G ACTTTTCA	1715
	TGAAAAGT C AAGTTCTT	1716

Waxy starch	TGACTGCTTCTTCTAACTTTGTGTCAAGAACTTGACTTTTCAACAAT	1717
GBSS	CATGGTGCTTCTT G ATGCTCTGATGTCGCTCAGATTACCTTAAAAG
Arabidopsis thaliana	GCCAATCCTTGACTCATTGTGGGTTAAG
Ser24Term	CTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATCTGAG	1718
TCA-TGA	CGACATCAGAGCAT C AAGAAGCACCATGATTGTTGAAAAGTGAAG
	TTCTTGACACAAAGTTAGAAGAAGCAGTCA
	TGCTTCTT G ATGCTCTG	1719
	CAGAGCAT C AAGAAGCA	1720

Waxy starch	TGCTTCTTCTAACTTTGTGTCAAGAACTTCACTTTTCAACAATCATG	1721
GBSS	GTGCTTCTTCATG A TCTGATGTCGCTCAGATTACCTTAAAAGGCCA
Arabidopsis thaliana	ATCCTTGACTCATTGTGGGTTAAGGTCA
Cys25Term	TGACCTTAACCCACAATGAGTCAAGGATTGGCCTTTTAAGGTAATC	1722
TGC-TGA	TGAGCGACATCAGATCATGAAGAAGCACCATGATTGTTGAAAAGT
	GAAGTTCTTGACACAAAGTTAGAAGAAGCA
	TCTTCATG A TCTGATGT	1723
	ACATCAGA T CATGAAGA	1724

Waxy starch	GTAACAGCTTCACAGTTGGTGTCACATGTCCATGGTGGAGCAACG	1725
GBSS	TCTTCACCGGATACT T AAACAAACTTGGCCCAGGTTGGCCTCAGG
Antirrhinum majus	AACCAGCAATTCACTCACAATGGGTTGAGAT
Lys24Term	ATCTCAAGCCATTGTGAGTGAATTGCTGGTTCGTGAGGCCAACCTG	1726
AAA-TAA	GGCCAAGTTTGTTT A AGTATCGGGTGAAGACGTTGCTCCACCATG
	GACATGTGACACCAACTGTGAAGGTGTTAC
	CGGATACT T AAACAAAC	1727
	GTTTGTTT A AGTATCCG	1728

Waxy starch	CACAGTTGGTGTCACATGTCCATGGTGGAGCAAGGTCTTCACCGG	1729
GBSS	ATAGTAAAACAAACT A GGGCGAGGTTGGCCTCAGGAACCAGCAAT
Antirrhinum majus	TCACTCACAATGGGTTGAGATCAATAAACAT
Leu27Term	ATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTTCCTGA	1730
TTG-TAG	GGCCAACCTGGGCC T AGTTTGTTTTAGTATCGGGTGAAGACGTTG
	CTCCACCATGGACATGTGACACCAACTGTG
	AACAAACT A GGCCCAGG	1731
	CCTGGGCC T AGTTTGTT	1732

Waxy starch	TTGGTGTCACATGTCCATGGTGGAGCAACGTCTTCACCGGATACT	1733
GBSS	AAAACAAACTTGGCC T AGGTTGGCCTCAGGAACCAGCAATTCACT
Antirrhinum majus	CACAATGGGTTGAGATCAATAAACATGGTTG
Gln29Term	CAACCATGTTTATTGATCTCAACCCATTGTGAGTGAATTGCTGGTT	1734
GAG-TAG	CCTGAGGCCAACCT A GGCCAAGTTTGTTTTAGTATCCGGTGAAGA
	CGTTGCTCCACCATGGACATGTGACACCAA
	ACTTGGCC T AGGTTGGC	1735
	GCCAACCT A GGCCAAGT	1736

Waxy starch	GGTGGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAG	1737
GBSS	GTTGGCCTCAGGAACTAGCAATTCACTCACAATGGGTTGAGATCA
Antirrhinum majus	ATAAACATGGTTGATAAGCTTCAAATGAGGA
Gln35Term	TCCTCATTTGAAGCTTATCAACCATGTTTATTGATGTCAACCCATTG	1738
GAG-TAG	TGAGTGAATTGCT A GTTCCTGAGGCCAACCTGGGCCAAGTTTGTTT
	TAGTATCCGGTGAAGACGTTGCTCCACC
	TCAGGAAC T AGCAATTC	1739
	GAATTGCT A GTTCCTGA	1740

Waxy starch	GGAGCAACGTCTTCACCGGATACTAAAACAAACTTGGCCCAGGTT	1741
GBSS	GGCCTCAGGAACCAG T AATTCACTCACAATGGGTTGAGATCAATAA
Antirrhinum majus	ACATGGTTGATAAGCTTCAAATGAGGAACA
Gln36Term	TGTTCCTCATTTGAAGCTTATCAACCATGTTTATTGATCTCAACCCA	1742
CAA-TAA	TTGTGAGTGAATT A CTGGTTCCTGAGGCCAACCTGGGCCAAGTTT
	GTTTTAGTATCCGGTGAAGACGTTGCTCC
	GGAACCAG T AATTCACT	1743
	AGTGAATT A CTGGTTCC	1744

Waxy starch	GTGATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTG	1745
GBSS	GGGGTGCCACTTCTTGAGAATCAAAAGTGGGGTTGGGTCAATTAG
Ipomoea batatas	CCCTGAGGAGCCAAGCTGTGACTCACAATG
Gly20Term	CATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCAACC	1746
GGA-TGA	CCACTTTTGATTCTC A AGAAGTGGCACCCCCACAGACATGAGAAA
	CAAAGTGTGAGGCAGTTATAGTCGCCATCAC
	CCACTTCT T GAGAATCA	1747
	TGATTCTC A AGAAGTGG	1748

Waxy starch	ATGGCGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGG	1749
GBSS	GTGCCACTTCTGGA T AATCAAAAGTGGGGTTGGGTCAATTAGCCC
Ipomoea batatas	TGAGGAGCCAAGCTGTGACTCACAATGGGT
Glu21Term	ACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGACCCA	1750
GAA-TAA	ACCCCACTTTTGATT A TCCAGAAGTGGCACCCCCACAGACATGAG
	AAACAAAGTGTGAGGCAGTTATAGTCGCCAT
	CTTCTGGA T AATCAAAA	1751
	TTTTGATT A TCCAGAAG	1752

Waxy starch	CGACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGC	1753
GBSS	CACTTCTGGAGAAT G AAAAGTGGGGTTGGGTCAATTAGCCCTGAG
Ipomoea batatas	GAGCCAAGCTGTGACTCACAATGGGTTGAG
Ser22Term	CTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATTGA	1754
TCA-TGA	CCCAACCCCACTTTT C ATTCTCCAGAAGTGGCACCCCCACAGACAT
	GAGAAACAAAGTGTGAGGCAGTTATAGTCG
	TGGAGAAT G AAAAGTGG	1755
	CCACTTTT C ATTCTCCA	1756

Waxy starch	ACTATAACTGCCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCA	1757
GBSS	CTTCTGGAGAATCA T AAGTGGGGTTGGGTCAATTAGCCCTGAGGA
Ipomoea batatas	GCCAAGCTGTGACTCACAATGGGTTGAGAC
Lys23Term	GTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCAGGGCTAATT	1758
AAA-TAA	GACCCAACCCCACTT A TGATTCTCCAGAAGTGGCACCCCCACAGA
	CATGAGAAACAAAGTGTGAGGCAGTTATAGT
	GAGAATCA T AAGTGGGG	1759
	CCCCACTT A TGATTCTC	1760

Waxy starch	CCTCACACTTTGTTTCTCATGTCTGTGGGGGTGCCACTTCTGGAGA	1761
G BSS	ATCAAAAGTGGGGT A GGGTCAATTAGCCCTGAGGAGCCAAGCTGT
Ipomoea batatas	GACTCACAATGGGTTGAGACCTGTGAACAA
Leu26Term	TTGTTCACAGGTCTCAACCCATTGTGAGTCACAGCTTGGCTCCTCA	1762
TTG-TAG	GGGCTAATTGACCC T ACCCCACTTTTGATTCTCCAGAAGTGGCACC
	CCCACAGACATGAGAAACAAAGTGTGAGG
	AGTGGGGT A GGGTCAAT	1763
	ATTGACCC T ACCCCACT	1764

Waxy starch	CATCGGCGATTGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACG	1765
GBSS	GTGACGGGGTCTTA G GTGGTGTCGAGAAGCGCGTGCTTCAATTCC
Astragalus	CAGGGAAGAACAGAAGCCAAAGTGAATTCA
membranaeus	TGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGAAGCACGCG	1766
Tyr8Term	CTTCTCGACACCAC C TAAGACCCCGTCACCGTTGCCATTCTGTGA
TAT-TAG	GAGAGCAGTAAGGAGCAACAATCGCCGATG
	GGGTCTTA G GTGGTGTC	1767
	GACACCAC C TAAGACCC	1768

Waxy starch	ATTGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACGGTGACGG	1769
GBSS	GGTCTTATGTGGTGT A GAGAAGCGCGTGCTTCAATTCCCAGGGAA
Astragalus	GAACAGAAGCCAAAGTGAATTCACCTCAGAA
membranaeus	TTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATTGA	1770
Ser11Term	AGCACGCGCTTCTCTACACCACATAAGACCCCGTCACCGTTGCCA
TCG-TAG	TTCTGTGAGAGAGCAGTAAGGAGCAACAAT
	TGTGGTGT A GAGAAGCG	1771
	CGCTTCTC T ACACCACA	1772

Waxy starch	TGTTGCTCCTTACTGCTCTCTCACAGAATGGCAACGGTGACGGGG	1773
GBSS	TCTTATGTGGTGTCGTGAAGCGCGTGCTTCAATTCCCAGGGAAGA
Astragalus	ACAGAAGCCAAAGTGAATTCACCTCAGAAGA
membranaeus	TCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTCCCTGGGAATT	1774
Arg12Term	GAAGCACGCGCTTC A CGACACCACATAAGACCCCGTCACCGTTGC
AGA-TGA	CATTCTGTGAGAGAGCAGTCAGGAGCAACA
	TGGTGTCG T GAAGCGCG	1775
	CGCGCTTC A CGACACCA	1776

Waxy starch	ACTGCTCTCTCACAGAATGGCAACGGTGACGGGGTCTTATGTGGT	1777
GBSS	GTCGAGAAGCGCGTG A TTCAATTCCCAGGGAAGAACAGAAGCCAA
Astragalus	AGTGAATTCACCTCAGAAGATAAATCTGAAT
membranaeus	ATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCTTCTGTTCTTC	1778
Cys15Term	CCTGGGAATTGAATCACGCGCTTCTCGACACCACATAAGACCCCG
TGC-TGA	TCACCGTTGCCATTCTGTGAGAGAGCAGT
	AGCGCGTG A TTCAATTC	1779
	GAATTGAA T CACGCGCT	1780

Waxy starch	CACAGAATGGCAACGGTGACGGGGTCTTATGTGGTGTCGAGAAG	1781
GBSS	CGCGTGGTTCAATTCCTAGGGAAGAACAGAAGCCAAAGTGAATTC
Astragalus	ACCTCAGAAGATAAATCTCAATAGCCAAGCAT
membranaeus	ATGCTTGGCTATTGAGATTTATCTTCTGAGGTGAATTCACTTTGGCT	1782
Gln19Term	TCTGTTCTTCCCTAGGAATTGAAGCACGCGCTTCTCGACACCACAT
CAG-TAG	AAGACCCCGTCACCGTTGCCATTCTGTG
	TCAATTCC T AGGGAAGA	1783
	TCTTCCCT A GGAATTGA	1784

Waxy starch	TGTAGCTTGGTAGATTCCCCTTTTTGTCGACCACACATCACATGGC	1785
GBSS	AAGCATCACAGCTT G ACACCACTTTGTGTCAAGAAGCCAAACTTCA
Solanum tuberosum	CTAGACACCAAATCAACCTTGTCACAGAT
Ser7Term	ATCTGTGACAAGGTTGATTTGGTGTCTAGTGAAGTTTGGCTTCTTG	1786
TCA-TGA	ACACAAAGTGGTGT C AAGCTGTGATGCTTGCCATGTGATGTGTGG
	TCTACAAAAAGGGGAATCTACCAAGCTACA
	CACAGCTT G ACACCACT	1787
	AGTGGTGT C AAGCTGTG	1788

Waxy starch	TCCCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTC	1789
GBSS	ACACCACTTTGTGT G AAGAAGCCAAACTTCACTAGACACCAAATCA
Solanum tuberosum	ACCTTGTCACAGATAGGACTCAGGAACCA
Ser12Term	TGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAGTG	1790
TCA-TGA	AAGTTTGGCTTCTT C ACACAAAGTGGTGTGAAGCTGTGATGCTTGC
	CATGTGATGTGTGGTCTACAAAAAGGGGA
	CTTTGTGT G AAGAAGCC	1791
	GGCTTCTT C ACACAAAG	1792

Waxy starch	CCCTTTTTGTAGACCACACATCACATGGCAAGCATCACAGCTTCAC	1793
GBSS	ACCACTTTGTGTCATGAAGCCAAACTTCACTAGACACCAAATCAAC
Solanum tuberosum	CTTGTCACAGATAGGACTCAGGAACCATA
Arg13Term	TATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGTGTCTAG	1794
AGA-TGA	TGAAGTTTGGCTTC A TGACACAAAGTGGTGTGAAGCTGTGATGCTT
	GCCATGTGATGTGTGGTCTACAAAAAGGG
	TTGTGTCA T GAAGCCAA	1795
	TTGGCTTC A TGACACAA	1796

Waxy starch	TTGTAGACCACACATCACATGGCAAGCATCACAGCTTCACACCACT	1797
GBSS	TTGTGTCAAGAAGCTAAACTTCACTAGACACCAAATCAACCTTGTC
Solanum tuberosum	ACAGATAGGACTCAGGAACCATACTCTGA
Gln15Term	TCAGAGTATGGTTCCTGAGTCCTATCTGTGACAAGGTTGATTTGGT	1798
CAA-TAA	GTCTAGTGAAGTTT A GCTTCTTGACACAAAGTGGTGTGAAGCTGTG
	ATGCTTGCCATGTGATGTGTGGTCTACAA
	CAAGAAGC T AAACTTCA	1799
	TGAAGTTT A GCTTCTTG	1800

Waxy starch	CCACACATCACATGGCAAGCATCACAGCTTCACACCACTTTGTGTC	1801
GBSS	AAGAAGCCAAACTT G ACTAGACACCAAATCAACCTTGTCACAGATA
Solanum tuberosum	GGACTCAGGAACCATACTCTGACTCACAA
Sen17Term	TTGTGAGTCAGAGTCTGGTTCCTGAGTCCTATCTGTGACAAGGTTG	1802
TCA-TGA	ATTTGGTGTCTAGT C AAGTTTGGCTTGTTGACACAAAGTGGTGTGA
	AGCTGTGATGCTTGCCATGTGATGTGTGG
	CCAAACTT G ACTAGACA	1803
	TGTCTAGT C AAGTTTGG	1804

Waxy starch	GTCGATCACTCTTCTCTCACCGCCGAAACAGATTTTGACACAAAAA	1805
GBSS	TGGCAACAATAACG T GATCTTCAATGCCGACGAGAACCGCGTGCT
Pisum sativum	TCAATTACCAAGGAAGATCAGCAGAGTCTA
Gly6Term	TAGACTCTGCTGATCTTCCTTGGTCATTGAAGCACGCGGTTCTCGT	1806
GGA-TGA	CGGCATTGAAGATC A CGTTATTGTTGCCATTTTTGTGTCAAAATCT
	GTTTCGGCGGTGAGAGAAGAGTGATCGAC
	CAATAACG T GATCTTCA	1807
	TGAAGATC A CGTTATTG	1808

Waxy starch	ACTCTTCTCTCACCGCCGAAACAGATTTTGACACAAAAATGGCAAC	1809
GBSS	AATAACGGGATCTT G AATGCCGACGAGAACCGCGTGCTTCAATTA
Pisum sativum	CCAAGGAAGATCAGCAGAGTCTAAACTGAA
Ser8Term	TTCAGTTTAGACTCTGCTGATCTTCCTTGGTCATTGAAGCACGCGG	1810
TCA-TGA	TTCTCGTCGGCATT C AAGATCCCGTTATTGTTGCCATTTTTGTGTCA
	AAATCTGTTTCGGCGGTGAGAGAAGAGT
	GGGATCTT G AATGCCGA	1811
	TCGGCATT C AAGATCCC	1812

Waxy starch	ACCGCCGAAACAGATTTTGACACAAAAATGGCAACAATAACGGGA	1813
GBSS	TCTTCAATGCCGACG T GAACCGCGTGCTTCAATTACCAAGGAAGA
Pisum sativum	TCAGCAGAGTCTAAACTGAATTTGCCTCAGA
Arg12Term	TCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTTCCTTGGTCATT	1814
AGA-TGA	GAAGCACGCGGTTC A CGTCGGCATTGAAGATCCCGTTATTGTTGC
	CATTTTTGTGTCAAAATCTGTTTCGGCGGT
	TGCCGACG T GAACCGCG	1815
	CGCGGTTC A CGTCGGCA	1816

Waxy starch	AGATTTTGACACAAAAATGGCAACAATAACGGGATCTTCAATGCCG	1817
GBSS	ACGAGAACCGCGTG A TTCAATTACCAAGGAAGATCAGCAGAGTCT
Pisum sativum	AAACTGAATTTGCCTCAGATACACTTCAAT
Cys15Term	ATTGAAGTGTCTCTGAGGCAAATTCAGTTTAGACTCTGCTGATCTT	1818
TGC-TGA	CCTTGGTCATTGAA T CACGCGGTTCTCGTCGGCATTGAAGATCCC
	GTTATTGTTGCCATTTTTGTGTCAAAATCT
	ACCGCGTG A TTCAATTA	1819
	TAATTGAA T CACGCGGT	1820

Waxy starch	CACAAAAATGGCAACAATAACGGGATCTTCAATGCCGACGAGAAC	1821
GBSS	CGCGTGCTTCAATTA G CAAGGAAGATCAGCAGAGTCTAAACTGAA
Pisum sativum	TTTGCCTCAGATACACTTCAATAACAACCAA
Tyr18Term	TTGGTTGTTATTGAAGTGTATCTGAGGCAAATTCAGTTTAGACTCT	1822
TAC-TAG	GCTGATCTTCCTTG C TAATTGAAGCACGCGGTTCTCGTCGGCATTG
	AAGATCCCGTTATTGTTGCCATTTTTGTG
	TTCAATTA G CAAGGAAG	1823
	CTTCCTTG C TAATTGAA	1824

Waxy starch	TCTACACCGGAGAGAGCACCATGGCAACTGTAATAGCTGCACATT	1825
GBSS	TCGTTTCCAGGAGCT G ACACTTGAGCATCCATGCATTAGAGACTAA
Manihot esculenta	GGCTAATAATTTGTCTCACACTGGACCCTG
Ser14Term	CAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTAATGCAT	1826
TCA-TGA	GGATGCTCAAGTGT C AGCTCCTGGAAACGAAATGTGCAGCTATTA
	CAGTTGCCATGGTGCTCTCTCCGGTGTAGA
	CAGGAGCT G ACACTTGA	1827
	TCAAGTGT C AGCTCCTG	1828

Waxy starch	CCGGAGAGAGCACCATGGCAACTGTAATAGCTGCACATTTCGTTT	1829
GBSS	CCAGGAGCTCACACT A GAGCATCCATGCATTAGAGACTAAGGCTA
Manihot esculenta	ATAATTTGTCTCACACTGGACCCTGGACCCA
Leu16Term	TGGGTCCAGGGTCCAGTGTGAGACAAATTATTAGCCTTAGTCTCTA	1830
TTG-TAG	ATGCATGGATGCTC T AGTGTGAGCTCCTGGAAACGAAATGTGCAG
	CTATTACAGTTGCCATGGTGCTCTCTCCGG
	CTCACACT A GAGCATCC	1831
	GGATGCTC T AGTGTGAG	1832

Waxy starch	TGGCAACTGTAATAGCTGCACATTTCGTTTCCAGGAGCTCACACTT	1833
GBSS	GAGCATCCATGCAT G AGAGACTAAGGCTAATAATTTGTCTCACACT
Manihot esculenta	GGACCCTGGACCCAAACTATCACTCCCAA
Leu21Term	TTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAATTA	1834
TTA-TGA	TTAGCCTTAGTCTCT C ATGCATGGATGCTCAAGTGTGAGCTCCTGG
	AAACGAAATGTGCAGCTATTACAGTTGCCA
	CCATGCAT G AGAGACTA	1835
	TAGTCTCT C ATGCATGG	1836

Waxy starch	GCAACTGTAATAGCTGCACATTTCGTTTCCAGGAGCTCACACTTGA	1837
GBSS	GCATCCATGCATTA T AGACTAAGGCTAATAATTTGTCTCACACTGG
Manihot esculenta	ACCCTGGACCCAAACTATCACTCCCAATG
Glu22Term	CATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAGACAAAT	1838
GAG-TAG	TATTAGCCTTAGTCT A TAATGCATGGATGCTCAAGTGTGAGCTCCT
	GGAAACGAAATGTGCAGCTATTACAGTTGC
	ATGCATTA T AGACTAAG	1839
	CTTAGTCT A TAATGCAT	1840

Waxy starch	GTCATAGCTGCACATTTCGTTTCCAGGAGCTCACACTTGAGCATCC	1841
GBSS	ATGCATTAGAGACT T AGGCTAATAATTTGTCTCACACTGGACCCTG
Manihot esculenta	GACCCAAACTATCACTCCCAATGGTTTAA
Lys24Term	TTAAACCATTGGGAGTGATAGTTTGGGTCCAGGGTCCAGTGTGAG	1842
AAG-TAG	ACAAATTATTAGCCT A AGTCTCTAATGCATGGATGCTCAAGTGTGA
	GCTCCTGGAAACGAAATGTGCAGCTATTAC
	TAGAGACT T AGGCTAAT	1843
	ATTAGCCT A AGTCTCTA	1844

