MXPA01001224A - Plants, seeds and oils having an elevated total monounsaturated fatty acid content - Google Patents

Abstract

Plants, seeds and oils having a total long-chain monounsaturated content of at least about 82%and an erucic acid content of at least about 15%are described. Methods for producing plants having the profiled fatty acid content are also described.

Description

PLANTS, SEEDS AND OILS WHICH HAVE A TOTAL, ELEVATED MONOINSATURATED FATTY ACID CONTENT Field of the Invention This invention relates to fatty acid desaturases and desaturase proteins that encode nucleic acids. More particularly, the invention relates to the fatty acid delta-12 and delta-15 fatty acid proteins encoding said nucleic acid affecting the fatty acid composition in plants, polypeptides produced from said nucleic acids and plants expressing said nucleic acids. nucleic acids.

BACKGROUND OF THE INVENTION Many cultivation studies have been conducted to improve the fatty acid profile of the Brassica varieties. Pleines and Freidt, in Faí Sci. Technol. , 90 (5), pages 1 67 to 171 (1 988) reported plañías lines with reduced levels of C? 8: 3 (2.5-5.8%) combined with a high oleic content (73 to 79%). Rakow and McGregor, in J. Amer. Oil Chem. Soc, 50, pages 400 to 403 (October 1973) discuss the problems associated with the mulenols that select linoleic and linolenic acids. In Can. J. Plant Sci., 68, pages 509 to 51 1 (April 1988) describes summer stellar rapeseed that produces seed oil with 3% linolenic acid and 28% linoleic acid. Roy and Tarr, in Z. Pflanzenzuchtg, 95 (3), pages 201 to 209 (1 985) report the transfer of genes through an interspecific crossing of Brassica júncea in Brassica napus that results in a reconstituted line that combines a content high linoleic acid with a low linolenic acid condenide. Roy and Tarr, in Plañí Breeding, 98, pages 89 to 96 (1 987) explain the prospects for the development of B. napus L. which has an improved linolenic content and linolenic acid. The European patent application 323,753 published on July 12, 1989 discloses oil seeds that are more than 79% oleic acid combined with less than 3.5% linolenic acid. Canvin, in Can. J. Boíany, 43, pages 63 to 69 (1965) explains the effect of temperature on the fatty acid composition of oils from crops of several seeds including rapeseed. The mulations are generally induced with an extremely high dose of radiation and / or chemical mutagenesis (Gaul, H. Radiation Botany (1964) 4: pages 155 to 232). High dose levels, which exceed the LD5o. and generally reach LD90, they lead to the maximum mulation rates that can be achieved. In the culture of the Brassica mutation, high levels of chemical mutagenes alone or in combination with radiation have induced a limited number of fatty acid mutations (Rakow, G.Z. Pflanzenzuchfg (1973) 69: pages 62 to 82). The low a-linolenic acid mutation derived from the Rakow mutation culture program did not have a direct commercial application due to the low seed production. The first commercial crop using the low a-linolenic acid mutation derived in 1973 was released in 1988 as the Stelar variety (Scarth, R. et al, Can. J. Plant Sci. (1988) 68: pages 509 to 51 1 ). The Esíelar crop was 20% less than the commercial cullives at the time of its release. Alterations in the fatty acid composition of vegetable oils are desirable to meet specific food and industrial uses. For example, varieties of Brassica cañola with increased monounsaturated levels (oleic acid) in the seed oil, and the products derived from said oil, would improve the lipid content. Canola lines, which are low in polyunsaturated fatty acids and high in oleic acid, tend to have a higher oxidative stability, which is a useful characteristic for the retail food industry. The useful characteristics of vegetable oils for industrial uses as lubrication fluids include a desirable behavior at low temperature such as a low point of spillage, and a low point of nebulosity together with a very high oxidative stability. The delta-12 fatty acid desaturase (also known as oleic desaturase) is comprised in the enzymatic conversion of oleic acid to linoleic acid. The delta-15 fatty acid desalurase (also known as linoleic acid desaturase) is comprised in the enzymatic conversion of linoleic acid to a-linolenic acid. The delta-12 microsomal desaturase has been cloned and characterized using T-DNA labeling. Okuley, et al, in Plant Cell 6: pages 147 to 158 (1994). In WO 94/1 1516 of Lighier et al, the nucleotide sequences of the highest genes of the plants coding for the delta-1 2 fatty acid microsomal desaturase are described. The sequences of the highest genes of the plants which encode the microsomal fatty acid desaturases and plastids delta-15 are described in N. Yadav, et al, in Plant Physiol., 103: pages 467 to 476 (1 993), WO 93/1 1245 and by V. Arondel, et al, in Science, 258: pages 1353 to 1355 (1992).