Waxy starch	ACAACTCCTCCGTCACCGGTATAAGCATGGCAACGGTATCGATGG	1845
GBSS	CATCGTGCGTGGCGT G AAAAGGCGCGTGGAGTACAGAGACAAAA
Phaseolus vulgaris	GTGAAATCTTCGGGTCAGATGAGCCTGAACCG
Ser12Term	CGGTTCAGGCTCATCTGACCCGAAGATTTCACTTTTGTCTCTGTCC	1846
TCA-TGA	TCCACGCGCCTTTT C ACGCCACGCACGATGCCATCGATACCGTTG
	CCATGCTTATACCGGTGACGGAGGAGTTGT
	CGTGGCGT G AAAAGGCG	1847
	CGCCTTTT C ACGCCACG	1848

Waxy starch	CACCGGTCTAAGCATGGCAACGGTATCGATGGCATCGTGCGTGGC	1849
GBSS	GTCAAAAGGCGCGTG A AGTACAGAGACAAAAGTGAAATCTTCGGG
Phaseolus vulgaris	TCAGATGAGCCTGAACCGTCATGAATTGAAA
Trp16Term	TTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGATTTCACT	1850
TGG-TGA	TTTGTCTCTGTACT T CACGCGCCTTTTGACGCCACGCACGATGCCA
	TCGATACCGTTGGCATGCTTATACCGGTG
	GGCGCGTG A AGTACAGA	1851
	TCTGTACT T CACGCGCC	1852

Waxy starch	ATAAGCATGGCAACGGTCTCGATGGCATCGTGCGTGGCGTCAAAA	1853
GBSS	GGCGCGTGGAGTACA T AGACAAAAGTGAAATCTTCGGGTCAGATG
Phaseolus vulgaris	AGCCTGAACCGTCATGAATTGAAATACGATG
Glu19Term	CATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACCCGAAGA	1854
GAG-TAG	TTTCACTTTTGTCT A TGTACTCCACGCGCCTTTTGACGCCACGCAC
	GATGCCATCGATACCGTTGCCATGCTTAT
	GGAGTACA T AGACAAAA	1855
	TTTTGTCT A TGTACTCC	1856

Waxy starch	ATGGCAACGGTATCGATGGCATCGTGCGTGGGGTCAAAAGGCGC	1857
GBSS	GTGGAGTACAGAGACA T AAGTGAAATCTTCGGGTCAGATGAGCCT
Phaseolus vulgaris	GAACCGTCATGAATTGAAATACGATGGGTTGA
Lys21Term	TCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCATCTGACC	1858
AAA-TAA	CGAAGATTTCACTT A TGTCTCTGTACTCCACGCGCCTTTTGACGCC
	ACGCACGATGCCATCGATACCGTTGCCAT
	CAGAGACA T AAGTGAAA	1859
	TTTCACTT A TGTCTCTG	1860

Waxy starch	ACGGTATCGATGGCATCGTGCGTGGCGTCAAAAGGCGCGTGGAG	1861
GBSS	TACAGAGACAAAAGTG T AATCTTCGGGTCAGATGAGCCTGAACCG
Phaseolus vulgaris	TCATGAATTGAAATACGATGGGTTGAGATCTC
Lys23Term	GAGATCTCAACCCATCGTATTTCAATTCATGACGGTTCAGGCTCAT	1862
AAA-TAA	CTGACCCGAAGATT A CACTTTTGTCTCTGTACTCCACGCGCCTTTT
	GACGCCACGCACGATGCCATCGATACCGT
	CAAAAGTG T AATCTTCG	1863
	CGAAGATT A CACTTTTG	1864

Waxy starch	GCGCCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGG	1865
GBSS	GTTCCATTCCTAATTA G TGTTCTTATCAAACAAACAGTGTTGGTTCA
Triticum aestivum	CTGAAACTGTCGCCTCACATCCAATTCCAG
Tyr7Term	CTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACTGTT	1866
TAT-TAG	TGTTTGATAAGAACA C TAATTAGGAATGGAACCCATTGGTGCAGCC
	TCTCAATGACGACCTTTTCGAGCTAGGCGC
	CCTAATTA G TGTTCTTA	1867
	TAAGAACA C TAATTAGG	1868

Waxy starch	CCTAGCTCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTC	1869
GBSS	CATTCCTAATTATTG A TCTTATCAAACAAACAGTGTTGGTTCACTGA
Triticum aestivum	AACTGTCGCCTCACATCCAATTCCAGCAA
Cys8Term	TTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACCAACACT	1870
TGT-TGA	GTTTGTTTGATAAGA T CAATAATTAGGAATGGAACCCATTGGTGCA
	GCCTCTCAATGACGACCTTTTCGAGCTAGG
	AATTATTG A TCTTATCA	1871
	TGATAAGA T CAATAATT	1872

Waxy starch	TCGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCC	1873
GBSS	TAATTATTGTTCTTA G CAAACAAACAGTGTTGGTTCACTGAAACTGT
Triticum aestivum	CGCCTCACATCCAATTCCAGCAATCTTGT
Tyr10Term	ACAAGATTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAACC	1874
TAT-TAG	AACACTGTTTGTTTG C TAAGAACAATAATTAGGAATGGAACCCATT
	GGTGCAGCCTCTCAATGACGACCTTTTCGA
	TGTTCTTA G CAAACAAA	1875
	TTTGTTTG C TAAGAACA	1876

Waxy starch	CGAAAAGGTCGTCATTGAGAGGCTGCACCAATGGGTTCCATTCCT	1877
GBSS	AATTATTGTTCTTAT T AAACAAACAGTGTTGGTTCACTGAAACTGTC
Triticum aestivum	GCCTCACATCCAATTCCAGCAATCTTGTA
Gln11Term	TACAAGATTGCTGGAATTGGATGTGAGGCGACAGTTTCAGTGAAC	1878
CAA-TAA	CAACACTGTTTGTTT A ATAAGAACAATAATTAGGAATGGAACCCATT
	GGTGCAGCCTCTCAATGACGACCTTTTCG
	GTTCTTAT T AAACAAAC	1879
	GTTTGTTT A ATAAGAAC	1880

Waxy starch	AGGCTGCACCAATGGGTTCCATTCCTAATTATTGTTCTTATCAAACA	1881
GBSS	AACAGTGTTGGTT G ACTGAAACTGTCGCCTCACATCCAATTCCAGC
Triticum aestivum	AATCTTGTCACAATGAAGTTATGTTCCT
Ser17Term	AGGAACATAACTTCATTGTTACAAGATTGCTGGAATTGGATGTGAG	1882
TCA-TGA	GCGACAGTTTCAGT C AACCAACACTGTTTGTTTGATAAGAACAATA
	ATTAGGAATGGAACCCATTGGTGCAGCCT
	TGTTGGTT G ACTGAAAC	1883
	GTTTCAGT C AACCAACA	1884

Waxy starch	CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT	1885
GBSS	CCGGCGTGCAGGTTTC T AGGGCGTGAGGCCCCGGAGCCCGGCG
Triticum aestivum	GATGCGGCTCTCGGCATGAGGACCGTCGGAGCTA
Gln28Term	TAGCTCCGACGGTCCTCATGCCGAGAGCCGCATCCGCCGGGCTC	1886
CAG-TAG	CGGGGCCTCACGCCCT A GAAACCTGCACGCCGGAACCTGTCGGT
	GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG
	CAGGTTTC T AGGGCGTG	1887
	CACGCCCT A GAAACCTG	1888

Waxy starch	GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCGGATGCGGCTCT	1889
GBSS	CGGCATGAGGACCGTC T GAGCTAGCGCCGCCCCAACGCAAAGCC
Triticum aestivum	GGAAAGCGCACCGCGGGACCCGGCGGTGCCTCT
Gly46Term	AGAGGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGCGTT	1890
GGA-TGA	GGGGCGGCGCTAGCTC A GACGGTCCTCATGCCGAGAGCCGCATC
	CGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC
	GGACCGTC T GAGCTAGC	1891
	GCTAGCTC A GACGGTCC	1892

Waxy starch	CGGAGCCCGGCGGATGCGGCTCTCGGCATGAGGACCGTCGGAG	1893
GBSS	CTAGCGCCGCCCCAACG T AAAGCCGGAAAGCGCACCGCGGGACC
Triticum aestivum	CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACCG
Gln53Term	CGGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG	1894
CAA-TAA	GTGCGCTTTCCGGCTTT A CGTTGGGGCGGCGCTAGCTCCGACGG
	TCCTCATGCCGAGAGCCGCATCCGCCGGGCTCCG
	CCCCAACG T AAAGCCGG	1895
	CCGGCTTT A CGTTGGGG	1896

Waxy starch	GCGGATGCGGCTCTCGGCATGAGGACCGTCGGAGCTAGCGCCGC	1897
GBSS	CCCAACGCAAAGCCGG T AAGCGCACCGCGGGACCCGGCGGTGC
Triticum aestivum	CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCG
Lys56Term	CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAGGCACCGCCG	1898
AAA-TAA	GGTCCCGCGGTGCGCTT A CCGGCTTTGCGTTGGGGCGGCGCTAG
	CTCCGACGGTCCTCATGCCGAGAGCCGCATCCGC
	AAAGCCGG T AAGCGCAC	1899
	GTGCGCTT A CCGGCTTT	1900

Waxy starch	CTCTCCATGGTGGTGCGCGCCACCGGCAGCGGCGGCATGAACCT	1901
GBSS	CGTGTTCGTCGGCGCC T AGATGGCGCCCTGGACCAAGACCGGCG
Triticum aestivum	GCCTCGGCGACGTCCTCGGGGGCCTCCCCCCAG
Glu85Term	CTGGGGGGAGGCCCCCGAGGACGTCGCCGAGGCCGCCGGTCTT	1902
GAG-TAG	GCTCCAGGGCGCCATCT A GGCGCCGACGAACACGAGGTTCATGC
	CGCCGCTGCCGGTGGCGCGCACCACCATGGAGAG
	TCGGCGCC T AGATGGCG	1903
	CGCCATCT A GGCGCCGA	1904

Waxy starch	GTGGTCTCTCGCTGCAGGTAGCCACACCCTGCGCGCGCGATGGC	1905
GBSS	GGCTCTGGTCACGTCG T AGCTCGCCACCTCCGGCACCGTCCTCG
Triticum aestivum	GCATCACCGACAGGTTCCGGCGTGCAGGTTTTC
Gln8Term	GAAAACCTGCACGCCGGAACCTGTCGGTGATGCCGAGGACGGTG	1906
CAG-TAG	CCGGAGGTGGCGAGCT A CGACGTGACCAGAGCCGCCATCGCGC
	GCGCAGGGTGTGGCTACCTGCAGCGAGAGACGAC
	TCACGTCG T AGCTCGCC	1907
	GGCGAGCT A CGACGTGA	1908

Waxy starch	CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT	1909
GBSS	CCGGCGTGCAGGTTTT T AGGGTGTGAGGCCCCGGAGCCCGGCAG
Triticum aestivum	ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA
Gln28Term	TCGCTCCGGTCGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC	1910
GAG-TAG	CGGGGCCTCACACCCT A AAAACCTGCACGCCGGAACCTGTCGGT
	GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG
	CAGGTTTT T AGGGTGTG	1911
	CACACCCT A AAAACCTG	1912

Waxy starch	CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGG	1913
GBSS	AGCGAGCGCCGCCCCG T AGCAACAAAGCCGGAAAGCGCACCGCG
Triticum aestivum	GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCG
Lys52Term	CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTGCGC	1914
AAG-TAG	TTTCCGGCTTTGTTGCT A CGGGGCGGCGCTCGCTCCGGTAGTCCT
	CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG
	CCGCCCCG T AGCAACAA	1915
	TTGTTGCT A CGGGGCGG	1916

Waxy starch	CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG	1917
GBSS	CGAGCGCCGCCCCGAAG T AACAAAGCCGGAAAGCGCACCGCGG
Triticum aestivum	GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA
Gln53Term	TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTG	1918
CAA-TAA	CGCTTTCCGGCTTTGTT A CTTCGGGGCGGCGCTCGCTCCGGTAGT
	CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG
	CCCCGAAG T AACAAAGC	1919
	GCTTTGTT A CTTCGGGG	1920

Waxy starch	AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG	1921
GBSS	CGCCGCCCCGAAGCAA T AAAGCCGGAAAGCGCACCGCGGGACCC
Triticum aestivum	GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG
Gln54Term	CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG	1922
CAA-TAA	GTGCGCTTTCCGGCTTT A TTGCTTCGGGGCGGCGCTCGCTCCGGT
	AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT
	CGAAGCAA T AAAGCCGG	1923
	CCGGCTTT A TTGCTTCG	1924

Waxy starch	CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT	1925
GBSS	CCGGCGTGCAGGTTTC T AGGGCGTGAGGCCCCGGAACCCGGCG
Triticum durum	GATGCGGCCCTCGTCATGAGGACTATCGGAGCGA
Gln28Term	TCGCTCCGATAGTCCTCATGACGAGGGCCGCATCCGCCGGGTTC	1926
CAG-TAG	CGGGGCCTCACGCCCT A GAAACCTGCACGCCGGAACCTGTCGGT
	GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG
	CAGGTTTC T AGGGCGTG	1927
	CACGCCCT A GAAACCTG	1928

Waxy starch	CCCCGGAACCCGGCGGATGCGGCCCTCGTCATGAGGACTATCGG	1929
GBSS	AGCGAGCGCCGCCCCG T AGCAAAGCCGGAAAGCGCACCGCGGG
Triticum durum	AGCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA
Lys52Term	TGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGCGGTG	1930
AAG-TAG	CGCTTTCCGGCTTTGCT A CGGGGCGGCGCTCGCTCCGATAGTCCT
	CATGACGAGGGCCGCATCCGCCGGGTTCCGGGG
	CCGCCCCG T AGCAAAGC	1931
	GCTTTGCT A CGGGGCGG	1932

Waxy starch	CGGAACCCGGCGGATGCGGCCCTCGTCATGAGGACTATCGGAGC	1933
GBSS	GAGCGCCGCCCCGAAG T AAAGCCGGAAAGCGCACCGCGGGAGC
Triticum durum CGGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG
Gln53Term	CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGCTCCCGCG	1934
CAA-TAA	GTGCGCTTTCCGGCTTT A CTTCGGGGCGGCGCTCGCTCCGATAGT
	CCTCATGACGAGGGCCGCATCCGCCGGGTTCCG
	CCCCGAAG T AAAGCCGG	1935
	CCGGCTTT A CTTCGGGG	1936

Waxy starch	GCGGATGCGGCCCTCGTCATGAGGACTATCGGAGCGAGCGCCGC	1937
GBSS	CCCGAAGCAAAGCCGG T AAGCGCACCGCGGGAGCCGGCGGTGC
Triticum durum	CTCTCCATGGTGGTGCGCGCCACGGGCAGCGGCG
Lys56Term	CGCCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG	1938
AAA-TAA	GCTCCCGCGGTGCGCTT A CCGGCTTTGCTTCGGGGCGGGGCTCG
	CTCCGATAGTCCTCATGACGAGGGCCGCATCCGC
	AAAGCCGG T AAGCGCAC	1939
	GTGCGCTT A CCGGCTTT	1940

Waxy starch	TATCGGAGCGAGCGCCGCCCCGAAGCAAAGCCGGAAAGCGCACC	1941
GBSS	GCGGGAGCCGGCGGTG A CTCTCCATGGTGGTGCGCGCCACGGG
Triticum durum	CAGCGGCGGCATGAACCTCGTGTTCGTCGGCGCC
Cys64Term	GGCGCCGACGAACACGAGGTTCATGCCGCCGCTGCCCGTGGCGC	1942
TGC-TGA	GCACCACCATGGAGAG T CACCGCCGGCTCCCGCGGTGCGCTTTC
	CGGCTTTGCTTCGGGGCGGCGCTCGCTCCGATA
	CGGCGGTG A CTCTCCAT	1943
	ATGGAGAG T CACCGCCG	1944

Waxy starch	CAGCTCGCCACCTCCGGCACCGTCCTCGGCATCACCGACAGGTT	1945
GBSS	CCGGCGTGCAGGTTTT T AGGGTGTGAGGCCCCGGAGCCCGGCAG
Triticum turgidum	ATGCGCCGCTCGGCATGAGGACTACCGGAGCGA
Gln28Term	TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATCTGCGGGGCTC	1946
CAG-TAG	CGGGGCCTCACACCCT A AAAACGTGCACGCCGGAACCTGTCGGT
	GATGCCGAGGACGGTGCCGGAGGTGGCGAGCTG
	CAGGTTTT T AGGGTGTG	1947
	CACACCCT A AAAACCTG	1948

Waxy starch	CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGG	1949
GBSS	AGCGAGCGCCGCCCCG T AGCAACAAAGCCGGAAAGCGCACCGCG
Triticum turgidum	GGACCCGGCGGTGCCTCTCCATGGTGGTGCGCG
Lys52Term	CGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTGCGC	1950
AAG-TAG	TTTCCGGCTTTGTTGCT A CGGGGCGGCGCTCGCTCCGGTAGTCCT
	CATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG
	CCGCCCCG T AGCAACAA	1951
	TTGTTGCT A CGGGGCGG	1952

Waxy starch	CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAG	1953
GBSS	CGAGCGCCGCCCCGAAG T AACAAAGCCGGAAAGCGCACCGCGG
Triticum turgidum	GACCCGGCGGTGCCTCTCCATGGTGGTGCGCGCCA
Gln53Term	TGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCGGTG	1954
CAA-TAA	CGCTTTCCGGCTTTGTT A CTTCGGGGCGGCGCTCGCTCCGGTAGT
	CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG
	CCCCGAAG T AACAAAGC	1955
	GCTTTGTT A CTTCGGGG	1956

Waxy starch	AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTACCGGAGCGAG	1957
GBSS	CGCCGCCCCGAAGCAA T AAAGCCGGAAAGCGCACCGCGGGACCC
Triticum turgidum	GGCGGTGCCTCTCCATGGTGGTGCGCGCCACGG
Gln54Term	CCGTGGCGCGCACCACCATGGAGAGGCACCGCCGGGTCCCGCG	1958
CAA-TAA	GTGCGCTTTCCGGCTTT A TTGCTTCGGGGCGGCGCTCGCTCCGGT
	AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT
	CGAAGCAA T AAAGCCGG	1959
	CCGGCTTT A TTGCTTCG	1960

Waxy starch	GATGCGCCGCTCGGCATGAGGACTACCGGAGCGAGCGCCGCCCC	1961
GBSS	GAAGCAACAAAGCCGG T AAGCGCACCGCGGGACCCGGCGGTGC
Triticum turgidum	CTCTCCATGGTGGTGCGCGCCACGGGCAGCGCCG
Lys57Term	CGGCGCTGCCCGTGGCGCGCACCACCATGGAGAGGCACCGCCG	1962
AAA-TAA	GGTCCCGCGGTGCGCTT A CCGGCTTTGTTGCTTCGGGGCGGCGC
	TCGCTCCGGTAGTCCTCATGCCGAGCGGCGCATC
	AAAGCCGG T AAGCGCAC	1963
	GTGCGCTT A CCGGCTTT	1964

Waxy starch	CAGCTCGCCACCTCCGCCACCGTCCTCGGCATCACCGACAGGTTC	1965
GBSS	CGCCATGCAGGTTTC T AGGGCGTGAGGCCCCGGAGCCCGGCAGA
Aegilops speltoides	TGCGCCGCTCGGCATGAGGACTGTCGGAGCGA
Gln28Term	TCGCTCCGACAGTCCTCATGCCGAGCGGCGCATCTGCCGGGCTC	1966
CAG-TAG	CGGGGCCTCACGCCCT A GAAACCTGCATGGCGGAACCTGTCGGT
	GATGCCGAGGACGGTGGCGGAGGTGGCGAGCTG
	CAGGTTTC T AGGGCGTG	1967
	CACGCCCT A GAAACCTG	1968

Waxy starch	GGTTTCCAGGGCGTGAGGCCCCGGAGCCCGGCAGATGCGCCGCT	1969
GBSS	CGGCATGAGGACTGTC T GAGCGAGCGCCGCCCCGAAGCAACAAA
Aegilops speltoides	GCCGGAAAGCGCACCGCGGGACCCGGCGGTGCC
Gly46Term	GGCACCGCCGGGTCCCGCGGTGCGCTTTCCGGCTTTGTTGCTTC	1970
GGA-TGA	GGGGCGGCGCTCGCTC A GACAGTCCTCATGCCGAGCGGCGCATC
	TGCCGGGCTCCGGGGCCTCACGCCCTGGAAACC
	GGACTGTC T GAGCGAGC	1971
	GCTCGCTC A GACAGTCC	1972

Waxy starch	CCCCGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGG	1973
GBSS	AGCGAGCGCCGCCCCG T AGCAACAAAGCCGGAAAGCGCACCGCG
Aegilops speltoides	GGACCCGGCGGTGCCTCTCGATGGTGGTGCGCG
Lys52Term	CGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGGTGCGCT	1974
AAG-TAG	TTCCGGCTTTGTTGCT A CGGGGCGGCGCTCGCTCCGACAGTCCTC
	ATGCCGAGCGGCGCATCTGCCGGGCTCCGGGG
	CCGCCCCG T AGCAACAA	1975
	TTGTTGCT A CGGGGCGG	1976

Waxy starch	CGGAGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAG	1977
GBSS	CGAGCGCCGCCCCGAAG T AACAAAGCCGGAAAGCGCACCGCGG
Aegilops speltoides	GACCCGGCGGTGCCTCTCGATGGTGGTGCGCGCCA
Gln53Term	TGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCGGTG	1978
CAA-TAA	CGCTTTCCGGCTTTGTT A CTTCGGGGCGGCGCTCGCTCCGACAGT
	CCTCATGCCGAGCGGCGCATCTGCCGGGCTCCG
	CCCCGAAG T AACAAAGC	1979
	GCTTTGTT A CTTCGGGG	1980

Waxy starch	AGCCCGGCAGATGCGCCGCTCGGCATGAGGACTGTCGGAGCGAG	1981
GBSS	CGCCGCCCCGAAGCAA T AAAGCCGGAAAGCGCACCGCGGGACCC
Aegilops speltoides	GGCGGTGCCTCTCGATGGTGGTGCGCGCCACCG
Gln54Term	CGGTGGCGCGCACCACCATCGAGAGGCACCGCCGGGTCCCGCG	1982
CAA-TAA	GTGCGCTTTCCGGCTTT A TTGCTTCGGGGCGGCGCTCGCTCCGAC
	AGTCCTCATGCCGAGCGGCGCATCTGCCGGGCT
	CGAAGCAA T AAAGCCGG	1983
	CCGGCTTT A TTGCTTCG	1984

Waxy starch	AGTGCAGAGATCTTCCACAGCAACAGCTAGACAACCACCATGTCG	1985
GBSS	GCTCTCACCACGTCC T AGCTCGCCACCTCGGCCACCGGCTTCGG
Oryza glaberrima	CATCGCTGACAGGTCGGCGCCGTCGTCGCTGC
Gln8Term	GCAGCGACGACGGCGCCGACCTGTCAGCGATGCCGAAGCCGGT	1986
GAG-TAG	GGCCGAGGTGGCGAGCT A GGACGTGGTGAGAGCCGACATGGTG
	GTTGTCTAGCTGTTGCTGTGGAAGATCTCTGCACT
	CCACGTCC T AGCTCGCC	1987
	GGCGAGCT A GGACGTGG	1988

Waxy starch	TCCACAGCAACAGCTAGACAACCACCATGTCGGCTCTCACCACGT	1989
GBSS	CCCAGCTCGCCACCT A GGCCACCGGCTTCGGCATCGCTGACAGG
Oryza glaberrima	TCGGCGCCGTCGTCGCTGCTCCGCCACGGGTT
Ser12Term	AACCCGTGGCGGAGCAGCGACGACGGCGCCGACCTGTCAGCGAT	1990
TCG-TAG	GCCGAAGCCGGTGGCC T AGGTGGCGAGCTGGGACGTGGTGAGA
	GCCGACATGGTGGTTGTCTAGCTGTTGCTGTGGA
	CGCCACCT A GGCCACCG	1991
	CGGTGGCC T AGGTGGCG	1992

Waxy starch	CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC	1993
GBSS	GGCATCGCTGACAGGT A GGCGCCGTCGTCGCTGCTCCGCCACGG
Oryza glaberrima	GTTCCAGGGCCTCAAGCCCCGCAGCCCCGCCGG
Ser22Term	CCGGCGGGGCTGCGGGGCTTGAGGCCCTGGAACCCGTGGCGGA	1994
TCG-TAG	GCAGCGACGACGGCGCC T ACCTGTCAGCGATGCCGAAGCCGGTG
	GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG
	TGACAGGT A GGCGCCGT	1995
	ACGGCGCC T ACCTGTCA	1996

Waxy starch	CCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCT	1997
GBSS	GACAGGTCGGCGCCGT A GTCGCTGCTCCGCCACGGGTTCCAGGG
Oryza glaberrima	CCTCAAGCCCCGCAGCCCCGCCGGCGGCGACGC
Ser25Term	GCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGAACC	1998
TCG-TAG	CGTGGCGGAGCAGCGAC T ACGGCGCCGACCTGTCAGCGATGCCG
	AAGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG
	GGCGCCGT A GTCGCTGC	1999
	GCAGCGAC T ACGGCGCC	2000

Waxy starch	CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCTGAC	2001
GBSS	AGGTCGGCGCCGTCGT A GCTGCTCCGCCACGGGTTCCAGGGCCT
Oryza glaberrima	CAAGCCCCGCAGCCCCGCCGGCGGCGACGCGAC
Ser26Term	GTCGCGTCGCCGCCGGCGGGGCTGCGGGGCTTGAGGCCCTGGA	2002
TCG-TAG	ACCCGTGGCGGAGCAGC T ACGACGGCGCCGACCTGTCAGCGATG
	CCGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACG
	GCCGTCGT A GCTGCTCC	2003
	GGAGCAGC T ACGACGGC	2004

Waxy starch	TCCACAGCAAGAGCTAAACAGCCGACCGTGTGCACCACCATGTCG	2005
GBSS	GCTGTCACCACGTCC T AGCTCGCCACCTCGGCCACCGGCTTCGG
Oryza sativa	CATCGCCGACAGGTCGGCGCCGTCGTCGCTGG
Gln8Term	GCAGCGACGACGGCGCCGACCTGTCGGCGATGCCGAAGCCGGT	2006
CAG-TAG	GGCCGAGGTGGCGAGCT A GGACGTGGTGAGAGCCGACATGGTG
	GTGCACACGGTCGGCTGTTTAGCTCTTGCTGTGGA
	CCACGTCC T AGCTCGCC	2007
	GGCGAGCT A GGACGTGG	2008

Waxy starch	CTAAACAGCCGACCGTGTGCACCACCATGTCGGCTCTCACCACGT	2009
GBSS	CCCAGCTCGCCACCT A GGCCACCGGCTTCGGCATCGCCGACAGG
Oryza sativa	TCGGCGCCGTCGTCGCTGCTTCGCCACGGGTT
Ser12Term	AACCCGTGGCGAAGCAGCGACGACGGCGCCGACCTGTCGGCGAT	2010
TCG-TAG	GCCGAAGCCGGTGGCC T AGGTGGCGAGCTGGGACGTGGTGAGA
	GCCGACATGGTGGTGCACACGGTCGGCTGTTTAG
	CGCCACCT A GGCCACCG	2011
	CGGTGGCC T AGGTGGCG	2012

Waxy starch	CGGCTCTCACCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTC	2013
GBSS	GGCATCGCCGACAGGT A GGCGCCGTCGTCGCTGCTTCGCCACGG
Oryza sativa	GTTCCAGGGCCTCAAGCCCCGTAGCCCAGCCGG
Ser22Term	CCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCCGTGGCGAA	2014
TCG-TAG	GGAGCGACGACGGCGCC T ACCTGTCGGCGATGCCGAAGCCGGTG
	GCCGAGGTGGCGAGCTGGGACGTGGTGAGAGCCG
	CGACAGGT A GGCGCCGT	2015
	ACGGCGCC T ACCTGTCG	2016

Waxy starch	CCACGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCC	2017
GBSS	GACAGGTCGGCGCCGT A GTCGCTGCTTCGCCACGGGTTCCAGGG
Oryza sativa	CCTCAAGCCCCGTAGCCCAGCCGGCGGGGACGC
Ser25Term	GCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAACCC	2018
TCG-TAG	GTGGCGAAGCAGCGAC T ACGGCGCCGACCTGTCGGCGATGCCGA
	AGCCGGTGGCCGAGGTGGCGAGCTGGGACGTGG
	GGCGCCGT A GTCGCTGC	2019
	GCAGCGAC T ACGGCGCC	2020

Waxy starch	CGTCCCAGCTCGCCACCTCGGCCACCGGCTTCGGCATCGCCGAC	2021
GBSS	AGGTCGGCGCCGTCGT A GCTGCTTCGCCACGGGTTCCAGGGCCT
Oryza sativa	CAAGCCCCGTAGCCCAGCCGGCGGGGACGCATC
Ser26Term	GATGCGTCCCCGCCGGCTGGGCTACGGGGCTTGAGGCCCTGGAA	2022
TCG-TAG	CCCGTGGCGAAGCAGC T ACGACGGCGCCGACCTGTCGGCGATGC
	CGAAGCCGGTGGCCGAGGTGGCGAGCTGGGACG
	GCCGTCGT A GCTGCTTC	2023
	GAAGCAGC T ACGACGGC	2024

Waxy starch	GTCTCTCACTGCAGGTAGCCACACCCTGTGCGCGGCGCCATGGC	2025
GBSS	GGCTCTGGCCACGTCC T AGCTCGCCACCTCCGGCACCGTCCTCG
Hordeum vulgare	GCGTCACCGACAGATTCCGGCGTCCAGGTTTTC
Gln8Term	GAAAACCTGGACGCCGGAATCTGTCGGTGACGCCGAGGACGGTG	2026
GAG-TAG	CCGGAGGTGGCGAGCT A GGACGTGGCCAGAGCCGGCATGGCGC
	CGCGCACAGGGTGTGGCTACCTGCAGTGAGAGAC
	CCACGTCC T AGCTCGCC	2027
	GGCGAGCT A GGACGTGG	2028

Waxy starch	ATGGCGGCTCTGGCCACGTCCCAGCTCGCCACGTCCGGCACCGT	2029
GBSS	CCTCGGCGTCACCGAC T GATTCCGGCGTCCAGGTTTTGAGGGCCT
Hordeum vulgare	CAGGCCCCGGAACCCGGCGGATGCGGCGCTTG
Arg21Term	CAAGCGCGGCATCCGCCGGGTTCCGGGGCCTGAGGCCGTGAAAA	2030
AGA-TGA	CCTGGACGCCGGAATC A GTCGGTGACGCCGAGGACGGTGCCGG
	AGGTGGCGAGCTGGGACGTGGCCAGAGCCGCCAT
	TCACCGAC T GATTCCGG	2031
	CCGGAATC A GTCGGTGA	2032

Waxy starch	CAGCTCGCCACCTCCGGCACCGTCCTCGGCGTCACCGACAGATT	2033
GBSS	CCGGCGTCCAGGTTTT T AGGGCCTCAGGCCCCGGAACCCGGCGG
Hordeum vulgare	ATGCGGCGCTTGGTCTGAGGACTATCGGAGCAA
Gln28Term	TTGCTCCGATAGTCCTCATACCAAGCGCCGCATCCGCCGGGTTCC	2034
CAG-TAG	GGGGCCTGAGGCCCT A AAAACCTGGACGCCGGAATCTGTCGGTG
	ACGCCGAGGACGGTGCCGGAGGTGGCGAGCTG
	CAGGTTTT T AGGGCCTC	2035
	GAGGCCCT A AAAACCTG	2036

Waxy starch	GGTTTTCAGGGCCTCAGGCCGCGGAACCCGGCGGATGCGGCGCT	2037
GBSS	TGGTATGAGGACTATCTGAGCAAGCGCCGCCCCGAAGCAAAGGC
Hordeum vulgare	GGAAAGCGGACCGCGGGAGCCGGCGGTGCCTCT
Gly46Term	AGAGGCACCGCCGGCTCCCGCGGTGCGCTTTCCGGCTTTGCTTC	2038
GGA-TGA	GGGGCGGCGCTTGCTC A GATAGTCCTCATACCAAGCGCCGCATC
	CGCCGGGTTCCGGGGCCTGAGGCCCTGAAAACC
	GGACTATC T GAGCAAGC	2039
	GCTTGCTC A GATAGTCC	2040

Waxy starch	CCCCGGAACCCGGCGGATGCGGCGCTTGGTATGAGGACTATCGG	2041
GBSS	AGCAAGCGCCGCCCCG T AGCAAAGCCGGAAAGCGCACCGCGGG
Hordeum vulgare	AGCCGGCGGTGCCTCTCCGTGGTGGTGAGCGCCA
Lys52Term	TGGCGCTCACCACCACGGAGAGGCACCGCCGGCTCCCGCGGTGC	2042
AAG-TAG	GCTTTGCGGCTTTGCT A CGGGGCGGCGCTTGCTCCGATAGTCCTC
	ATACCAAGCGCCGCATCCGCCGGGTTCCGGGG
	CCGCCCCG T AGCAAAGC	2043
	GCTTTGCT A CGGGGCGG	2044

Waxy starch	ACGTCTTTTCTCTCTCTCCTACGCAGTGGATTAATCGGCATGGCGG	2045
GBSS	CTCTGGCCACGTCG T AGCTCGTCGCAACGCGGGCCGGCCTGGGC
Zea mays	GTCCCGGACGCGTCCACGTTCCGCCGCGGCG
Gln8Term	CGCCGCGGCGGAACGTGGACGCGTCCGGGACGCCCAGGCCGGC	2046
GAG-TAG	GCGCGTTGCGACGAGCT A CGACGTGGCCAGAGCCGCCATGCCGA
	TTAATCCACTGCGTAGGAGAGAGAGAAAAGACGT
	CCACGTCG T AGCTCGTC	2047
	GACGAGCT A CGACGTGG	2048

Waxy starch	GTCGCAACGCGCGCCGGCCTGGGCGTCCCGGACGCGTCCACGTT	2049
GBSS	CCGCCGCGGCGCCGCG T AGGGCCTGAGGGGGGCCCGGGCGTCG
Zea mays	GCGGGGGCGGACACGCTCAGCATGCGGACCAGCG
Gln30Term	CGCTGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGCCCGG	2050
CAG-TAG	GCCCCCCTCAGGCCCT A CGCGGCGCCGCGGCGGAACGTGGACG
	CGTCCGGGACGCCCAGGCCGGCGCGCGTTGCGAC
	GCGCCGCG T AGGGCCTG	2051
	CAGGCCCT A CGCGGCGC	2052

Waxy starch	TCCCGGACGCGTCCACGTTCCGCCGCGGCGCCGCGCAGGGCCT	2053
GBSS	GAGGGGGGCCCGGGCGT A GGCGGCGGCGGACACGCTCAGCATG
Zea mays	CGGACCAGCGCGCGCGCGGCGCCCAGGCACCAGCA
Ser38Term	TGCTGGTGCCTGGGCGCCGCGCGCGCGCTGGTCCGCATGCTGAG	2054
TCG-TAG	CGTGTCCGCCGCCGCC T ACGCCCGGGCCCCCCTCAGGCCCTGCG
	CGGCGCCGCGGCGGAACGTGGACGCGTCCGGGA
	CCGGGCGT A GGCGGCGG	2055
	CCGCCGCC T ACGCCCGG	2056

Waxy starch	GCGTCGGCGGCGGCGGACACGCTCAGCATGCGGACCAGCGCGC	2057
GBSS	GCGCGGCGCCCAGGCAC T AGCAGCAGGCGCGCCGCGGGGGCAG
Zea mays	GTTCCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCA
Ser57Term	TGCCGGCGCTGGCGCACACGACGAGCGACGGGAACCTGCCCCC	2058
GAG-TAG	GCGGCGCGCCTGCTGCT A GTGCCTGGGCGCCGCGCGCGCGCTG
	GTCCGCATGCTGAGCGTGTCCGCCGCCGCCGACGC
	CCAGGCAC T AGCAGCAG	2059
	CTGCTGCT A GTGCCTGG	2060

Waxy starch	TCGGCGGCGGCGGACACGCTCAGCATGCGGACCAGCGCGCGCG	2061
GBSS	CGGCGCCCAGGCACCAG T AGCAGGCGCGCCGCGGGGGCAGGTT
Zea mays	CCCGTCGCTCGTCGTGTGCGCCAGCGCCGGCATGA
Gln58Term	TCATGCCGGGGCTGGCGCACACGACGAGCGACGGGAACCTGCCC	2062
CAG-TAG	CCGCGGCGCGCCTGCT A CTGGTGCCTGGGCGCCGCGCGCGCGC
	TGGTCCGCATGCTGAGCGTGTCCGCCGCCGCCGA
	GGCACCAG T AGCAGGCG	2063
	CGCCTGCT A CTGGTGCC	2064

EXAMPLE 11

Altering Fatty Acid Content of Plants

Improved means to manipulate fatty acid compositions, from biosynthetic or natural plant sources, are needed. For example, oils containing reduced saturated fatty acids are desired for dietary reasons and oils containing increased saturated fatty acids are also needed as alternatives to current sources of highly saturated oil products, such as tropical oils or chemically hydrogenated oils. It would therefore be advantageous to influence directly the production and composition of fatty acids in crop plants. [0143]
Higher plants synthesize fatty acids, primarily palmitic, stearic and oleic acids, in the plastids (i.e., chloroplasts, proplastids, or other related organelles) as part of the Fatty Acid Synthase (FAS) complex. Fatty acid synthesis is the result of the three enzymatic activities: acyl-ACP elongase, acyl-ACP desaturase and acyl-ACP thioesterases specific for each of palmitoyl-, stearoyl- and oleoyl-ACP. [0144]
A variety of enzymes have been identified that influence the relative levels of saturated vs. unsaturated fatty acids in plants. For example, the enzymes stearoyl-acyl carrier protein (stearoyl-ACP) desaturase, oleoyl desaturase and linoleate desaturase produce unsaturated fatty acids from saturated precursors. Similarly, relative enzymatic activities of the various acyl-ACP thioesterases influences the relative acyl-chain composition of the resultant fatty acids. Consequently a reduction or an increase of the activity of these enzymes can alter the properties of oils produced in a plant. In fact, specific targeting of particular enzymatic activities can results in altered levels of particular fatty acids. [0145]

The attached tables disclose exemplary oligonucleotides base sequences which can be used to generate site-specific mutations in plant genes encoding proteins involved in fatty acid biosynthesis.

TABLE 22


Oligonucleotides to produce plants with reduced palmitate

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Reduced palmitate	TTTGGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGC	2065
Acyl-ACP-thioesterase	CACCTCTGCTACGT A GTCATTCTTTCCTGTACCATCTTCTTCACTTG
Arabidopsis thaliana	ATCCTAATGGAAAAGGCAATAAGATTGG
Ser8Term	CCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATGGTA	2066
TCG-TAG	CAGGAAAGAATGAC T ACGTCGCAGAGGTGGCCACCATGACGAGG
	AGATGAAGCGTTCAAAGACACTGCCACCAAA
	TGCTACGT A GTCATTCT	2067
	AGAATGAC T ACGTAGCA	2068

Reduced palmitate	GGTGGCAGTGTCTTTGAACGCTTCATCTCCTCGTCATGGTGGCCA	2069
Acyl-ACP-thioesterase	CCTCTGCTACGTCGT G ATTCTTTCCTGTACCATCTTCTTCACTTGAT
Arabidopsis thaliana	CCTAATGGAAAAGGCAATAAGATTGGGTC
Ser9Term	GACCCAATCTTATTGCCTTTTCCATTAGGATCAAGTGAAGAAGATG	2070
TCA-TGA	GTACAGGAAAGAAT C ACGACGTAGCAGAGGTGGCCACCATGACG
	AGGAGATGAAGCGTTCAAAGACACTGCCACC
	TACGTCGT G ATTCTTTC	2071
	GAAAGAAT C ACGACGTA	2072

Reduced palmitate	ATCTCCTCGTCATGGTGGCCACCTCTGCTACGTCGTCATTCTTTCC	2073
Acyl-ACP-thioesterase	TGTACCATCTTCTT G ACTTGATCCTAATGGAAAAGGCAATAAGATT
Arabidopsis thaliana	GGGTCTACGAATCTTGCTGGACTCAATTC
Ser17Term	GAATTGAGTCCAGCAAGATTCGTCGACCCAATCTTATTGCCTTTTC	2074
TCA-TGA	CATTAGGATCAAGT C AAGAAGATGGTCCAGGAAAGAATGACGACG
	TAGCAGAGGTGGCCACCATGACGAGGAGAT
	ATCTTCTT G ACTTGATC	2075
	GATCAAGT C AAGAAGAT	2076

Reduced palmitate	GTGGCCACCTCTGCTACGTCGTCATTCTTTCCTGTACCATCTTCTT	2077
Acyl-AGP-thioesterase	CACTTGATCCTAAT T GAAAAGGCAATAAGATTGGGTCTACGAATCT
Arabidopsis thaliana	TGCTGGACTCAATTCTGCACCTAACTCTG
Gly22Term	CAGAGTTAGGTGCAGAATTGAGTCCAGCAAGATTCGTCGACCCAA	2078
GGA-TGA	TCTTATTGCCTTTTC A ATTAGGATCAAGTGAAGAAGATGGTCCAGG
	AAAGAATGACGACGTAGCAGAGGTGGCCAC
	ATCCTAAT T GAAAAGGC	2079
	GCCTTTTC A ATTAGGAT	2080

Reduced palmitate	GCTTGAATTTGTGATCTGATTGGTTAATTGTGGCCACAATGGTTGC	2081
Acyl-ACP-thioesterase	TACTGCCGCCACGT G ATCATTCTTTCCGTTGACTTCCCCTTCTGGG
Garcinia mangostana	GATGCCAAATCGGGCAATCCCGGAAAAGG
Ser8Term	CCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAAGTC	2082
TCA-TGA	AACGGAAAGAATGAT C ACGTGGCGGCAGTAGCAACCATTGTGGCC
	ACAATTAACCAATCAGATCACAAATTCAAGC
	CGCCACGT G ATCATTCT	2083
	AGAATGAT C ACGTGGCG	2084

Reduced palmitate	TGAATTTGTGATCTGATTGGTTAATTGTGGCCACAATGGTTGCTAC	2085
Acyl-ACP-thioesterase	TGCCGCCACGTCAT G ATTCTTTCCGTTGACTTCCCCTTCTGGGGAT
Garcinia mangostana	GCCAAATCGGGCAATCCCGGAAAAGGGTC
Ser9Term	GACCCTTTTCCGGGATTGCCCGATTTGGCATCCCCAGAAGGGGAA	2086
TCA-TGA	GTCAACGGAAAGAAT C ATGACGTGGCGGCAGTAGCAACCATTGTG
	GCCACAATTAACCAATCAGATCACAAATTCA
	CACGTCAT G ATTCTTTC	2087
	GAAAGAAT C ATGACGTG	2088