Summary of the Invention Iriacylglycerols containing fatty acids with heterogeneous chain lengths and monounsaturated ally levels can provide useful characteristics for industrial purposes. Plants with fatty acid compositions having high monounsaturated levels, and heterogeneous lengths of the chains would provide a source of industrial oils for uses such as lubrication. In one aspect, the invention features a Brassica plant, and offspring thereof, which produces seeds having a monounsaturated long chain fatty acid content of at least about 82% and an erucic acid content of at least about approximately 15% based on the loyal composition of fatty acid. The content of oleic acid and eicosoic acid of the seeds is at least about 37% and at least about 14%, based on the total fatty acid composition, respectively. The saturated fatty acid content of said seeds is less than 7% and the content of polyunsaturated fatty acid is less than about 1%. In some embodiments, the plants have a monounsaturated fatty acid content of about 85% to about 90%, and an erucic acid content of at least about 15% based on the total fatty acid composition. In such plants, the content of oleic acid can be at least about 42% and in particular, about 47% to about 56% based on the total fatty acid composition. The erucic acid content is from about 17% to about 31%, and the content of eicosenoic acid is about 15% up to about 21%. The invention also characterizes a Brassica seed oil having a long chain monounsaturated fatty acid conent of at least about 82% and an erucic acid content of at least about 15% based on the total acid composition fatty. Such oils may have an oleic acid and eicosoic acid content of at least about 14% and 37%, respectively, based on the total fatty acid composition. The saturated fatty acid content may be less than about 7%, for example, less than about 4% or about 2 to 4%. The content of polyunsaturated fatty acid is less than about 11% and in particular embodiments, less than 9%, based on the total fatty acid composition. The α-linolenic acid content may be from about 1% to about 2%. In some embodiments, Brassica seed oil contains a long chain monounsaturated fatty acid content of about 85% to about 90%. In such oils, the content of oleic acid is at least about 42% and in particular embodiments, it is from about 47% to about 56%, based on the total composition of the fatty acid. The content of erucic acid and eicosenoic acid is approximately 17% > up to about 31% and from about 15% to about 21%, respectively, based on the total fatty acid composition. A Brassica seed oil having a long chain monounsaturated fatty acid content of at least about 82% has also been characterized, wherein the sum of the content of nervonic acid, erucic acid and eicosenoic acid is approximately 50%. % to approximately 66% based on the total fatty acid composition. Said seed oil may have an oleic acid content of about 25% up to about 30%. The present invention also features a method of producing plants that have a long-chain monounsaturated fatty acid condense of at least about 82% and an erucic acid content of at least about 15%, based on the total fatty acid composition. Methods include crossing a first line of plants with a second line of plants and selecting the offspring with the desired composition of fatty acid. The first plant line has an erucic acid content of at least about 45%. The second plant line has an oleic acid content of at least about 84%. The invention also features a method for making a vegetable oil. The method comprises the steps of grinding Brassica seeds having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 1 5% based on the total composition of fatty acid, and the extraction of vegetable oil from ground seeds. The method can also include the oil refining and blanching steps, and oil deodorization. The invention also features a lubricating or hydraulic fluid comprising a Brassica oil having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% based on the total fatty acid composition, and an additive. The additive may be an antioxidant, a rust inhibitor, a corrosion inhibitor, a pour point depressant, an anti-foam additive, a colorant and a detergent. The additive may be present in an amount of about 0.01% to about 20% by weight.

Brief Description of the Sequence Listing The SEQUENCE IDENTIFICATION NO: 1 illustrates the DNA sequence for coding the region of a wild-type Brassica Fad2-D gene. SEQUENCE IDENTIFICATION NO: 2 is the amino acid sequence deduced for SEQUENCE IDENTIFICATION NO: 1. The SEQUENCE IDENTIFICATION NO: 3 illustrates the DNA sequence for the coding of the mutant region I MC 129 of the Brassica Fad2-D gene. The SEQUENCE IDENTIFICATION NO: 4 is the amino acid sequence deduced for the SEQUENCE IDENTIFICATION NO: 3. SEQUENCE IDENTIFICATION NO: 5 illustrates the sequence of DNA for the coding of the region of a wild-type Brassica Fad2-F gene. SEQUENCE IDENTIFICATION NO: 6 is the amino acid sequence deduced for SEQUENCE IDENTIFICATION NO: 5. SEQUENCE IDENTIFICATION NO: 7 illustrates the DNA sequence for the coding of the Q508 mutant region of the Brassica Fad2-F gene. SEQUENCE IDENTIFICATION NO: 8 is the amino acid sequence deduced for SEQUENCE IDENTIFICATION NO: 7. SEQUENCE IDENTIFICATION NO: 9 illustrates the DNA sequence for the coding of the Q4275 mutant region of the Brassica Fad2-F gene. SEQUENCE IDENTIFICATION NO: 10 is the amino acid sequence deduced for SEQUENCE IDENTIFICATION NO: 9.