Reduced palmitate	CTGATTGGTTAATTGTGGCCACAATGGTTGCTACTGCCGCCACGT	2089
Acyl-ACP-thioesterase	CATCATTCTTTCCGT A GACTTCCCCTTCTGGGGATGCCAAATCGGG
Garcinia mangostana	CAATCCCGGAAAAGGGTCGGTGAGTTTTGG
Leu13Term	CCAAAACTCACCGACCCTTTTCCGGGATTGCCCGATTTGGCATCC	2090
TTG-TAG	CCAGAAGGGGAAGTC T ACGGAAAGAATGATGACGTGGCGGCAGT
	AGCAACCATTGTGGCCACAATTAACCAATCAG
	CTTTCCGT A GACTTCCC	2091
	GGGAAGTC T ACGGAAAG	2092

Reduced palmitate	ATGGTTGCTACTGCCGCCACGTCATCATTCTTTCCGTTGACTTCCC	2093
Acyl-ACP-thioesterase	CTTCTGGGGATGCC T AATCGGGCAATCCCGGAAAAGGGTCGGTG
Garcinia mangostana	AGTTTTGGGTCAATGAAGTCGAAATCCGCGG
Lys21Term	CCGCGGATTTCGACTTCATTGACCCAAAACTCACCGACCCTTTTCC	2094
AAA-TAA	GGGATTGCCCGATT A GGCATCCCCAGAAGGGGAAGTCAACGGAA
	AGAATGATGACGTGGCGGCAGTCGCAACCAT
	GGGATGCC T AATCGGGC	2095
	GCCCGATT A GGCATCCC	2096

Reduced palmitate	GGGATTTCAGCACGAAATTGAAGTTGTTTTTAAAAACCATGGTTGC	2097
Acyl-ACP-thioesterase	TACTGCTGTGACAT A GGCGTTTTTCCCAGTCACTTCTTCACCTGAC
Gossypium hirsutum	TCCTCTGACTCGAAAAACAAGAAGCTCGG
Ser8Term	CCGAGCTTCTTGTTTTTCGAGTCAGAGGAGTCAGGTGAAGAAGTG	2098
TCG-TAG	ACTGGGAAAAACGCC T ATGTCACAGCAGTAGCAACCATGGTTTTTA
	AAAACAACTTCAATTTCGTGCTGAAATCCC
	TGTGACAT A GGCGTTTT	2099
	AAAACGCC T ATGTCACA	2100

Reduced palmitate	TGTTTTTAAAAACCATGGTTGCTACTGCTGTGACATCGGCGTTTTT	2101
Acyl-ACP-thioesterase	CCCAGTCACTTCTT G ACCTGACTCCTCTGACTCGAAAAACAAGAAG
Gossypium hirsutum	CTCGGAAGCATCAAGTCGAAGCCATCGGT
Ser16Term	ACCGATGGCTTCGACTTGATGCTTCCGAGCTTCTTGTTTTTCGAGT	2102
TCA-TGA	CAGAGGAGTCAGGT C AAGAAGTGACTGGGAAAAACGCCGATGTCA
	CAGCAGTAGCAACCATGGTTTTTAAAAACA
	CACTTCTT G ACCTGACT	2103
	AGTCAGGT C AAGAAGTG	2104

Reduced palmitate	TTGCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACC	2105
Acyl-ACP-thioesterase	TGACTCCTCTGACT A GAAAAACAAGAAGCTCGGAAGCATCAAGTC
Gossypium hirsutum	GAAGCCATCGGTTTCTTCTGGAAGTTTGCA
Ser22Term	TGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCCG	2106
TCG-TAG	AGCTTCTTGTTTTTC T AGTCAGAGGAGTCAGGTGAAGAAGTGACTG
	GGAAAAACGCCGATGTCACAGCAGTCGCAA
	CTCTGACT A GAAAAACA	2107
	TGTTTTTC T AGTCAGAG	2108

Reduced palmitate	GCTACTGCTGTGACATCGGCGTTTTTCCCAGTCACTTCTTCACCTG	2109
Acyl-ACP-thioesterase	ACTCCTCTGACTCG T AAAACAAGAAGCTCGGAAGCATCAAGTCGA
Gossypium hirsutum	AGCCATCGGTTTGTTCTGGAAGTTTGCAAG
Lys23Term	CTTGCAAACTTCCAGAAGAAACCGATGGCTTCGACTTGATGCTTCC	2110
AAA-TAA	GAGCTTCTTGTTTT A CGAGTCAGAGGAGTCAGGTGAAGAAGTGAC
	TGGGAAAAACGCCGATGTCACAGCAGTAGC
	CTGACTCG T AAAACAAG	2111
	CTTGTTTT A GGAGTCAG	2112

Reduced palmitate	CTCCCGCTCGTTGAAAGACAATGGTGGCTACCGCTGCAAGCTCTG	2113
Acyl-ACP-thioesterase	CATTCTTCCCCGTGT A GTCCCCGGTCACCTCCTCTAGACCAGGAA
Cuphea hookeriana	AGCCCGGAAATGGGTCATCGAGCTTCAGCCC
Ser14Term	GGGCTGAAGCTCGATGACCCATTTCCGGGCTTTCCTGGTCTAGAG	2114
TCG-TAG	GAGGTGACCGGGGAC T ACACGGGGAAGAATGCAGAGCTTGCAGC
	GGTAGCCACCATTGTCTTTCAACGAGCGGGAG
	CCCCGTGT A GTCCCCGG	2115
	CCGGGGAC T ACACGGGG	2116

Reduced palmitate	ATGGTGGCTACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCC	2117
Acyl-ACP-thioesterase	CCGGTCACCTCCTCT T GACCAGGAAAGCCCGGAAATGGGTCATCG
Cuphea hookeriana	AGCTTCAGCCCCATCAAGCCCAAATTTGTCG
Arg21Term	CGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACCCATTTC	2118
AGA-TGA	CGGGCTTTCCTGGTC A AGAGGAGGTGACCGGGGACGACACGGG
	GAAGAATGCAGAGCTTGCAGCGGTAGCCACCAT
	CCTCCTCT T GACCAGGA	2119
	TCCTGGTC A AGAGGAGG	2120

Reduced palmitate	GCTACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTC	2121
Acyl-ACP-thioesterase	ACCTCCTCTAGACCA T GAAAGCCCGGAAATGGGTCATCGAGCTTC
Cuphea hookeriana	AGCCCCATCAAGCCCAAATTTGTCGCCAATG
Gly23Term	CATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATGACC	2122
GGA-TGA	CATTTCCGGGCTTTC A TGGTCTAGAGGAGGTGACCGGGGACGAC
	ACGGGGAAGAATGCAGAGCTTGCAGCGGTAGC
	CTAGACCA T GAAAGCCC	2123
	GGGCTTTC A TGGTCTAG	2124

Reduced palmitate	ACCGCTGCAAGCTCTGCATTCTTCCCCGTGTCGTCCCCGGTCACC	2125
Acyl-ACP-thioesterase	TCCTCTAGACCAGGA T AGCCCGGAAATGGGTCATGGAGCTTCAGC
Cuphea hookeriana	CCCATCAAGCCCAAATTTGTCGCCAATGGCG
Lys24Term	CGCCATTGGCGACAAATTTGGGCTTGATGGGGCTGAAGCTCGATG	2126
AAG-TAG	ACCCATTTCCGGGCT A TCCTGGTCTAGAGGAGGTGACCGGGGAC
	GACACGGGGAAGAATGCAGAGCTTGCAGCGGT
	GACCAGGA T AGCCCGGA	2127
	TCCGGGCT A TCCTGGTC	2128

Reduced palmitate	GCCACCGCTGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGAC	2129
Acyl-ACP-thioesterase	ACCTCCTCTAGGCCG T GAAAGCTGGGAAATGGGTCATCGAGCTTG
Cuphea lanceolata	AGCCCCCTCAAGCCCAAATTTGTCGCCAATG
Gly23Term	CATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGATGACC	2130
GGA-TGA	CATTTCCGAGCTTTC A CGGCCTAGAGGAGGTGTCCGGGGACGGC
	AGGGGGAAGAATGCAGAACTTGCAGCGGTGGC
	CTAGGCCG T GAAAGCTC	2131
	GAGCTTTC A CGGCCTAG	2132

Reduced palmitate	ACCGCTGCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACC	2133
Acyl-ACP-thioesterase	TCCTCTAGGCCGGGA T AGCTCGGAAATGGGTCATCGAGCTTGAGC
Cuphea lanceolata	CCCCTCAAGCCCAAATTTGTCGCCAATGCCG
Lys24Term	CGGCATTGGCGACAAATTTGGGCTTGAGGGGGCTCAAGCTCGAT	2134
AAG-TAG	GACCCATTTCCGAGCT A TCCCGGCCTAGAGGAGGTGTCCGGGGA
	CGGCAGGGGGAAGAATGCAGAACTTGCAGCGGT
	GGCCGGGA T AGCTCGGA	2135
	TCCGAGCT A TCCCGGCC	2136

Reduced palmitate	GCAAGTTCTGCATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCT	2137
Acyl-ACP-thioesterase	AGGCCGGGAAAGCTC T GAAATGGGTCATCGAGCTTGAGCCCCCT
Cuphea lanceolata	CAAGCCCAAATTTGTCGCCAATGCCGGGTTGA
Gly26Term	TCAACCCGGCATTGGCGACAAATTTGGGGTTGAGGGGGCTCAAGC	2138
GGA-TGA	TCGATGACCCATTTC A GAGCTTTCCCGGCCTAGAGGAGGTGTCCG
	GGGACGGCAGGGGGAAGAATGCAGAACTTGC
	GAAAGCTC T GAAATGGG	2139
	CCCATTTC A GAGCTTTC	2140

Reduced palmitate	CATTCTTCCCCCTGCCGTCCCCGGACACCTCCTCTAGGCCGGGAA	2141
Acyl-ACP-thioesterase	AGCTCGGAAATGGGT G ATCGAGCTTGAGCCCCCTCAAGCCCAAAT
Cuphea lanceolata	TTGTCGCCAATGCCGGGTTGAAGGTTAAGGC
Ser29Term	GCCTTAACCTTCAACCCGGCATTGGCGACAAATTTGGGCTTGAGG	2142
TCA-TGA	GGGCTCAAGCTCGAT C ACCCATTTCCGAGCTTTCCCGGCCTAGAG
	GAGGTGTCCGGGGACGGCAGGGGGAAGAATG
	AAATGGGT G ATCGAGCT	2143
	AGCTCGAT C ACCCATTT	2144

Reduced palmitate	CGTTTAAGTGGATCGGACATTTAAGTGTTTTAATCATGGTAGCTAT	2145
Acyl-ACP-thioesterase	GAGTGCTACTGCGT A GCTGTTTCCGGTTTCTTCCCCAAAACCTCAC
Helianthus annuus	TCTGGAGCCAAGACATCTGATAAGCTTGG
Ser9Term	CCAAGCTTATCAGATGTCTTGGCTCCAGAGTGAGGTTTTGGGGAA	2146
TCG-TAG	GAAACCGGAAACAGC T ACGCAGTCGCACTCATAGCTACCATGATT
	AAAACACTTAAATGTCCGATCCACTTAAACG
	TACTGCGT A GCTGTTTC	2147
	GAAACAGC T ACGCAGTA	2148

Reduced palmitate	AGTGTTTTAATCATGGTCGCTATGAGTGCTACTGCGTCGCTGTTTC	2149
Acyl-ACP-thioesterase	CGGTTTCTTCCCCA T AACCTCACTCTGGAGCCAAGACATCTGATAA
Helianthus annuus	GCTTGGAGGTGAACCAGGTAGTGTTGCTG
Lys17Term	CAGCAACACTACCTGGTTCACCTCCAAGCTTATCAGATGTCTTGGC	2150
AAA-TAA	TCCAGAGTGAGGTT A TGGGGAAGAAACCGGAAACAGCGACGCAG
	TAGCACTCATAGCTACCATGATTAAAACACT
	CTTCCCCA T AACCTCAC	2151
	GTGAGGTT A TGGGGAAG	2152

Reduced palmitate	ATGGTAGCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCC	2153
Acyl-ACP-thioesterase	CAAAACCTCACTCT T GAGCCAAGACATCTGATAAGCTTGGAGGTG
Helianthus annuus	AACCAGGTAGTGTTGCTGTGCGCGGAATCA
Gly21Term	TGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAGCTTATC	2154
GGA-TGA	AGATGTCTTGGCTC A AGAGTGAGGTTTTGGGGAAGAAACCGGAAA
	CAGCGACGCAGTAGCACTCATAGCTACCAT
	CTCACTCT T GAGCCAAG	2155
	CTTGGCTC A AGAGTGAG	2156

Reduced palmitate	GCTATGAGTGCTACTGCGTCGCTGTTTCCGGTTTCTTCCCCAAAAC	2157
Acyl-ACP-thioesterase	CTCACTCTGGAGCCT A GACATCTGATAAGCTTGGAGGTGAACCAG
Helianthus annuus	GTAGTGTTGCTGTGCGCGGAATCAAGACAA
Lys23Term	TTGTCTTGATTCCGCGCACAGCAACACTACCTGGTTCACCTCCAAG	2158
AAG-TAG	CTTATCAGATGTCT A GGCTCCAGAGTGAGGTTTTGGGGAAGAAAC
	CGGAAACAGCGACGCAGTCGCACTCATAGC
	CTGGAGCC T AGACATCT	2159
	AGATGTCT A GGCTCCAG	2160

Reduced palmitate	ATGGTGGCTGCTGCAGCAAGTTCTGCATGCTTCCCTGTTCCATCC	2161
Acyl-ACP-thioesterase	CCAGGAGCCTCCCCT T AACCTGGGAAGTTAGGCAACTGGTCATCG
Cuphea palustris	AGTTTGAGCCCTTCCTTGAAGCCCAAGTCAA
Lys21Term	TTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATGACCAGTTGC	2162
AAA-TAA	CTAACTTCCCAGGTT A AGGGGAGGCTCCTGGGGATGGAACAGGG
	AAGCATGCAGAACTTGCTGCAGCAGCCACCAT
	CCTCCCCT T AACCTGGG	2163
	CCCAGGTT A AGGGGAGG	2164

Reduced palmitate	GCTGCAGCAAGTTCTGCATGCTTCCCTGTTCCATCCCCAGGAGCC	2165
Acyl-ACP-thioesterase	TCCCCTAAACCTGGG T AGTTAGGCAACTGGTCATCGAGTTTGAGC
Cuphea palustris	CCTTCCTTGAAGCCCAAGTCAATCCCCAATG
Lys24Term	CATTGGGGATTGACTTGGGCTTCAAGGAAGGGCTCAAACTCGATG	2166
AAG-TAG	ACCAGTTGCCTAACT A CCCAGGTTTAGGGGAGGCTCCTGGGGATG
	GAACAGGGAAGCATGCAGAACTTGCTGCAGC
	AACCTGGG T AGTTAGGC	2167
	GCCTAACT A CCCAGGTT	2168

Reduced palmitate	TGCATGCTTCCCTGTTCCATCCCCAGGAGCCTCCCCTAAACCTGG	2169
Acyl-ACP-thioesterase	GAAGTTAGGCAACTG A TCATCGAGTTTGAGCCCTTCCTTGAAGCC
Cuphea palustris	CAAGTCAATCCCCAATGGCGGATTTCAGGTT
Trp28Term	AACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAAGG	2170
TGG-TGA	GCTCAAACTCGATGA T CAGTTGCCTAACTTCCCAGGTTTAGGGGA
	GGCTCCTGGGGATGGAACAGGGAAGCATGCA
	GGCAACTG A TCATCGAG	2171
	CTCGATGA T CAGTTGCC	2172

Reduced palmitate	CATGCTTCCCTGTTCCATCCCCAGGAGCCTCCCCTAAACCTGGGA	2173
Acyl-ACP-thioesterase	AGTTAGGCAACTGGT G ATCGAGTTTGAGCCCTTCCTTGAAGCCCA
Cuphea palustris	AGTCAATCCCCAATGGCGGATTTCAGGTTAA
Ser29Term	TTAACCTGAAATCCGCCATTGGGGATTGACTTGGGCTTCAAGGAA	2174
TCA-TGA	GGGCTCAAACTCGAT C ACCAGTTGCCTAACTTCCCAGGTTTAGGG
	GAGGCTCCTGGGGATGGAACAGGGAAGCATG
	CAACTGGT G ATCGAGTT	2175
	AACTCGAT C ACCAGTTG	2176

Reduced paimitate	ATGGTGGCTGCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACC	2175
Acyl-ACP-thioesterase	CCGGGAATCTCCCCT T AACCCGGGAAGTTCGGTAATGGTGGCTTT
Cuphea hookeriana	CAGGTTAAGGCAAACGCCAATGCCCATCCTA
Lys21Term	TAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGCCACCATTAC	2178
AAA-TAA	CGAACTTCCCGGGTT A AGGGGAGATTCCCGGGGTTGGAACGGAG
	AAGAATGCAGAACTTGCTGCGGCAGCCACCAT
	TCTCCCCT T AACCCGGG	2179
	CCCGGGTT A AGGGGAGA	2180

Reduced palmitate	GCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACCCCGGGAATC	2181
Acyl-ACP-thioesterase	TCCCCTAAACCCGGG T AGTTCGGTAATGGTGGCTTTCAGGTTAAG
Cuphea hookeriana	GCAAACGCCAATGCCCATCCTAGTCTAAAGT
Lys24Term	ACTTTAGACTAGGATGGGCATTGGCGTTTGCCTTAACCTGAAAGC	2182
AAG-TAG	CACCATTACCGAACT A CCCGGGTTTAGGGGAGATTCCCGGGGTTG
	GAACGGAGAAGAATGCAGAACTTGCTGCGGC
	AACCCGGG T AGTTCGGT	2183
	ACCGAACT A CCCGGGTT	2184

Reduced palmitate	TTCTCCGTTCCAACCCCGGGAATCTCCCCTAAACCCGGGAAGTTC	2185
Acyl-ACP-thioesterase	GGTAATGGTGGCTTT T AGGTTAAGGCAAACGCCAATGCCCATCCT
Cuphea hookeriana	AGTCTAAAGTCTGGCAGCCTCGAGACTGAAG
Gln31Term	CTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGGCATTGG	2186
CAG-TAG	CGTTTGCCTTAACCT A AAAGCCACCATTACCGAACTTCCCGGGTTT
	AGGGGAGATTCCCGGGGTTGGAACGGAGAA
	GTGGCTTT T AGGTTAAG	2187
	CTTAACCT A AAAGCCAC	2188

Reduced palmitate	GTTCCAACCCCGGGAATCTCCCCTAAACCCGGGAAGTTCGGTAAT	2189
Acyl-ACP-thioesterase	GGTGGCTTTCAGGTT T AGGCAAACGCCAATGCCCATCCTAGTCTA
Cuphea hookeriana	AAGTCTGGCAGCCTCGAGACTGAAGATGACA
Lys33Term	TGTCATCTTCAGTCTCGAGGCTGCCAGACTTTAGACTAGGATGGG	2190
AAG-TAG	CATTGGCGTTTGCCT A AACCTGAAAGCCACCATTACCGAACTTCCC
	GGGTTTAGGGGAGATTCCCGGGGTTGGAAC
	TTCAGGTT T AGGCAAAC	2191
	GTTTGCCT A AACCTGAA	2192

Reduced palmitate	ATGTTGAAGCTCTCGTGTAATGCGACTGATAAGTTACAGACCCTCT	2193
Acyl-ACP-thioesterase	TCTCGCATTCTCAT T AACCGGATCCGGCACACCGGAGAACCGTCT
Brassica rapa	CCTCCGTGTCGTGCTCTCATCTGAGGAAAC
Gln21Term	GTTTCCTCAGATGAGAGCACGACACGGAGGAGACGGTTCTCCGGT	2194
CAA-TAA	GTGCCGGATCCGGTT A ATGAGAATGCGAGAAGAGGGTCTGTAACT
	TATCAGTCGCATTACACGAGAGCTTCAACAT
	ATTCTCAT T AACCGGAT	2195
	ATCCGGTT A ATGAGAAT	2196

Reduced palmitate	GCGACTGATAAGTTACAGACCCTCTTCTCGCATTCTCATCAACCGG	2197
Acyl-ACP-thioesterase	ATCCGGCACACCGG T GAACCGTCTCCTCCGTGTCGTGCTCTCATC
Brassica rapa	TGAGGAAACCGGTTCTCGATCCTTTGCGAG
Arg28Term	CTCGCAAAGGATCGAGAACCGGTTTCCTCAGATGAGAGCACGACA	2198
AGA-TGA	CGGAGGAGACGGTTC A CCGGTGTGCCGGATCCGGTTGATGAGAA
	TGCGAGAAGAGGGTCTGTAACTTATCAGTCGC
	CACACCGG T GAACCGTC	2199
	GACGGTTC A CCGGTGTG	2200

Reduced palmitate	CCCTCTTCTCGCATTCTCATCAACCGGATCCGGCACACCGGAGAA	2201
Acyl-ACP-thioesterase	CCGTCTCCTCCGTGT A GTGCTCTCATCTGAGGAAACCGGTTCTCG
Brassica rapa	ATCCTTTGCGAGCGATCGTATCTGCTGATCA
Ser24Term	TGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGGTTTC	2202
TCG-TAG	CTCAGATGAGAGCAC T ACACGGAGGAGACGGTTCTCCGGTGTGC
	CGGATCCGGTTGATGAGAATGCGAGAAGAGGG
	CTCCGTGT A GTGCTCTC	2203
	GAGAGCAC T ACACGGAG	2204

Reduced palmitate	CTTCTCGCATTCTCATCAACCGGATCCGGCACACCGGAGAACCGT	2205
Acyl-ACP-thioesterase	CTCCTCCGTGTCGTG A TCTCATCTGAGGAAACCGGTTCTCGATCC
Brassica rapa	TTTGCGAGCGATCGTATCTGCTGATCAAGGA
Cys25Term	TCCTTGATCAGCAGATACGATCGCTCGCAAAGGATCGAGAACCGG	2206
TGC-TGA	TTTCCTCAGATGAGA T CACGACACGGAGGAGACGGTTCTCCGGTG
	TGCCGGATCCGGTTGATGAGAATGCGAGAAG
	GTGTCGTG A TCTCATCT	2207
	AGATGAGA T CACGACAC	2208

Reduced palmitate	ATTCTTCTTCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGG	2209
Acyl-ACP-thioesterase	GCATCAAAAATGT A GAAGCTTTCGTGTAATGTGACTAACAACTTAC
Brassica napus	ACACCTTCTCCTTCTTCTCCGATTCCTC
Leu2Term	GAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCACA	2210
TTG-TAG	TTACACGAAAGCTTC T ACATTTTTGATGCCCTTTTTTTTTTATGGTTC
	CTGAGGTTTTGGTTTATAGAAGAAGAAT
	AAAAATGT A GAAGCTTT	2211
	AAAGCTTC T ACATTTTT	2212

Reduced palmitate	TCTTCTTCTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGG	2213
Acyl-ACP-thioesterase	CATCAAAAATGTTG T AGCTTTCGTGTAATGTGACTAACAACTTACAC
Brassica napus	ACCTTCTCCTTCTTCTCCGATTCCTCCC
Lys3Term	GGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTTGTTAGTCA	2214
AAG-TAG	CATTACACGAAAGCT A CAACATTTTTGATGCCCTTTTTTTTTTATGG
	TTCCTGAGGTTTTGGTTTATAGAAGAAGA
	AAATGTTG T AGCTTTCG	2215
	CGAAAGCT A CAACATTT	2216

Reduced palmitate	CTATAAACCAAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAA	2217
Acyl-ACP-thioesterase	AATGTTGAAGCTTT A GTGTAATGTGACTAACAACTTACACACCTTCT
Brassica napus	CCTTCTTCTCCGATTCCTCCCTTTTCAT
Ser5Term	ATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTAAGTT	2218
TCG-TAG	GTTAGTCACATTACAC T AAAGCTTCAACATTTTTGATGCCCTTTTTT
	TTTTATGGTTCCTGAGGTTTTGGTTTATAG
	GAAGCTTT A GTGTAATG	2219
	CATTACAC T AAAGCTTC	2220

Reduced palmitate	AAACCAAAACCTCAGGAACCATAAAAAAAAAAGGGCATCAAAAATG	2221
Acyl-ACP-thioesterase	TTGAAGCTTTCGTG A AATGTGACTAACAACTTACACACCTTCTCCTT
Brassica napus	CTTCTCCGATTCCTCCCTTTTCATCCCG
Cys6Term	CGGGATGAAAAGGGAGGAATCGGAGAAGAAGGAGAAGGTGTGTA	2222
TGT-TGA	AGTTGTTAGTCACATT T CACGAAAGCTTCAACATTTTTGATGCCCTT
	TTTTTTTTATGGTTCCTGAGGTTTTGGTTT
	CTTTCGTG A AATGTGAC	2223
	GTCACATT T CACGAAAG	2224

TABLE 23


Oligonucleotides to produce plants with increased stearate

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Increased stearate	GGGAGAGCTCTAGCTCTGTAGAAAAGAAGGATTCATTCATCATATC	2225
stearoyl-ACP	CAGAAATGGCTCTA T AGTTTAACCCTTTGGTGGCATCTCAGCCTTA
desaturase	CAAATTCCCTTCCTCGACTCGTCCGCCAA
Arabidopsis thaliana	TTGGCGGACGAGTCGAGGAAGGGAATTTGTAAGGCTGAGATGCC	2226
Lys4 Term	ACCAAAGGGTTAAACT A TAGAGCCATTTCTGGATATGATGAATGAA
AAG-TAG	TCCTTCTTTTCTACAGAGCTAGAGCTCTCCC
	TGGCTCTA T AGTTTAAC	2227
	GTTAAACT A TAGAGCCA	2228

Increased stearate	CTCTGTAGAAAAGAAGGATTCATTCATCATATCCAGAAATGGCTCT	2229
stearoyl-ACP	AAAGTTTAACCCTT A GGTGGCATCTCAGCCTTACAAATTCCCTTCC
desaturase	TCGACTCGTCCGCCAACTCCTCTTTCAG
Arabidopsis thaliana	CTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAAGGGAATTTGTA	2230
Leu8 Term	AGGCTGAGATGCCACC T AAGGGTTAAACTTTAGAGCCATTTCTGG
TTG-TAG	ATATGATGAATGAATCCTTCTTTTCTACAGAG
	TAACCCTT A GGTGGCAT	2231
	ATGCCACC T AAGGGTTA	2232

Increased stearate	AGAAGGATTCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACC	2233
stearoyl-ACP	CTTTGGTGGCATCT T AGCCTTACAAATTCCCTTCCTCGACTCGTCC
desaturase	GCCAACTCCTTCTTTCAGATCTCCCAAGT
Arabidopsis thaliana	ACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAGTCGAGGAA	2234
Gln12 Term	GGGAATTTGTAAGGCT A AGATGCCACCAAAGGGTTAAACTTTAGA
CAG-TAG	GCCATTTCTGGATATGATGAATGAATCCTTCT
	TGGCATCT T AGCCTTAC	2235
	GTAAGGCT A AGATGCCA	2236

Increased stearate	TCATTCATCATATCCAGAAATGGCTCTAAAGTTTAACCCTTTGGTG	2237
stearoyl-ACP	GCATCTCAGCCTTA G AAATTCCCTTCCTCGACTCGTCCGCCAACTC
desaturase	CTTCTTTCAGATCTCCCAAGTTCCTCTGC
Arabidopsis thaliana	GCAGAGGAACTTGGGAGATCTGAAAGAAGGAGTTGGCGGACGAG	2238
Phe14 Term	TCGAGGAAGGGAATTT C TAAGGCTGAGATGCCACCAAAGGGTTAA
TAC-TAG	ACTTTAGAGCCATTTCTGGATATGATGAATGA
	CAGCCTTA G AAATTCCC	2239
	GGGAATTT C TAAGGCTG	2240

Increased stearate	GAGAGCTCGCTCGTGTCTGAAAGAACATCAAACCTCGTATCAAAAA	2241
stearoyl-ACP	AAAGAAAATGGCAT A GAAGCTTAACCCTTTGGCATCTCAGCCTTAC
desaturase	AAACTCCCTTCCTCGGCTCGTCCGCCAAT
Brassica napus	ATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGATGC	2242
Leu3 Term	CAAAGGGTTAAGCTTC T ATGCCATTTTCTTTTTTTTGATACGAGGTT
TTG-TAG	TGATGTTCTTTCAGACACGAGCGAGCTCTC
	AATGGCAT A GAAGCTTA	2243
	TAAGCTTC T ATGCCATT	2244

Increased stearate	GAGCTCGCTCGTGTCTGAAAGAACATCAAACCTCGTATCAAAAAAA	2245
stearoyl-ACP	AGAAAATGGCATTG T AGCTTAACCCTTTGGCATCTCAGCCTTACAA
desaturase	ACTCCCTTCCTCGGCTCGTCCGCCAATCT
Brassica napus	AGATTGGCGGACGAGCCGAGGAAGGGAGTTTGTAAGGCTGAGAT	2246
Lys4 Term	GCCAAAGGGTTAAGCT A CAATGCCATTTTCTTTTTTTTGATACGAG
AAG-TAG	GTTTGATGTTCTTTCAGACACGAGCGAGCTC
	TGGCATTG T AGCTTAAC	2247
	GTTAAGCT A CAATGCCA	2248

Increased stearate	TCTGAAAGAACATCAAACCTCGTATCAAAAAAAAGAAAATGGCATT	2249
stearoyl-ACP	GAAGCTTAACCCTT A GGCATCTCAGCCTTACAAACTCCCTTCCTCG
desaturase	GCTCGTCCGCCAATCTCTACTCTCAGATC
Brassica napus	GATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAAGGGAGTTT	2250
Leu8 Term	GTAAGGCTGAGATGCC T AAGGGTTAAGCTTCAATGCCATTTTCTTT
TTG-TAG	TTTTTGATACGAGGTTTGATGTTCTTTCAGA
	TAACCCTT A GGCATCTC	2251
	GAGATGCC T AAGGGTTA	2252

Increased stearate	AACATCAAACCTCGTATCAAAAAAAAGAAAATGGCATTGAAGCTTA	2253
stearoyl-ACP	ACCCTTTGGCATCT T AGCCTTACAAACTCCCTTCCTCGGCTCGTCC
desaturase	GCCAATCTCTACTCTCAGATCTCCCAAGT
Brassica napus	ACTTGGGAGATCTGAGAGTAGAGATTGGCGGACGAGCCGAGGAA	2254
Gln11 Term	GGGAGTTTGTAAGGCT A AGATGCCAAAGGGTTAAGCTTCAATGCC
CAG-TAG	ATTTTCTTTTTTTTGATACGAGGTTTGATGTT
	TGGCATCT T AGCCTTAC	2255
	GTAAGGCT A AGATGCCA	2256

Increased stearate	AACCAAAAGAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCA	2257
stearoyl-ACP	ATCCTTTCCTTTCT T AAACCCAAAAGTTACCTTCTTTCGCTCTTCCA
desaturase	CCAATGGCCAGTACCAGATCTCCTAAGT
Ricinus communis	ACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGAAAGAAG	2258
Gln27 Term	GTAACTTTTGGGTTT A AGAAAGGATTGAGCTTGAGAGCCAT
CAA-TAA	TGTTTTTTTTCTTACCTTTTTCTTTTGGTT
	TCCTTTCT T AAACCCAA	2259
	TTGGGTTT A AGAAAGGA	2260

Increased stearate	AAGAAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTT	2261
stearoyl-ACP	TCCTTTCTCAAACC T AAAAGTTACCTTCTTTCGCTCTTCCACCAATG
desaturase	GCCAGTACCAGATCTCCTAAGTTCTACA
Ricinus communis	TGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAGCGA	2262
Gln29 Term	AAGAAGGTAACTTTT A GGTTTGAGAAAGGAAAGGATTGAGCTTGA
CAA-TAA	GAGCCATTGTTTTTTTTCTTACCTTTTTCTT
	CTCAAACC T AAAAGTTA	2263
	TAACTTTT A GGTTTGAG	2264

Increased stearate	AAAAAGGTAAGAAAAAAAACAATGGCTCTCAAGCTCAATCCTTTCC	2265
stearoyl-ACP	TTTCTCAAACCCAA T AGTTACCTTCTTTCGCTCTTCCACCAATGGCC
desaturase	AGTACCAGATCTCCTAAGTTCTACATGG
Ricinus communis	CCATGTAGAACTTAGGAGATCTGGTACTGGCCATTGGTGGAAGAG	2266
Lys30 TermCGAAAGAAGGTAACT A TTGGGTTTGAGAAAGGAAAGGATTGAGCT
AAG-TAG	TGAGAGCCATTGTTTTTTTTCTTACCTTTTT
	AAACCCAA T AGTTACCT	2267
	AGGTAACT A TTGGGTTT	2268

Increased stearate	TCTCAAACCCAAAAGTTACCTTCTTTCGCTCTTCCACCAATGGCCA	2269
stearoyl-ACP	GTACCAGATCTCCT T AGTTCTACATGGCCTCTACCCTCAAGTCTGG
desaturase	TTCTAAGGAAGTTGAGAATCTCAAGAAGC
Ricinus communis	GCTTCTTGAGATTCTCAACTTCCTTAGAACCAGACTTGAGGGTAGA	2270
Lys46 Term	GGCCATGTAGAACT A AGGAGATCTGGTACTGGCCATTGGTGGAG
AAG-TAG	AGCGAAAGAAGGTAACTTTTGGGTTTGAGA
	GATCTCCT T AGTTCTAC	2271
	GTAGAACT A AGGAGATC	2272

Increased stearate	TCTTCTGATTCATTTAATCTTTACTCATCAATGGCTCTGAGACTGAA	2273
stearoyl-ACP	CCCTATCCCCACC T AAACCTTCTCCCTCCCCCAAATGGCCAGTCTC
desaturase	AGATCTCCCAGGTTCCGCATGGCCTCTA
Glycine max	TAGAGGCCATGCGGAACCTGGGAGATCTGAGACTGGCCATTTGG	2274
Gln11 Term	GGGAGGGAGAAGGTTT A GGTGGGGATAGGGTTCAGTCTCAGAGC
CAA-TAA	CATTGATGAGTAAAGATTAAATGAATCAGAAGA
	TCCCCACC T AAACCTTC	2275
	GAAGGTTT A GGTGGGGA	2276

Increased stearate	CTTTACTCATCAATGGCTCTGAGACTGAACCCTATCCCCACCCAAA	2277
stearoyl-ACP	CCTTCTCCCTCCCC T AAATGGCCAGTCTCAGATCTCCCAGGTTCC
desaturase	GCATGGCCTCTACCCTCCGCTCCGGTTCCA
Glycine max	TGGAACCGGAGCGGAGGGTAGAGGCCATGCGGAACCTGGGAGAT	2278
Gln17 Term	CTGAGACTGGCCATTT A GGGGAGGGAGAAGGTTTGGGTGGGGAT
CAA-TAA	AGGGTTCAGTCTCAGAGCCATTGATGAGTAAAG
	CCCTCCCC T AAATGGCC	2279
	GGCCATTT A GGGGAGGG	2280

Increased stearate	GCTCTGAGACTGAACCCTATCCCCACCCAAACCTTCTCCCTCCCC	2281
stearoyl-ACP	CAAATGGCCAGTCTC T GATCTCCCAGGTTCCGCATGGCCTCTACC
desaturase	CTCCGCTCCGGTTCCAAAGAGGTTGAAAATA
Glycine max	TATTTTCAACCTCTTTGGAACCGGAGCGGAGGGTAGAGGCCATGC	2282
Arg22 Term	GGAACCTGGGAGATC A GAGACTGGCCATTTGGGGGAGGGAGAAG
AGA-TGA	GTTTGGGTGGGGATAGGGTTCAGTCTCAGAGC
	CCAGTCTC T GATCTCCC	2283
	GGGAGATC A GAGACTGG	2284

Increased stearate	CAAATGGCCAGTCTCAGATCTCCCAGGTTCCGCATGGCCTCTACC	2285
stearoyl-ACP	CTCCGCTCCGGTTCC T AAGAGGTTGAAAATATTAAGAAGCCATTCA
desaturase	CTCCTCCCAGAGAAGTGCATGTTCAAGTAA
Glycine max	TTACTTGAACATGCACTTCTCTGGGAGGAGTGAATGGCTTCTTAAT	2286
Lys37 Term	ATTTTCAACCTCTT A GGAACCGGAGCGGAGGGTAGAGGCCATGCG
AAA-TAA	GAACCTGGGAGATCTGAGACTGGCCATTTG
	CCGGTTCC T AAGAGGTT	2287
	AACCTCTT A GGAACCGG	2288

Increased stearate	CAACAAGCACACACAAGAACAACATCAACAATGGCGATTCGCATC	2289
stearoyl-ACP	AATACGGCGACGTTT T AATCAGACCTGTACCGTTCATTCGCGTTTC
desaturase	CTCAACCGAAACCTCTCAGATCTCCCAAAT
Helianthus annuus	ATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAATGAAC	2290
Gln11 Term	GGTACAGGTCTGATT A AAACGTCGCCGTATTGATGCGAATCGCCA
CAA-TAA	TTGTTGATGTTGTTCTTGTGTGTGCTTGTTG
	CGACGTTT T AATCAGAC	2291
	GTCTGATT A AAACGTCG	2292

Increased stearate	AAGCACACACAAGAAGCAACATCAACAATGGCGATTCGCATCAATAC	2293
stearoyl-ACP	GGCGACGTTTCAAT G AGACCTGTACCGTTCATTCGCGTTTCCTCAA
desaturase	CCGAAACCTCTCAGATCTCCCAAATTCGC
Helianthus annuus	GCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGGAAACGCGAAT	2294
Ser12 Term	GAACGGTACAGGTCT C ATTGAAACGTCGCCGTATTGATGCGAATC
TCA-TGA	GCCATTGTTGATGTTGTTCTTGTGTGTGCTT
	GTTTCAAT G AGACCTGT	2295
	ACAGGTCT C ATTGAAAC	2296

Increased stearate	AAGAACAACATCAACAATGGCGATTCGCATCAATACGGCGACGTTT	2297
stearoyl-ACP	CAATCAGACCTGTA G CGTTCATTCGCGTTTCCTCAACCGAAACCTC
desaturase	TCAGATCTCCCAAATTCGCCATGGCTTCC
Helianthus annuus	GGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGTTGAGG	2298
Tyr15 Term	AAACGCGAATGAACG C TACAGGTCTGATTGAAACGTCGCCGTATT
TAC-TAG	GATGCGAATCGCCATTGTTGATGTTGTTCTT
	GACCTGTA G CGTTCATT	2299
	AATGAACG C TACAGGTC	2300

Increased stearate	CAACATCAACAATGGCGATTCGCATCAATACGGCGACGTTTCAATC	2301
stearoyl-ACP	AGACCTGTACCGTT G ATTCGCGTTTCCTCAACCGAAACCTCTCAGA
desaturase	TCTCCCAAATTCGCCATGGCTTCCACCAT
Helianthus annuus	ATGGTGGAAGCCATGGCGAATTTGGGAGATCTGAGAGGTTTCGGT	2302
Ser17 Term	TGAGGAAACGCGAAT C AACGGTACAGGTCTGATTGAAACGTCGCC
TCA-TGA	GTATTGATGCGAATCGCCATTGTTGATGTTG
	GTACCGTT G ATTCGCGT	2303
	ACGCGAAT C AACGGTAC	2304

Increased stearate	ACACACAACACACACTCAATCACACACACATCATCATCTTCTTCATC	2305
stearoyl-ACP	AACGATGGCGCTT T GAATGAGTCCGGTGACGCTTCAACGGGAGAT
desaturase	ATATCCTTCATACACTTTTCATCAATCGA
Helianthus annuus	TCGATTGATGAAAAGTGTATGAAGGATATATCTCCCGTTGAAGCGT	2306
Arg4 Term	CACCGGACTCATTC A AAGCGCCATCGTTGATGAAGAAGATGATGA
CGA-TGA	TGTGTGTGTGATTGAGTGTGTGTTGTGTGT
	TGGCGCTT T GAATGAGT	2307
	ACTCATTC A AAGCGCCA	2308

Increased stearate	ACACACACATCATCATCTTCTTCATCAACGATGGCGCTTCGAATGA	2309
stearoyl-ACP	GTCCGGTGACGCTT T AACGGGAGATATATCCTTCATACACTTTTCA
desaturase	TCAATCGAAAAATCTCAGATCTCCTAAAT
Helianthus annuus	ATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTATGAAGG	2310
Gln11 Term	ATATATCTCCCGTT A AAGCGTCACCGGACTCATTCGAAGCGCCATC
CAA-TAA	GTTGATGAAGAAGATGATGATGTGTGTGT
	TGACGCTT T AACGGGAG	2311
	CTCCCGTT A AAGCGTCA	2312

Increased stearate	ACATCATCATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGG	2313
stearoyl-ACP	TGACGCTTCAACGG T AGATATATCCTTCATACACTTTTCATCAATCG
desaturase	AAAAATCTCAGATCTCCTAAATTCGCGA
Helianthus annuus	TCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGAAAAGTGTA	2314
Glu13 Term	TGAAGGATATATCT A CCGTTGAAGCGTCACCGGACTCATTCGAAG
GAG-TAG	CGCCATCGTTGATGAAGAAGATGATGATGT
	TTCAACGG T AGATATAT	2315
	ATATATCT A CCGTTGAA	2316

Increased stearate	ATCTTCTTCATCAACGATGGCGCTTCGAATGAGTCCGGTGACGCTT	2317
stearoyl-ACP	CAACGGGAGATATA G CCTTCATACACTTTTCATCAATCGAAAAATC
desaturase	TCAGATCTCCTAAATTCGCGATGGCTTCC
Helianthus annuus	GGAAGCCATCGCGAATTTAGGAGATCTGAGATTTTTCGATTGATGA	2318
Tyr15 Term	AAAGTGTATGAAGG C TATATCTCCCGTTGAAGCGTCACCGGACTC
TAT-TAG	ATTCGAAGCGCCATCGTTGATGAAGAAGAT
	GAGATATA G CCTTCATA	2319
	TATGAAGG C TATATCTC	2320

Increased stearate	AACTCAGCCAGCTTGCCCCCAAACAACAGCGCAGAAAAACCTTCA	2321
stearoyl-ACP	ACAACAATGGCTCTC T AGCTCAACCCAGTCACCACCTTCCCTTCAA
desaturase	CACGCTCCCTCAACAACTTCTCCTCCAGAT
Linum usitatissimum	ATCTGGAGGAGAAGTTGTTGAGGGAGCGTGTTGAAGGGAAGGTG	2322
Lys4 Term	GTGACTGGGTTGAGCT A GAGAGCCATTGTTGTTGAAGGTTTTTCT
AAG-TAG	GCGCTGTTGTTTGGGGGCAAGCTGGCTGAGTT
	TGGCTCTC T AGCTCAAC	2323
	GTTGAGCT A GAGAGCCA	2324

Increased stearate	GCGCAGAAAAACCTTCAACAACAATGGCTCTCAAGCTCAACCCAG	2325
stearoyl-ACP	TCACCACCTTCCCTT G AACACGCTCCCTCAACAACTTCTCCTCCAG
desaturase	ATCTCCTCGCACCTTTCTCATGGCTGCTTC
Linum usitatissimum	GAAGCAGCCATGAGAAAGGTGCGAGGAGATCTGGAGGAGAAGTT	2326
Ser13 Term	GTTGAGGGAGCGTGTT C AAGGGAAGGTGGTGACTGGGTTGAGCT
TCA-TGA	TGAGAGCCATTGTTGTTGAAGGTTTTTCTGCGC
	CTTCCCTT G AACACGCT	2327
	AGCGTGTT C AAGGGAAG	2328

Increased stearate	CTCAAGCTCAACCCAGTCACCACCTTCCCTTCAACACGCTCCCTCA	2329
stearoyl-ACP	ACAACTTCTCCTCC T GATCTCCTCGCACCTTTCTCATGGCTGCTTC
desaturase	CACTTTCAATTCCACCTCCACCAAGTAAG
Linum usitatissimum	CTTACTTGGTGGAGGTGGAATTGAAAGTGGAAGCAGCCATGAGAA	2330
Arg23 Term	AGGTGCGAGGAGATC A GGAGGAGAAGTTGTTGAGGGAGCGTGTT
AGA-TGA	GAAGGGAAGGTGGTGACTGGGTTGAGCTTGAG
	TCTCCTCC T GATCTCCT	2331
	AGGAGATC A GGAGGAGA	2332

Increased stearate	TCCTCCAGATCTCCTCGCACCTTTCTCATGGCTGCTTCCACTTTCA	2333
stearoyl-ACP	ATTCCACCTCCACC T AGTAAGCATCTCCTCCTCCTCGGAATCTCCG
desaturase	CCGATTTCTTTTAAGCGATTGATCGTAGA
Linum usitatissimum	TCTACGATCAATCGCTTAAAAGAAATCGGCGGAGATTCCGAGGAG	2334
Lys411 Term	GAGGAGATGCTTACT A GGTGGAGGTGGAATTGAAAGTGGAAGCA
AAG-TAG	GCCATGAGAAAGGTGCGAGGAGATCTGGAGGA
	CCTCCACC T AGTAAGCA	2335
	TGCTTACT A GGTGGAGG	2336

Increased stearate	ATGGCACTGAAACTTTGCTTTCCACCCCACAAGATGCCTTCCTTCC	2337
stearoyl-ACP	CCGATGCTCGTATC T GATCTCACAGGGTTTTCATGGCTTCAACTAT
desaturase	TCATTCTCCTTCTATGGAGGTCGGAAAAG
Olea europaeap	CTTTCCGACCTCCATAGAAGGAGAATGAATAGTTGAAGCCATGAA	2338
Arg21 Term	AACCCTGTGAGATC A GATACGAGCATCGGGGAAGGAAGGCATCTT
AGA-TGA	GTGGGGTGGAAAGCAAAGTTTCAGTGCCAT
	CTCGTATC T GATCTCAC	2339
	GTGAGATC A GATACGAG	2340

Increased stearate	CCCACAAGATGCCTTCCTTCCCCGATGCTCGTATCAGATCTCACAG	2341
stearoyl-ACP	GGTTTTCATGGCTT G AACTATTCATTCTCCTTCTATGGAGGTCGGA
desaturase	AAAGTTAAAAAGCCTTTCACGCCTCCACG
Olea europaeap	CGTGGAGGCGTGAAAGGCTTTTTAACTTTTCCGACCTCCATAGAA	2342
Ser29 Term	GGAGAATGAATAGTT C AAGCCATGAAAACCCTGTGAGATCTGATAC
TCA-TGA	GAGCATCGGGGAAGGAAGGCATCTTGTGGG
	CATGGCTT G AACTATTC	2343
	GAATAGTT C AAGCCATG	2344

Increased stearate	GATGCTCGTATCAGATCTCACAGGGTTTTCATGGCTTCAACTATTC	2345
stearoyl-ACP	ATTCTCCTTCTATG T AGGTCGGAAAAGTTAAAAAGCCTTTCACGCC
desaturase	TCCACGAGAGGTACATGTTCAAGTAACCC
Olea europaeap	GGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAGGCTTTT	2346
Glu37 Term	TAACTTTTCCGACCT A CATGAAGGAGAATGAATAGTTGAAGCCAT
GAG-TAG	GAAAACCCTGTGAGATCTGATACGAGCATC
	CTTCTATG T AGGTCGGA	2347
	TCCGACCT A CATAGAAG	2348

Increased stearate	CGTATCAGATCTCACAGGGTTTTCATGGCTTCAACTATTCATTCTC	2349
stearoyl-ACP	CTTCTATGGAGGTC T GAAAAGTTAAAAAGCCTTTCACGCCTCCACG
desaturase	AGAGGTACATGTTCAAGTAACCCATTCCT
Olea europaeap	AGGAATGGGTTACTTGAACATGTACCTCTCGTGGAGGCGTGAAAG	2350
Gly39 Term	GCTTTTTAACTTTTC A GACCTCCATAGAAGGAGAATGAATAGTTGA
GGA-TGA	AGCCATGAAAACCCTGTGAGATCTGATACG
	TGGAGGTC T GAAAAGTT	2351
	AACTTTTC A GACCTCCA	2352

Increased stearate	TTCTCGTTTTTGTCGTCCCCTCTGCTCTCTCTCTCTATCAGGCACG	2353
stearoyl-ACP	GAGAAATGGCACTG T AACTCAGTCCAGTCATGTTTCAATCTCAGAA
desaturase	GCTTCCATTTCTTGCCTCCTATCCGCCTT
Persea americana	AAGGCGGATAGGAGGCAAGAAATGGAAGCTTCTGAGATTGAAACA	2354
Lys4 Term	TGACTGGACTGAGTT A CAGTGCCATTTCTCCGTGCCTGATAGAGA
AAA-TAA	GAGAGAGCAGAGGGGACGACAAAAACGAGAA
	TGGCACTG T AACTCAGT	2355
	ACTGAGTT A CAGTGCCA	2356

Increased stearate	CTGCTCTCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCA	2357
stearoyl-ACP	GTCCAGTCATGTTT T AATCTCAGAAGCTTCCATTTCTTGCCTCCTAT
desaturase	CCGCCTTCCAATCTCAGATCTCCGAGGG
Persea americana	CCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAAGAAAT	2358
Gln11 Term	GGAAGCTTCTGAGATT A AAACATGACTGGACTGAGTTTCAGTGCC
CAA-TAA	ATTTCTCCGTGCCTGATAGAGAGAGAGAGCAG
	TCATGTTT T AATCTCAG	2359
	CTGAGATT A AAACATGA	2360

Increased stearate	TCTCTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCA	2361
stearoyl-ACP	GTCATGTTTCAATCT T AGAAGCTTCCATTTCTTGCCTCCTATCCGCC
desaturase	TTCCAATCTCAGATCTCCGAGGGTTTTCA
Persea americana	TGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAGGCAA	2362
Gln13 Term	GAAATGGAAGCTTCT A AGATTGAAACATGACTGGACTGAGTTTCAG
CAG-TAG	TGCCATTTCTCCGTGCCTGATAGAGAGAGA
	TTCAATCT T AGAAGCTT	2363
	AAGCTTCT A AGATTGAA	2364

Increased stearate	CTCTCTATCAGGCACGGAGAAATGGCACTGAAACTCAGTCCAGTC	2365
stearoyl-ACP	ATGTTTCAATCTCAG T AGCTTCCATTTCTTGCCTCCTATCCGCCTTC
desaturase	CAATCTCAGATCTCCGAGGGTTTTCATGG
Persea americana	CCATGAAAACCCTCGGAGATCTGAGATTGGAAGGCGGATAGGAG	2366
Lys14 Term	GCAAGAAATGGAAGCT A CTGAGATTGAAACATGACTGGACTGAGT
AAG-TAG	TTCAGTGCCATTTCTCCGTGCCTGATAGAGAG
	AATCTCAG T AGCTTCCA	2367
	TGGAAGCT A CTGAGATT	2368

Increased stearate	CCCCGAGATCTCGCTGCCGCTGCTCATGGCGTTCGCGGCGTCCC	2369
stearoyl-ACP	ACACCGCATCGCCGTA G TCCTGCGGCGGCGTGGCGCAGAGGAG
desaturase	GAGCAATGGGATGTCGAAGATGGTGGCCATGGCC
Oryza sativa	GGCCATGGCCACCATCTTCGACATCCCATTGCTCCTCCTCTGCGC	2370
Tyr12 Term	CACGCCGCCGCAGGA C TACGGCGATGCGGTGTGGGACGCCGCG
TAC-TAG	AACGCCATGAGCAGCGGCAGCGAGATCTCGGGG
	TCGCCGTA G TCCTGCGG	2371
	CCGCAGGA C TACGGCGA	2372

Increased stearate	CTGCTCATGGCGTTCGCGGCGTCCCACACCGCATCGCCGTACTCC	2373
stearoyl-ACP	TGCGGCGGCGTGGCG T AGAGGAGGAGCAATGGGATGTCGAAGAT
desaturase	GGTGGCCATGGCCTCCACCATCAACAGGGTCA
Oryza sativa	TGACCCTGTTGATGGTGGAGGCCATGGCCACCATCTTCGACATCC	2374
Gln19 Term	CATTGCTCCTCCTCT A CGCCACGCCGCCGCAGGAGTACGGCGAT
CAG-TAG	GCGGTGTGGGACGCCGCGAACGCCATGAGCAG
	GCGTGGCG T AGAGGAGG	2375
	CCTCCTCT A CGCCACGC	2376

Increased stearate	CCCACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGG	2377
stearoyl-ACP	AGGAGCAATGGGATGT A GAAGATGGTGGCCATGGCCTCCACCAT
desaturase	CAACAGGGTCAAGACTGCTAAGAAGCCCTACAC
Oryza sativa	GTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGGCC	2378
Ser26 Term	ATGGCCACCATCTTC T ACATCCCATTGCTCCTCCTCTGCGCCACGC
TCG-TAG	CGCCGCAGGAGTACGGCGATGCGGTGTGGG
	TGGGATGT A GAAGATGG	2379
	CCATCTTC T ACATCCCA	2380

Increased stearate	CACACCGCATCGCCGTACTCCTGCGGCGGCGTGGCGCAGAGGAG	2381
stearoyl-ACP	GAGCAATGGGATGTCG T AGATGGTGGCCATGGCCTCCACCATCAA
desaturase	CAGGGTCAAGACTGCTAAGAAGCCCTACACTC
Oryza sativa	GAGTGTAGGGCTTCTTAGCAGTCTTGACCCTGTTGATGGTGGAGG	2382
Lys27 Term	CCATGGCCACCATCT A CGACATCCCATTGCTCCTCCTCTGCGCCA
AAG-TAG	CGCCGCCGCAGGAGTACGGCGATGCGGTGTG
	GGATGTCG T AGATGGTG	2383
	CACCATCT A CGACATCC	2384

Increased stearate	TTCTCTCTCTAGGTTGAGCGGTTACCAACAGAAGCACTTAGGAGA	2385
stearoyl-ACP	GAGAAGCAATGGCGT A GAAGCTTCACCACACGGCCTTCAATCCTT
desaturase	CCATGGCGGTTACCTCTTCGGGACTTCCTCG
Simmondsia chinensis	CGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAGGC	2386
Leu3 Term	CGTGTGGTGAAGCTTC T ACGCCATTGCTTCTCTCTCCTAAGTGCTT
TTG-TAG	CTGTTGGTAACCGCTCAACCTAGAGAGAGAA
	AATGGCGT A GAAGCTTC	2387
	GAAGCTTC T ACGCCATT	2388

Increased stearate	CTCTCTCTAGGTTGAGCGGTTACCAACAGAAGCACTTAGGAGAGA	2389
stearoyl-ACP	GAGCAATGGCGTTG T AGCTTCACCACACGGCCTTCAATCCTTCC
desaturase	ATGGCGGTTACCTCTTCGGGACTTCCTCGAT
Simmondsia chinensis	ATCGAGGAAGTCCCGAAGAGGTAACCGCCATGGAAGGATTGAAG	2390
Lys4 Term	GCCGTGTGGTGAAGCT A CAACGCCATTGCTTCTCTCTCCTAAGTG
AAG-TAG	CTTCTGTTGGTAACCGCTCAACCTAGAGAGAG
	TGGCGTTG T AGCTTCAC	2391
	GTGAAGCT A CAACGCCA	2392

Increased stearate	AAGCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCAT	2393
stearoyl-ACP	GGCGGTTACCTCTT A GGGACTTCCTCGATCGTATCACCTCAGATCT
desaturase	CACCGCGTTTTCATGGCTTCTTCTACAAT
Simmondsia chinensis	ATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGATAC	2394
Ser19 Term	GATCGAGGAAGTCCC T AAGAGGTAACCGCCATGGAAGGATTGAAG
TCG-TAG	GCCGTGTGGTGAAGCTTCAACGCCATTGCTT
	TACCTCTT A GGGACTTC	2395
	GAAGTCCC T AAGAGGTA	2396

Increased stearate	GCAATGGCGTTGAAGCTTCACCACACGGCCTTCAATCCTTCCATG	2397
stearoyl-ACP	GCGGTTACCTCTTCG T GACTTCCTCGATCGTATCACCTCAGATCTC
desaturase	ACCGCGTTTTCATGGCTTCTTCTACAATTG
Simmondsia chinensis	CAATTGTAGAAGAAGCCATGAAAACGCGGTGAGATCTGAGGTGAT	2398
Gly20 Term	ACGATCGAGGAAGTC A CGAAGAGGTAACCGCCATGGAAGGATTG
GGA-TGA	AAGGCCGTGTGGTGAAGCTTCAACGCCATTGC
	CCTCTTCG T GACTTCCT	2399
	AGGAAGTC A CGAAGAGG	2400

Increased stearate	TGGCTCTGAATCTCAACCCCGTTTCCACACCATTTCAGTGTCGTCG	2401
stearoyl-ACP	ATTGCCGTCTTTCT G ACCTCGTCAAACGCCTTCTCGCAGATCTCCC
desaturase	AAATTCTTCATGGCTTCCACTCTCAGCAG
Spinacia oleracea	CTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATCTGCGAGAA	2402
Ser21 Term	GGCGTTTGACGAGGT C AGAAAGACGGCAATCGACGACACTGAAAT
TCA-TGA	GGTGTGGAAACGGGGTTGAGATTCAGAGCCA
	GTCTTTCT G ACCTCGTC	2403
	GACGAGGT C AGAAAGAC	2404

Increased stearate	AATCTCAACCCCGTTTCCACACCATTTCAGTGTCGTCGATTGCCGT	2405
stearoyl-ACP	CTTTCTCACCTCGT T AAACGCCTTCTCGCAGATCTCCCAAATTCTT
desaturase	CATGGCTTCCACTCTCAGCAGCTCTTCTC
Spinacia oleracea	GAGAAGAGCTGCTGAGAGTGGAAGCCATGAAGAATTTGGGAGATC	2406
Gln24 Term	TGCGAGAAGGCGTTT A ACGAGGTGAGAAAGACGGCAATCGACGA
CAA-TAA	CACTGAAATGGTGTGGAAACGGGGTTGAGATT
	CACCTCGT T AAACGCCT	2407
	AGGCGTTT A ACGAGGTG	2408

Increased stearate	TCCACACCATTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTC	2409
stearoyl-ACP	AAACGCCTTCTCGC T GATCTCCCAAATTCTTCATGGCTTCCACTCT
desaturase	CAGCAGCTCTTCTCCTAAGGAAGCGGAAA
Spinacia oleracea	TTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGGAAGCCATGA	2410
Arg29 Term	AGAATTTGGGAGATC A GCGAGAAGGCGTTTGACGAGGTGAGAAA
AGA-TGA	GACGGCAATCGACGACACTGAAATGGTGTGGA
	CTTCTCGC T GATCTCCC	2411
	GGGAGATC A GCGAGAAG	2412

Increased stearate	TTTCAGTGTCGTCGATTGCCGTCTTTCTCACCTCGTCAAACGCCTT	2413
stearoyl-ACP	CTCGCAGATCTCCC T AATTCTTCATGGCTTCCACTCTCAGCAGCTC
desaturase	TTCTCCTAAGGAAGCGGAAAGCCTGAAGA
Spinacia oleracea	TCTTCAGGCTTTCCGCTTCCTTAGGAGAAGAGCTGCTGAGAGTGG	2414
Lys32 Term	AAGCCATGAAGAATT A GGGAGATCTGCGAGAAGGCGTTTGACGAG
AAA-TAA	GTGAGAAAGACGGCAATCGACGACACTGAAA
	GATCTCCC T AATTCTTC	2415
	GAAGAATT A GGGAGATC	2416

Increased stearate	AAATAGTCGAGGTGAAAAACAGAGCATCAACAATGGCACTGAATAT	2417
stearoyl-ACP	CAATGGGGTGTCGT G AAAATCTCACAAAATGTTACCATTTCCTTGT
desaturase	TCTTCAGCCAGATCTGAGCGAGTTTTCAT
Solanum tuberosum	ATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTAAC	2418
Leu10 Term	ATTTTGTGAGATTTT C ACGACACCCCATTGATATTCAGTGCCATTGT
TTA-TGA	TGATGCTCTGTTTTTCACCTCGACTATTT
	GGTGTCGT G AAAATCTC	2419
	GAGATTTT C ACGACACC	2420

Increased stearate	ATAGTCGAGGTGAAAACAGAGCATCAACAATGGCACTGAATATCA	2421
stearoyl-ACP	ATGGGGTGTCGTTA T AATCTCACAAAATGTTACCATTTCCTTGTTCT
desaturase	TCAGCCAGATCTGAGCGAGTTTTCATGG
Solanum tuberosum	CCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAGGAAATGGTA	2422
Lys11 Term	ACATTTTGTGAGATT A TAACGACACCCCATTGATATTCAGTGCCATT
AAA-TAA	GTTGATGCTCTGTTTTTCACCTCGACTAT
	TGTCGTTA T AATCTCAC	2423
	GTGAGATT A TAACGACA	2424

Increased stearate	GTGAAAAACAGAGCATCAACAATGGCACTGAATATCAATGGGGTG	2425
stearoyl-ACP	TCGTTAAAATCTCAC T AAATGTTACCATTTCCTTGTTCTTCAGCCAG
desaturase	ATCTGAGCGAGTTTTCATGGCTTCAACCA
Solanum tuberosum	TGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAAGAACAAG	2426
Lys14 Term	GAAATGGTAACATTT A GTGAGATTTTAACGACACCCCATTGATATT
AAA-TAA	CAGTGCCATTGTTGATGCTCTGTTTTTCAC
	AATCTCAC T AAATGTTA	2427
	TAACATTT A GTGAGATT	2428

Increased stearate	ACAGAGCATCAACAATGGCACTGAATATCAATGGGGTGTCGTTAAA	2429
stearoyl-ACP	ATCTCACAAAATGT G ACCATTTCCTTGTTCTTCAGCCAGATCTGAG
desaturase	CGAGTTTTCATGGCTTCAACCATTCATCG
Solanum tuberosum	CGATGAATGGTTGAAGCCATGAAAACTCGCTCAGATCTGGCTGAA	2430
Leu16 Term	GAACAAGGAAATGGT C ACATTTTGTGAGATTTTAACGACACCCCAT
TTA-TGA	TGATATTCAGTGCCATTGTTGATGCTCTGT
	CAAAATGT G ACCATTTC	2431
	GAAATGGT C ACATTTTG	2432

Increased stearate	TGGCTCTGAGGCTGAACCCTAACCCTTCACAGAAGCTCTTTCTCTC	2433
stearoyl-ACP	TCCTTCTTCATCAT G ATCTTCTTCTTCTTCATCGTTCTCGCTTCCTC
desaturase	AAATGGCTAGCCTCAGATCTCCAAGGTT
Arachis hypogaea	AACCTTGGAGATCTGAGGCTAGCCATTTGAGGAAGCGAGAACGAT	2434
Ser21 Term	GAAGAAGAAGAAGAT C ATGATGAAGAAGGAGAGAGAAAGAGCTTC
TCA-TGA	TGTGAAGGGTTAGGGTTCAGCCTCAGAGCCA
	TTCATCAT G ATCTTCTT	2435
	AAGAAGAT C ATGATGAA	2436

Increased stearate	ACCCTAACCCTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCA	2437
stearoyl-ACP	TCTTCTTCTTCTT G ATCGTTCTCGCTTCCTCAAATGGCTAGCCTCA
desaturase	GTCTCCAAGGTTCCGCATGGCCTCCAC
Arachis hypogaea	GTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCATTTGA	2438
Ser26 Term	GGAAGCGAGAACGAT C AAGAAGAAGAAGATGATGATGAAGAAGGA
TCA-TGA	GAGAGAAAGAGCTTCTGTGAAGGGTTAGGGT
	TTCTTCTT G ATCGTTCT	2439
	AGAACGAT C AAGAAGAA	2440

Increased stearate	CTAACCCTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCATCT	2441
stearoyl-ACP	TCTTCTTCTTCAT A GTTCTCGCTTCCTCAAATGGCTAGCCTCAGAT
desaturase	CTCCAAGGTTCCGCATGGCCTCCACCCT
Arachis hypogaea	AGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCTAGCCAT	2442
Ser27 Term	TTGAGGAAGCGAGAAC T ATGAAGAAGAAGAAGATGATGATGAAGA
TCG-TAG	AGGAGAGAGAAAGAGCTTCTGTGAAGGGTTAG
	TTCTTCAT A GTTCTCGC	2443
	GCGAGAAC T ATGAAGAA	2444

Increased stearate	CTTCACAGAAGCTCTTTCTCTCTCCTTCTTCATCATCATCTTCTTCT	2445
stearoyl-ACP	TCTTCATCGTTCT A GCTTCCTCAAATGGCTAGCCTCAGATCTCCAA
desaturase	GGTTCCGCATGGCCTCCACCCTCCGCAC
Arachis hypogaea	GTGCGGAGGGTGGAGGCCATGCGGAACCTTGGAGATCTGAGGCT	2446
Ser29 Term	AGCCATTTGAGGAAGC T AGAACGATGAAGAAGAAGAAGATGATGA
TCG-TAG	TGAAGAAGGAGAGAGAAAGAGCTTCTGTGAAG
	ATCGTTCT A GCTTCCTC	2447
	GAGGAAGC T AGAACGAT	2448

Increased stearate	AAAGTTAAAAGCCGTCCAAAACCCAAACCAGGAAAGGCAAACGAA	2449
stearoyl-ACP	AAGAAAAAATGGCTT A GAATTTTAATGCCATCGCCTCGAAATCTCA
desaturase	GAAGCTCCCTTGCTTTGCTCTTCCACCAAA
Gossypium hirsutum	TTTGGTGGAAGAGCAAAGCAAGGGAGCTTCTGAGATTTCGAGGCG	2450
Leu3 Term	ATGGCATTAAAATTC T AAGCCATTTTTTCTTTTCGTTTGCCTTTCCT
TTG-TAG	GGTTTGGGTTTTGGACGGCTTTTAACTTT
	AATGGCTT A GAATTTTA	2451
	TAAAATTC T AAGCCATT	2452

Increased stearate	CCCAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTT	2453
stearoyl-ACP	TAATGCCATCGCCT A GAAATCTCAGAAGCTCCCTTGCTTTGCTCTT
desaturase	CCACCAAAGGCCACCCTTAGATCTCCCAA
Gossypium hirsutum	TTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAGG	2454
Ser1-Term	GAGCTTCTGAGATTTC T AGGCGATGGCATTAAAATTCAAAGCCATT
TCG-TAG	TTTTCTTTTCGTTTGCCTTTCCTGGTTTGGG
	CATCGCCT A GAAATCTC	2455
	GAGATTTC T AGGCGATG	2456

Increased stearate	CAAACCAGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTA	2457
stearoyl-ACP	ATGCCATCGCCTCG T AATCTCAGAAGCTCCCTTGCTTTGCTCTTCC
desaturase	ACCAAAGGCCACCCTTAGATCTCCCAAGT
Gossypium hirsutum	ACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAAAGCAAG	2458
Lys11 Term	GGAGCTTCTGAGATT A CGAGGCGATGGCATTAAAATTCAAAGCCAA
AAA-TAA	TTTTTTCTTTTCGTTTGCCTTTCCTGGTTTG
	TCGCCTCG T AATCTCAG	2459
	CTGAGATT A CGAGGCGA	2460

Increased stearate	AGGAAAGGCAAACGAAAAGAAAAAATGGCTTTGAATTTTAATGCCA	2461
stearoyl-ACP	TCGCCTCGAAATCT T AGAAGCTCCCTTGCTTTGCTCTTCCACCAAA
desaturase	GGCCACCCTTAGATCTCCCAAGTTTTCCA
Gossypium hirsutum	TGGAAAACTTGGGAGATCTAAGGGTGGCCTTTGGTGGAAGAGCAA	2462
Gln13 Term	AGCAAGGGAGCTTCT A AGATTTCGAGGCGATGGCATTAAAATTCA
CAG-TAG	AAGCCATTTTTTCTTTTCGTTTGCCTTTCCT
	CGAAATCT T AGAAGCTC	2463
	GAGCTTCT A AGATTTCG	2464

TABLE 24


Oligonucleotides to produce plants with reduced linolenic acid

Phenotype, Gene,
Plant & Targeted		SEQ ID
Alteration	Altering Oligos	NO:

Reducing linolenic acid	AATAGAACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGC	2465
omega-3 fatty acid	TCCAATGGCGAGCT A GGTTTTATCAGAATGTGGTTTTAGACCTCTC
desaturase	CCCAGATTCTACCCTAAACACACAACCTC
Arabidopsis thaliana	GAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTAAAACCA	2466
Ser4 Term	CATTCTGATAAAACC T AGCTCGCCATTGGAGCCTCTTCCCAAGAAG
TCG-TAG	AAAAGAGGAAAAAGTCTCTGTCGTTCTATT
	GGCGAGCT T GGTTTTAT	2467
	ATAAAACC A AGCTCGCC	2468

Reducing linolenic acid	ACGACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAAT	2469
omega-3 fatty acid	GGCGAGCTCGGTTT G ATCAGAATGTGGTTTTAGACCTCTCCCCAG
desaturase	ATTCTACCCTAAACACACAACCTCTTTTGC
Arabidopsis thaliana	GCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGTCTA	2470
Leu6 Term	AAACCACATTCTGAT C AAACCGAGCTCGCCATTGGAGCCTCTTCCC
TTA-TGA	AAGAAGAAAAGAGGAAAAAGTCTCTGTCGT
	CTCGGTTT G ATCAGAAT	2471
	ATTCTGAT C AAACCGAG	2472

Reducing linolenic acid	ACAGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGC	2473
omega-3 fatty acid	GAGCTCGGTTTTAT G AGAATGTGGTTTTAGACCTCTCCCCAGATTC
desaturase	TACCCTAAACACACAACCTCTTTTGCCTC
Arabidopsis thaliana	GAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAGGT	2474
Ser7 Term	CTAAAACCACATTCT C ATAAAACCGAGCTCGCCATTGGAGCCTCTT
TCA-TGA	CCAAGAAGAAAAGAGGAAAAAGTCTCTGT
	GGTTTTAT G AGAATGTG	2475
	CACATTCT C ATAAAACC	2476

Reducing linolenic acid	AGAGACTTTTTCCTCTTTTCTTCTTGGGAAGAGGCTCCAATGGCGA	2477
omega-3 fatty acid	GCTCGGTTTTATCA T AATGTGGTTTTAGACCTCTCCCCAGATTCTA
desaturase	CCCTAAACACACAACCTCTTTTGCCTCTA
Arabidopsis thaliana	TAGAGGCAAAAGAGGTTGTGTGTTTAGGGTAGAATCTGGGGAGAG	2478
Glu8 Term	GTCTAAAACCACATT A TGATAAAACCGAGCTCGCCATTGGAGCCTC
GAA-TAA	TTCCCAAGAAGAAAAGAGGAAAAAGTCTCT
	TTTTATCA T AATGTGGT	2479
	ACCACATT A TGATAAAA	2480

Reducing linolenic acid	TCATCATCTTCTTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTC	2481
omega-3 fatty acid	TAGCAATGGCGAACT A GGTCTTATCCGAATGTGGCATAAGACCTC
desaturase	TCCCCAGAATCTACACCACACCCAGATCCAC
Brassica juncea	GTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTTATGCCA	2482
Leu4 Term	CATTCGGATAAGACC T AGTTCGCCATTGCTAGAGCTCTTTTGCTCT
TTG-TAG	CTCTCTCTCCCCAGAAGAAGAAGATGATGA
	GGCGAACT A GGTCTTAT	2483
	ATAAGACC T AGTTCGCC	2484

Reducing linolenic acid	TCTTCTTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAA	2485
omega-3 fatty acid	TGGCGAACTTGGTCT G ATCCGAATGTGGCATAAGACCTCTCCCCA
desaturase	GAATCTACACCACACCCAGATCCACTTTCCT
Brassica juncea	AGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGAGGTCTT	2486
Leu6 Term	ATGCCACATTCGGAT C AGACCAAGTTCGCCATTGCTAGAGCTCTTT
TTA-TGA	TGCTCTCTCTCTCTCCCCAGAAGAAGAAGA
	CTTGGTCT G ATCCGAAT	2487
	ATTCGGAT C AGACCAAG	2488

Reducing linolenic acid	TTCTTCTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCG	2489
omega-3 fatty acid	AACTTGGTCTTATCC T AATGTGGCATAAGACCTCTCCCCAGAATCT
desaturase	ACACCACACCCAGATCCACTTTCCTCTCCA
Brassica juncea	TGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGGGGAGA	2490
Glu8 Term	GGTCTTATGCCACATT A GGATAAGACCAAGTTCGCCATTGCTAGA
GAA-TAA	GCTCTTTTGCTCTCTCTCTCTCCCCAGAAGAA
	TCTTATCC T AATGTGGC	2491
	GCCACATT A GGATAAGA	2492

Reducing linolenic acid	CTGGGGAGAGAGAGAGAGCAAAAGAGCTCTAGCAATGGCGAACT	2493
omega-3 fatty acid	TGGTCTTATCCGAATG A GGCATAAGACCTCTCCCCAGAATCTACAC
desaturase	CACACCCAGATCCACTTTCCTCTCCAACACC
Brassica juncea	GGTGTTGGAGAGGAAAGTGGATCTGGGTGTGGTGTAGATTCTGG	2494
Cys9 Term	GGAGAGGTCTTATGCC T CATTCGGATAAGACCAAGTTCGCCATTG
TGT-TGA	CTAGAGCTCTTTTGCTCTCTCTCTCTCCCCAG
	TCCGAATG A GGCATAAG	2495
	CTTATGCC T CATTCGGA	2496

Reducing linolenic acid	ATAACAGAATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAA	2497
omega-3 fatty acid	TGGCTGCTGGTTG A GTATTATCAGAATGTGGTTTAAGGCCTCTCCC
desaturase	AAGAATCTACTCACGACCCAGAATTGGT
Ricinus communis	ACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTTAAACC	2498
Trp5 Term	ACATTCTGATAATAC T CAACCAGCAGCCATTGAAAACCCAGAAGCT
TGG-TGA	AAAAATGCAAGAATTCAGCAATTCTGTTAT
	GCTGGTTG A GTATTATC	2499
	GATAATAC T CAACCAGC	2500

Reducing linolenic acid	AGAATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCT	2501
omega-3 fatty acid	GCTGGTTGGGTAT G ATCAGAATGTGGTTTAAGGCCTCTCCCAAGA
desaturase	ATCTACTCACGACCCAGAATTGGTTTTAC
Ricinus communis	GTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGCCTT	2502
Leu7 Term	AAACCACATTCTGAT C ATACCCAACCAGCAGCCATTGAAAACCCAG
TTA-TGA	AAGCTAAAAATGCAAGAATTCAGCAATTCT
	TTGGGTAT GATCAGAAT	2503
	ATTCTGAT C ATACCCAA	2504

Reducing linolenic acid	ATTGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCTGCT	2505
omega-3 fatty acid	GGTTGGGTATTAT G AGAATGTGGTTTAAGGCCTCTCCCAAGAATCT
desaturase	ACTCACGACCCAGAATTGGTTTTACATC
Ricinus communis	GATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGGGAGAGGC	2506
Ser8 Term	CTTAAACCACATTCT C ATAATACCCAACCAGCAGCCATTGAAAACC
TCA-TGA	CAGAAGCTAAAAATGCAAGAATTCAGCAAT
	GGTATTAT G AGAATGTG	2507
	CACATTCT C ATAATACC	2508

Reducing linolenic acid	TGCTGAATTCTTGCATTTTTAGCTTCTGGGTTTTCAATGGCTGCTG	2509
omega-3 fatty acid	GTTGGGTATTATCA T AATGTGGTTTAAGGCCTCTCCCAAGAATCTA
desaturase	CTCACGACCCAGAATTGGTTTTACATCGA
Ricinus communis	TCGATGTAAAACCAATTCTGGGTCGTGAGTAGATTCTTGFGGAGAG	2510
Glu9 Term	CGCCTTAAACCACATT A TGATAATACCCAACCAGCAGCCATTGAAAA
GAA-TAA	CCCAGAAGCTAAAAATGCAAGAATTCAGCA
	TATTATCA T AATGTGGT	2511
	ACCACATT A TGATAATA	2512

Reducing linolenic acid	GCAAGTTGGTTTTATCAGAATGTGGTCTTAGACCACTCCCAAGAA	2513
omega-3 fatty acid	TCTACCCTAAGCCC T GAACTGGGGCAGCCACTTCTGCCTCCTCTC
desaturase	ACATTAAGTTGAGAATTTCACGTACAGATC
Nicotiana tabacum	GATCTGTACGTGAAATTCTCAACTTAATGTGAGAGGAGGCAGAAGT	2514
Arg22 Term	GGCTGCCCCAGTTC A GGGCTTAGGGTAGFATTCTTGGGAGTGGTCT
AGA-TGA	AAGACCACATTCTGATAAAACCCAACTTGC
	CTAAGCCC T GAACTGGG	2515
	CCCAGTTC A GGGCTTAG	2516

Reducing linolenic acid	CTCCCAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCT	2517
omega-3 fatty acid	GCCTCCTCTCACATT T AGTTGAGAATTTCACGTACAGATCTGAGTG
desaturase	GTTCTGCAATTTCTTTGTCTAATACTAAT
Nicotiana tabacum	TATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGTACG	2518
Lys34 Term	TGAAATTCTCAACT A AATGTGAGAGGAGGCAGAAGTGGCTGCCCC
AAG-TAG	AGTTCTGGGCTTAGGGTAGATTCTTGGGAG
	CTCACATT T AGTTGAGA	2519
	TCTCAACT A AATGTGAG	2520

Reducing linolenic acid	CAAGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCT	2521
omega-3 fatty acid	CCTCTCACATTAAGT A GAGAATTTCACGTACAGATCTGAGTGGTTC
desaturase	TGCAATTTCTTTGTCTAATACTAATAAAGA
Nicotiana tabacum	TCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCTGT	2522
Leu35 Term	ACGTGAAATTCTC T ACTTAATGTGAGAGGAGGCAGAAGTGGCTGC
TTG-TAG	CCCAGTTCTGGGCTTAGGGTAGATTCTTG
	CATTAAGT A GAGAATTT	2523
	AAATTCTC T ACTTAATG	2524

Reducing linolenic acid	AGAATCTACCCTAAGCCCAGAACTGGGGCAGCCACTTCTGCCTCC	2525
omega-3 fatty acid	TCTCACATTAAGTTG T GAATTTCACGTACAGATCTGAGTGGTTCTG
desaturase	CAATTTCTTTGTCTAATACTAATAAAGAGA
Nicotiana tabacum	TCTCTTTATTAGTATTAGACAAAGAAATTGCAGAACCACTCAGATCT	2526
Arg36 Term	GTACGTGAAATTC A CAACTTAATGTGAGAGGAGGCAGAAGTGGCT
AGA-TGA	GCCCCAGTTCTGGGCTTAGGGTAGATTCT
	TTAAGTTG T GAATTTCA	2527
	TGAAATTC A CAACTTAA	2528

Reducing linolenic acid	GCGAGTTGGGTTTTATCAGAATGTGGTCTGAGGCCACTCCCGAGG	2529
omega-3 fatty acid	GTCTATCCTAAGCCA T GAACTGGCCACCCTTTGTTGAATTCCAATC
desaturase	CCACAAAGCTGAGATTTTCAAGAACAGATC
Sesamum indicum	GATCTGTTCTTGAAAATCTCAGCTTTGTGGGATTGGAATTCAACAA	2530
Arg22 Term	AGGGTGGCCAGTTC A TGGCTTAGGATAGACCCTCGGGAGTGGCC
AGA-TGA	TCAGACCACATTCTGATAAAACCCAACTCGC
	CTAAGCCA T GAACTGGC	2531
	GCCAGTTC A TGGCTTAG	2532

Reducing linolenic acid	CAGAATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAA	2533
omega-3 fatty acid	GAACTGGCCACCCTT A GTTGAATTCCAATCCCACAAAGCTGAGATT
desaturase	TTCAAGAACAGATCTTGGAAATGGTTCTTC
Sesamum indicum	GAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGTGG	2534
Leu27 Term	GATTGGAATTCAAC T AAGGGTGGCCAGTTCTTGGCTTAGGATAGA
TTG-TAG	CCCTCGGGAGTGGCCTCAGACCACATTCTG
	CCACCCTT A GTTGAATT	2535
	AATTCAAC T AAGGGTGG	2536

Reducing linolenic acid	AATGTGGTCTGAGGCCACTCCCGAGGGTCTATCCTAAGCCAAGAA	2537
omega-3 fatty acid	CTGGCCACCCTTTGT A GAATTCCAATCCCACAAAGCTGAGATTTTC
desaturase	AAGAACAGATCTTGGAAATGGTTCTTCATT
Sesamum indicum	AATGAAGAACCATTTCCAAGATCTGTTCTTGAAAATCTCAGCTTTGT	2538
Leu28 Term	GGGATTGGAATTC T ACAAAGGGTGGCCAGTTCTTGGCTTAGGATA
TTG-TAG	GACCCTCGGGAGTGGCCTCAGACCACATT
	CCCTTTGT A GAATTCCA	2539
	TGGAATTC T ACAAAGGG	2540

Reducing linolenic acid	CTCCCGAGGGTCTATCCTAAGCCAAGAACTGGCCACCCTTTGTTG	2541
omega-3 fatty acid	AATTCCAATCCCACA T AGCTGAGATTTTCAAGAACAGATCTTGGAA
desaturase	ATGGTTCTTCATTCTGTTTGTCGAGTGGGA
Sesamum indicum	TCCCACTCGACAAACAGAATGAAGAACCATTTCCAAGATCTGTTCT	2542
Lys34 Term	TGAAAATCTCAGCT A TGTGGGATTGGAATTCAACAAAGGGTGGCC
AAG-TAG	AGTTCTTGGCTTAGGATAGACCCTCGGGAG
	ATCCCACA T AGCTGAGA	2543
	TCTCAGCT A TGTGGGAT	2544

Reducing linolenic acid	CATCAGAGCGGCGATACCTAAGCATTGCTGGGTTAAGAATCCATG	2545
omega-3 fatty acid	GAAGTCTATGAGTTA G GTCGTCAGAGAGCTAGCCATCGTGTTCGC
desaturase	ACTAGCTGCTGGAGCTGCTTACCTCAACAAT
Brassica napus	ATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACACGATGGC	2546
Tyr3 Term	TAGCTCTCTGACGAC C TAACTCATAGACTTCCATGGATTCTTAACC
TAC-TAG	CAGCAATGCTTAGGTATCGCCGCTCTGATG
	ATGAGTTA G GTCGTCAG	2547
	CTGACGAC C TAACTCAT	2548

Reducing linolenic acid	GCGGCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCT	2549
omega-3 fatty acid	ATGAGTTACGTCGTC T GAGAGCTAGCCATCGTGTTCGCACTAGCT
desaturase	GCTGGAGCTGCTTACCTCAACAATTGGCTTG
Brassica napus	CAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGAACA	2550
Arg6 Term	CGATGGCTAGCTCTC A GACGACGTAACTCATAGACTTCCATGGAT
AGA-TGA	CTTAACCCAGCAATGCTTAGGTATCGCCGC
	ACGTCGTC T GAGAGCTA	2551
	TAGCTCTC A GACGACGT	2552

Reducing linolenic acid	GCGATACCTAAGCATTGCTGGGTTAAGAATCCATGGAAGTCTATGA	2553
omega-3 fatty acid	GTTACGTCGTCAGA T AGCTAGCCATCGTGTTCGCACTAGCTGCTG
desaturase	GAGCTGCTTACCTCAACAATTGGCTTGTTT
Brassica napus	AAACAAGCCAATTGTTGAGGTAAGCAGCTCCAGCAGCTAGTGCGA	2554
Glu7 Term	ACACGATGGCTAGCT A TCTGACGACGTAACTCATAGACTTCCATG
GAG-TAG	GATTCTTAACCCAGCAATGCTTAGGTATCGC
	TCGTCAGA T AGCTAGCC	2555
	GGCTAGCT A TCTGACGA	2556

Reducing linolenic acid	CCATGGAAGTCTATGAGTTACGTCGTCAGAGAGCTAGCCATCGTG	2557
omega-3 fatty acid	TTCGCACTAGCTGCT T GAGCTGCTTACCTCAACAATTGGCTTGTTT
desaturase	GGCCTCTCTATTGGATTGCTCAAGGAACCA
Brassica napus	TGGTTCCTTGAGCAATCCAATAGAGAGGCCAAACAAGCCAATTGTT	2558
Gly17 Term	GAGGTAAGCAGCTC A AGCAGCTAGTGCGAACACGATGGCTAGCT
GGA-TGA	CTCTGACGACGTAACTCATAGACTTCCATGG
	TAGCTGCT T GAGCTGCT	2559
	AGCAGCTC A AGCAGCTA	2560

Reducing linolenic acid	GCAAGTTGGGTTCTATCAGAATGTGGTCTTAGACCACTACCAAGAA	2561
omega-3 fatty acid	TATACCCAAAGCCC T GAATAGGGTCTTCTTCCGTTTGCGCCACCAA
desaturase	TTTAAATCTGAGAAGAATTTCACCTTCAC
Solanum tuberosum	GTGAAGGTGAAATTCTTCTCAGATTTAAATTGGTGGCGCAAACGGA	2562
Arg22 Term	AGAAGACCCTATTC A GGGCTTTGGGTATATTCTTGGTAGTGGTCTA
AGA-TGA	AGACCACATTCTGATAGAACCCAACTTGC
	CAAAGCCC T GAATAGGG	2563
	CCCTATTC A GGGCTTTG	2564

Reducing linolenic acid	TGGTCTTAGACCACTACCAAGAATATACCCAAAGCCCAGAATAGG	2565
omega-3 fatty acid	GTCTTCTTCCGTTTG A GCCACCAATTTAAATCTGAGAAGAATTTCA
desaturase	CCTTCACCTATACGAACAGATCGGAATTGT
Solanum tuberosum	ACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAATTCTTCTCAGA	2566
Cys29 Term	TTTAAATTGGTGGC T CAAACGGAAGAAGACCCTATTCTGGGCTTTG
TGC-TGA	GGTATATTCTTGGTAGTGGTCTAAGACCA
	TCCGTTTG A GCCACCAA	2567
	TTGGTGGC T CAAACGGA	2568

Reducing linolenic acid	CACTACCAAGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGT	2569
omega-3 fatty acid	TTGCGCCACCAATT G AAATCTGAGAAGAATTTCACCTTCACCTATA
desaturase	CGAACAGATCGGAATTGTTGGGCATTGAG
Solanum tuberosum	CTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGAAGGTGAAA	2570
Leu33 Term	TTCTTCTCAGATTT C AATTGGTGGCGCAAACGGAAGAAGACCCTAT
TTA-TGA	TCTGGGTTTGGGTATATTCTTGGTAGTG
	CACCAATT G AAATCTGA	2571
	TCAGATTT C AATTGGTG	2572

Reducing linolenic acid	AGAATATACCCAAAGCCCAGAATAGGGTCTTCTTCCGTTTGCGCCA	2573
omega-3 fatty acid	CCAATTTAAATCTG T GAAGAATTTCACCTTCACCTATACGAACAGAT
desaturase	CGGAATTGTTGGGCATTGAGGGTAAGTG
Solanum tuberosum	CACTTACCCTCAATGCCCAACAATTCCGATCTGTTCGTATAGGTGA	2574
Arg36 Term	AGGTGAAATTCTTC A CAGATTTAAATTGGTGGCGCAAACGGAAGAA
AGA-TGA	GACCCTATTCTGGGCTTTGGGTATATTCT
	TAAATCTG T GAAGAATT	2575
	AATTCTTC A CAGATTTA	2576

Reducing linolenic acid	CTCTTTATTATCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACC	2577
omega-3 fatty acid	TATGGCAAGTTG A GTGATTTCAGAATGTGGGCTAAGGCCACTTCC
desaturase	AAGAATCTATGCCAGGCCCAGAAGTGGA
Petroselinum crispum	TCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGCCTTAGCCC	2578
Trp4 Term	ACATTCTGAAATCAC T CAACTTGCCATAGGTGACTCAGAACTCAAA
TGG-TGA	AAAAACAAAGAAGAGGAGGATAATAAAGAG
	GCAAGTTG A GTGATTTC	2579
	GAAATCAC T CAACTTGC	2580

Reducing linolenic acid	TATCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCA	2581
omega-3 fatty acid	AGTTGGGTGATTT G AGAATGTGGGCTAAGGCCACTTCCAAGAATC
desaturase	TATGCCAGGCCCAGAAGTGGAGCTTCATG
Petroselinum crispum	CATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTGGC	2582
Ser7 Term	CTTAGCCCACATTCT C AAATCACCCAACTTGCCATAGGTGACTCAG
TCA-TGA	AACTCAAAAAAAACAAAGAAGAGGAGGATA
	GGTGATTT G AGAATGTG	2583
	CACATTCT C AAATCACC	2584

Reducing linolenic acid	TCCTCCTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCAAG	2585
omega-3 fatty acid	TTGGGTGATTTCA T AATGTGGGCTAAGGCCACTTCCAAGAATCTAT
desaturase	GCCAGGCCCAGAAGTGGAGCTTCATGTT
Petroselinum crispum	AACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGGAAGTG	2586
Glu8 Term	GCCTTAGCCCACATT A TGAAATCACCCAACTTGCCATAGGTGACTC
GAA-TAA	AGAACTCAAAAAAAACAAAGAAGAGGAGGA
	TGATTTCA T AATGTGGG	2587
	CCCACATT A TGAAATCA	2588

Reducing linolenic acid	CTCTTCTTTGTTTTTTTTGAGTTCTGAGTCACCTATGGCAAGTTGGG	2589
omega-3 fatty acid	TGATTTCAGAAT G AGGGCTAAGGCCACTTCCAAGAATCTATGCCA
desaturase	GGCCCAGAAGTGGAGCTTCATGTTTCAAC
Petroselinum crispum	GTTGAAACATGAAGCTCCACTTCTGGGCCTGGCATAGATTCTTGG	2590
Cys9 Term	AAGTGGCCTTAGCCC T CATTCTGAAATCACCCAACTTGCCATAGGT
TGT-TGA	GACTCAGAACTCAAAAAAAACAAAGAAGAG
	TCAGAATG A GGGCTAAG	2591
	CTTAGCCC T CATTCTGA	2592

Reducing linolenic acid	ATGAAGCAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTA	2593
omega-3 fatty acid	ATGGTTTTCATGCT T AAGAAGAAGAAGAAGAAGAGGATTTCGACTT
desaturase	AAGCAATCCTCCTCCATTCAATATTGGTC
Vernicia fordii	GACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTCTTC	2594
Lys21 Term	TTCTTCTTCTTCTT A AGCATGAAAACCATTAACGCCATTTAGAATTG
AAA-TAA	GGGTGTCTTTGTACTGTTGCTGCTTCAT
	TTCATGCT T AAGAAGAA	2595
	TTCTTCTT A AGCATGAA	2596

Reducing linolenic acid	AAGCAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATG	2597
omega-3 fatty acid	GTTTTCATGCTAAA T AAGAAGAAGAAGAAGAGGATTTCGACTTAAG
desaturase	CAATCCTCCTCCATTCAATATTGGTCAGA
Vernicia fordii	TCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATCCTC	2598
Glu22 Term	TTCTTCTTCTTCTT A TTTAGCATGAAAACCATTAACGCCATTTAGAA
GAA-TAA	TTGGGGTGTCTTTGTACTGTTGCTGCTT
	ATGCTAAA T AAGAAGAA	2599
	TTCTTCTT A TTTAGCAT	2600

Reducing linolenic acid	CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT	2601
omega-3 fatty acid	TTCATGCTAAAGAA T AAGAAGAAGAAGAGGATTTCGACTTAAGCAA
desaturase	TCCTCCTCCATTCAATATTGGTCAGATCC
Vernicia fordii	GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC	2602
Glu23 Term	CTCTTCTTCTTCTT A TTCTTTAGCATGAAAACCATTAACGCCATTTA
GAA-TAA	GAATTGGGGTGTCTTTGTACTGTTGCTG
	CTAAAGAA T AAGAAGAA	2603
	TTCTTCTT A TTCTTTAG	2604

Reducing linolenic acid	CAGCAACAGTACAAAGACACCCCAATTCTAAATGGCGTTAATGGTT	2605
omega-3 fatty acid	TTCATGCTAAAGAA T AAGAAGAAGAAGAGGATTTCGACTTAAGCAA
desaturase	TCCTCCTCCATTCAATATTGGTCAGATCC
Vernicia fordii	GGATCTGACCAATATTGAATGGAGGAGGATTGCTTAAGTCGAAATC	2606
Glu24 Term	CTCTTCTTCTTCTT A TTCTTTAGCATGAAAACCATTAACGCCATTTA
GAA-TAA	GAATTGGGGTGTCTTTGTACTGTTGCTG
	CTAAAGAA T AAGAAGAA	2607
	TTCTTCTT A TTCTTTAG	2608

Reducing linolenic acid	GGTCCAAGCACAGCCTCTACAACATGTTGGTAATGGTGCAGGGAA	2609
omega-3 fatty acid	AGAAGATCAAGCTTA G TTTGATCCAAGTGCTCCACCACCCTTCAAG
desaturase	ATTGCAAATATCAGAGCAGCAATTCCAAAA
Glycine max	TTTTGGAATTGCTGCTCTGATATTTGCAATCTTGAAGGGTGGTGGA	2610
Tyr21 Term	GCACTTGGATCAAA C TAAGCTTGATCTTCTTTCCCTGCACCATTAC
TAT-TAG	CAACATGTTGTAGAGGCTGTGCTTGGACC
	CAAGCTTA G TTTGATCC	2611
	GGATCAAA C TAAGCCTG	2612

Reducing linolenic acid	GGTAATGGTGCAGGGAAAGAAGATCAAGCTTATTTTGATCCAAGT	2613
omega-3 fatty acid	GCTCCACCACCCTTC T AGATTGCAAATATCAGAGCAGCAATTCCAA
desaturase	AACATTGCTGGGAGAAGAACACATTGAGAT
Glycine max	ATCTCAATGTGTTCTTCTCCCAGCAATGTTTTGGAATTGCTGCTCT	2614
Lys31 Term	GATATTTGCAATCT A GAAGGGTGGTGGAGCACTTGGATCAAAATAA
AAG-TAG	GCTTGATCTTCTTTCCCTGCACCATTACC
	CACCCTTC T AGATTGCA	2615
	TGCAATCT A GAAGGGTG	2616

Reducing linolenic acid	AAAGAAGATCAAGCTTATTTTGATCCAAGTGCTCCACCACCCTTCA	2617
omega-3 fatty acid	AGATTGCAAATATC T GAGCAGCAATTCCAAAACATTGCTGGGAGAA
desaturase	GAACACATTGAGATCTCTGAGTTATGTTC
Glycine max	GAACATAACTCAGAGATCTCAATGTGTTCTTCTCCCAGCAATGTTTT	2618
Arg36 Term	GGAATTGCTGCTC A GATATTTGCAATCTTGAAGGGTGGTGGAGCA
AGA-TGA	CTTGGATCAAAATAAGCTTGATCTTCTTT
	CAAATATC T GAGCAGCA	2619
	TGCTGCTC A GATATTTG	2620

Reducing linolenic acid	TATTTTGATCCAAGTGCTCCACCACCCTTCAAGATTGCAAATATCA	2621
omega-3 fatty acid	GAGCAGCAATTCCA T AACATTGCTGGGAGAAGAACACATTGAGAT
desaturase	CTCTGAGTTATGTTCTGAGGGATGTGTTGG
Glycine max	CCAACACATCCCTCAGAACATAACTCAGAGATCTCAATGTGTTCTT	2622
Leu41 Term	CTCCCAGCAATGTT A TGGAATTGCTGCTCTGATATTTGCAATCTTG
AAA-TAA	AAGGGTGGTGGAGCACTTGGATCAAAATA
	CAATTCCA T AACATTGC	2623
	GCAATGTT A TGGAATTG	2624

Reducing linolenic acid	CATCCACCCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGC	2625
omega-3 fatty acid	CCGGCTCGTGCTCTCC T AGTGCTCGGGCCTCGCGCCCGTCCGCC
desaturase	GCCTGCGCGCCGGCCGGGGCGCCATTGCGGCGC
Zea mays	GCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGGACGG	2626
Glu8 Term	GCGCGAGGCCCGAGCACT A GGAGAGCACGAGCCGGGCCATTGC
GAG-TAG	CGCCGTCAGCGGGGCGGGTGCGGGTGCGGGTGGATG
	TGCTCTCC T AGTGCTCG	2627
	CGAGCACT A GGAGAGCA	2628

Reducing linolenic acid	ACCCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGG	2629
omega-3 fatty acid	CTCGTGCTCTCCGAGTG A TCGGGCCTCGCGCCCGTCCGCCGCCT
desaturase	GCGCGCCGGCCGGGGCGCCATTGCGGCGCGGTCA
Zea mays	TGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGCGG	2630
Cys9 Term	ACGGGCGCGAGGCCCGA T CACTCGGAGAGCACGAGCCGGGCCA
TGC-TGA	TTGCCGCCGTCAGCGGGGCGGGTGCGGGTGCGGGT
	TCCGAGTG A TCGGGCCT	2631
	AGGCCCGA T CACTCGGA	2632

Reducing linolenic acid	CCGCACCCGCACCCGCCCCGCTGACGGCGGCAATGGCCCGGCT	2633
omega-3 fatty acid	CGTGCTCTCCGAGTGCT A GGGCCTCGCGCCCGTCCGCCGCCTGC
desaturase	GCGCCGGCCGGGGCGCCATTGCGGCGCGGTCACC
Zea mays	GGTGACCGCGCCGCAATGGCGCCCCGGCCGGCGCGCAGGCGGC	2634
Ser10 Term	GGACGGGCGCGAGGCCC T AGCACTCGGAGAGCACGAGCCGGGC
TCG-TAG	CATTGCCGCCGTCAGCGGGGCGGGTGCGGGTGCGG
	CGAGTGCT A GGGCCTCG	2635
	CGAGGCCC T AGCACTCG	2636

Reducing linolenic acid	GCTCGGGCCTCGCGCCCGTCCGCCGCCTGCGCGCCGGCCGGGG	2637
omega-3 fatty acid	CGCCATTGCGGCGCGGT G ACCCCCCGCGCTCTCCGCGGCGCCG
desaturase	CGCCGTCGTCCCGCGTCCGCGTCCATCCACCGCGA
Zea mays	TCGCGGTGGATGGACGCGGACGCGGGACGACGGCGCGGCGCCG	2638
Ser29 Term	CGGAGAGCGCGGGGGGT C ACCGCGCCGCAATGGCGCCCCGGCC
TCA-TGA	GGCGCGCAGGCGGCGGACGGGCGCGAGGCCCGAGC
	GGCGCGGT G ACCCCCCG	2639
	CGGGGGGT C ACCGCGCC	2640

Reducing linolenic acid	CCCCCTCCCCCACGCACACGCACAGATCCATCCGCGGCCATGGC	2641
omega-3 fatty acid	CCCCGCAATGAGGCCG T AGCAGGAGGCGAGCTGCAAGGCCACCG
desaturase	AGGACCACCGCTCCGAGTTCGACGCCGCCAAGC
Triticum aestivum	GCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGGCCTTG	2642
Glu8 Term	CAGCTCGCCTCCTGCT A CGGCCTCATTGCGGGGGCCATGGCCGC
GAG-TAG	GGATGGATCTGTGCGTGTGCGTGGGGGAGGGGG
	TGAGGCCG T AGCAGGAG	2643
	CTCCTGCT A CGGCCTCA	2644

Reducing linolenic acid	CCTCCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCC	2645
omega-3 fatty acid	CGCAATGAGGCCGGAG T AGGAGGCGAGCTGCAAGGCCACCGAG
desaturase	GACCACCGCTCCGAGTTCGACGCCGCCAAGCCGC
Triticum aestivum	GCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGTGGCC	2646
Gln9 Term	TTGCAGCTCGCCTCCT A CTCCGGCCTCATTGCGGGGGCCATGGC
CAG-TAG	CGCGGATGGATCTGTGCGTGTGCGTGGGGGAGG
	GGCCGGAG T AGGAGGCG	2647
	CGCCTCCT A CTCCGGCC	2648

Reducing linolenic acid	CCCCCACGCACACGCACAGATCCATCCGCGGCCATGGCCCCCGC	2649
omega-3 fatty acid	AATGAGGCCGGAGCAG T AGGCGAGCTGCAAGGCCACCGAGGACC
desaturase	ACCGCTCCGAGTTCGACGCCGCCAAGCCGCCGC
Triticum aestivum	GCGGCGGCTTGGCGGCGTCGAACTCGGAGCGGTGGTCCTCGGT	2650
Glu10 Term	GGCCTTGCAGCTCGCCT A CTGCTCCGGCCTCATTGCGGGGGCCA
GAG-TAG	TGGCCGCGGATGGATCTGTGCGTGTGCGTGGGGG
	CGGAGCAG T AGGCGAGC	2651
	GCTCGCCT A CTGCTCCG	2652

Reducing linolenic acid	ACGCACAGATCCATCCGCGGCCATGGCCCCCGCAATGAGGCCGG	2653
omega-3 fatty acid	AGCAGGAGGCGAGCTG A AAGGCCACCGAGGACCACCGCTCCGA
desaturase	GTTCGACGCCGCCAAGCCGCCGCCCTTCCGCATC
Triticum aestivum	GATGCGGAAGGGCGGCGGCTTGGCGGCGTCGAACTCGGAGCGG	2654
Cys13 TermTGGTCCTCGGTGGCCTT T CAGCTCGCCTCCTGCTCCGGCCTCATT
TGC-TGA	GCGGGGGCCATGGCCGCGGATGGATCTGTGCGT
	GCGAGCTG A AAGGCCAC	2655
	GTGGCCTT T CAGCTCGC	2656

Reducing linolenic acid	CTTCACAAATCACAAATCGGAATCAGATCCACCACGACACCCCGG	2657
omega-3 fatty acid	CGGCAATGGCGGCGT A GGCGACCCAGGAGGCCGACTGCAAGGC
desaturase	TTCCGAGGACGCCCGTCTCTTCTTCGACGCCGC
Oryza sativa	GCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGCCTTGCAGTC	2658
Ser4 Term	GGCCTCCTGGGTCGCC T ACGCCGCCATTGCCGCCGGGGTGTCGT
TCG-TAG	GGTGGATCTGATTCCGATTTGTGATTTGTGAAG
	GGCGGCGT A GGCGACCC	2659
	GGGTCGCC T ACGCCGCC	2660

Reducing linolenic acid	ATCACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATG	2661
omega-3 fatty acid	GCGGCGTCGGCGACC T AGGAGGCCGACTGCAAGGCTTCCGAGGA
desaturase	CGCCCGTCTCTTCTTCGACGCCGCCAAGCCCC
Oryza sativa	GGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGAAGC	2662
Gln7 Term	CTTGCAGTCGGCCTCCT A GGTCGCCGACGCCGCCATTGCCGCCG
CAG-TAG	GGGTGTCGTGGTGGATCTGATTCCGATTTGTGAT
	CGGCGACC T AGGAGGCC	2663
	GGCCTCCT A GGTCGCCG	2664

Reducing linolenic acid	ACAAATCGGAATCAGATCCACCACGACACCCCGGCGGCAATGGC	2665
omega-3 fatty acid	GGCGTCGGCGACCCAG T AGGCCGACTGCAAGGCTTCCGAGGACG
desaturase	CCCGTCTCTTCTTCGACGCCGCCAAGCCCCCGC
Oryza sativa	GCGGGGGCTTGGCGGCGTCGAAGAAGAGACGGGCGTCCTCGGA	2666
Glu8 Term	AGCCTTGCAGTCGGCCT A CTGGGTCGCCGACGCCGCCATTGCCG
GAG-TAG	CCGGGGTGTCGTGGTGGATCTGATTCCGATTTGT
	CGACCCAG T AGGCCGAC	2667
	GTCGGCCT A CTGGGTCG	2668

Reducing linolenic acid	TCAGATCCACCACGACACCCCGGCGGCAATGGCGGCGTCGGCGA	2669
omega-3 fatty acid	CCCAGGAGGCCGACTG A AAGGCTTCCGAGGACGCCCGTCTCTTC
desaturase	TTCGACGCCGCCAAGCCCCCGCCCTTCCGCATC
Oryza sativa	GATGCGGAAGGGCGGGGGCTTGGCGGCGTCGAAGAAGAGACGG	2670
Cys10 Term	GCGTCCTCGGAAGCCTT T CAGTCGGCCTCCTGGGTCGCCGACGC
TGC-TGA	CGCCATTGCCGCCGGGGTGTCGTGGTGGATCTGA
	GCCGACTG A AAGGCTTC	2671

Claims

What is claimed is:

1. An oligonucleotide for targeted alteration of genetic sequence, comprising a single-stranded oligonucleotide having a DNA domain, said DNA domain having at least one mismatch with respect to the genetic sequence to be altered, and further comprising chemical modifications of the oligonucleotide, said chemical modifications selected from the group consisting of an o-methyl modification, an LNA modification including LNA derivatives and analogs, two or more phosphorothioate linkages on a terminus, and a combination of any two or more of these modifications.

2. The oligonucleotide according to claim one that comprises two or more phosphorothioate linkages on at least the 3′ terminus.

3. The oligonucleotide according to claim one that comprises a 2′-O-methyl analog.

4. The oligonucleotide according to claim one that comprises an LNA nucleotide, including an LNA derivative or analog.

5. The oligonucleotide according to claim one that comprises a combination of at least two modifications selected from the group of a phosphorothioate linkage, a 2′-O-methyl analog, a locked nucleotide analog and a ribonucleotide.

6. The oligonucleotide according to any one of claims 1 to 5 that comprises at least one unmodified ribonucleotide.

7. The oligonucleotide according to any one of claims 1 to 6, wherein the sequence of said oligonucleotide is selected from the group consisting of SEQ ID NOS: 1-2672.

8. A method of targeted alteration of genetic material, comprising combining the target genetic material with an oligonucleotide according to any one of claims 1 to 7 in the presence of purified proteins.

9. A method of targeted alteration of genetic material, comprising administering to a cell extract an oligonucleotide of any one of claims 1 to 7.

10. A method of targeted alteration of genetic material, comprising administering to a cell an oligonucleotide of any one of claims 1 to 7.

11. A method of targeted alteration of genetic sequence in callus, comprising administering to the callus an oligonucleotide of any one of claims 1 to 7.

12. A method of targeted alteration of genetic sequence, comprising combining target genetic material with an oligonucleotide according to any one of claims 1 to 7, said target genetic material being a non-transcribed DNA strand of a duplex DNA.

13. The genetic material obtained by any one of the methods of claim 8, 9 or claim 10.

14. A cell comprising the genetic material of claim 13.

15. A plant organism comprising the cell according to claim 14.

16. A plant or plant part produced by the method of claim 11.

17. A method of determining whether an oligonucleotide is optimized for targeted alteration of a genetic sequence, which comprises:

(a) comparing the efficiency of alteration of a targeted genetic sequence by an oligonucleotide of any one of claims 1 to 7 with the efficiency of alteration of the same targeted genetic sequence by a second oligonucleotide, said second oligonucleotide selected from the group of an oligonucleotide that lacks the mismatch, a fully modified phosphorothiolated oligonucleotide, a fully modified 2′-O-methylated oligonucleotide and a chimeric double-stranded double hairpin containing RNA and DNA nucleotides.

18. The method of claim 17 in which the alteration is produced in a plant cell extract.

19. The method of claim 17 in which the alteration is produced in a cell.

20. A kit comprising the oligonucleotide according to any one of claims 1 to 7 and a second oligonucleotide selected from the group of an oligonucleotide that lacks the mismatch, a fully modified phosphorothiolated oligonucleotide, a fully modified 2-O-methylated oligonucleotide and a chimeric double stranded double hairpin containing RNA and DNA nucleotides.