Brief Description of the Drawings Figure 1 is a histogram illustrating the frequency distribution of the oleic acid content (C18:.) Of the seed oil in a population that segregates a Weslar junction of Q508 X. The bar marked with WSGA 1 A represents the C18: 1 content of the origin Westar. The bar marked with Q508 represents the content C18:. of the Q508 of origin. Figure 2 illustrates the nucleotide sequences for a Brassica Fad2-D wild-type lipo gene (Fad2-D wt), the mulenle gene IMC129 (Fad2-D GA316 I MC129), the Fad2- gene Wild type F (Fad2-F wt), mulenle gene Q508 (Fad2-F TA51 5 Q508) and mutant Q4275 gene (Fad2-F GA908 Q4275). Figure 3 illustrates the deduced amino acid sequences for the polynucleotides of Figure 2. Figure 4 is a schematic of the culture method used to produce the Brassica pineapples which have a high content of erucic acid and oleic acid.

Detailed Description of the Invention All percentages of fatty acid mentioned herein are percentages by weight of the oil of which the fatty acid is a component. As used in this invention, a "line" is a group of plants that shows little or no genetic variation among individuals of at least one characteristic. These lines can be created by several generations of self-pollination and selection, or vegetative propagation from a single parent using nests or cell culture techniques. As used in this invention, the term "variety" refers to a line which is used for commercial production. The term "mutagenesis" refers to the use of a mutagenic agent to induce random genetic mutations in a population of individuals. The treated population, or a subsequent generation of that population, is then selected by the characteristic (s) that can be used that are the result of the mutations. A "population" is a group of individuals that share a common genetic set. As used in this invention, "M0" is an untraced seed. The term "M 1 is the seed (and the resulting plants) exposed to a mutagenic agent, while" M2"is the offspring (seeds and plants) of self-pollinating My plants," M3"is the offspring of the M2 plants self-polled, and "M4" is the offspring of the self-pollinated M3 plants. "M5" is the offspring of the self-pollinated M4 plants. "M6", "M7", etc. are each the descendants of plañías The term "self-pollination" as used in this invention means "self-pollinating." "Stability" or "stable" as used in this invention means that with respect to a certain fatty acid component., the component is maintained from generation to generation for at least two generations and preferably at least three generations and substantially the same level, for example, preferably ± 5%. The method of the invention has the ability to create lines with improved fatty acid compositions up to 5% generation to generation. The above esiability can be affected by the temperature, location, tension and time of cultivation. Therefore, the comparison of fatty acid profiles should be made from seeds produced under similar growth conditions. Stability can be measured based on the knowledge of the previous generation. Intensive cultivation has produced close Brassica plants whose seed oil contains less than 2% erucic acid. The same varieties have also been grown so that the defatted food contains less than 30 μmol of glucosinolates / grams. "Cañola" as used in this invention, refers to plant seeds or oils which contain less than 2% erucic acid (C22: 1), and results in a defatted food with less than 30 μmol of glucosinolates / grams. The applicants have discovered plañís with mulacíons in a delta-12 fatty acid desaturase gene. Said plans are useful in the fatty acid compositions of the seed oil. Such mutations confer, for example, a high content of oleic acid, and a stabilized, decreased content of lílenic acid, or a high content of oleic acid and a decreased, stabilized content of linolenic acid.

Applicants have additionally discovered plants with mutations in the delia-15 fatty acid desaturase gene. Said plants have useful alterations in the fatty acid composition of the seed oil, for example, a decreased, stabilized level of a-linolenic acid. Applicants have additionally discovered isolated fragments of nucleic acid (polynucleotides) which comprise sequences carrying mutations within the coding sequence of fatty acid desaturases delta-12 or delta-1 5. Mutations confer desirable alterations in acid levels fat in the seed oils of plants that carry such mutations. Delta-12 fatty acid desalurase is also known as omega-6 fatty acid desaturase and we sometimes refer to it as Fad2 or 12-DES. The fatty acid desaturase delta-15 is also known as omega-3 fatty acid desaturase and we refer to it sometimes as Fad3 or 15-DES. A nucleic acid fragment of the invention may be in the form of RNA or in the form of DNA, including cDNA, synthetic DNA or genomic DNA. The DNA can be of two threads or of a single thread, and if it is of a single thread, it can be either an encoder thread or a non-coding thread. An RNA analog may be, for example, mRNA or a combination of ribo- and deoxyribonucleotides. Illustrative examples of a nucleic acid fragment of the invention are the mutani sequences that are illustrated in Figure 3. A fragment of nucleic acid of the invention causes a mutation in a mitochondrial delta-12 fatty acid desaturase encoding the sequence or a mutation in a delta-1 fatty acid microsomal desaturase coding sequence 5. Said mutation produces the resulting non-functional desaturase gene product in the plants, in relation to the function of the gene product encoded by the wild-type sequence. The non-functionality of the delta-12 desaturase gene product can be deduced by the decreased level of the reaction product (linoleic acid) and the increased level of the subsyte (oleic acid) in the tissues of the plant expressing the mutant sequence, compared with corresponding levels in plant tissues expressing the wild-type sequence. The non-functionality of the delta-15 desalurase gene product can be deduced by the decreased level of the reaction product (α-linolenic acid) and the increased level of the subtracter (linoleic acid) in the tissues of the plant expressing the mutant sequence. , compared with the corresponding levels in the plains that express the sequence of the syllable. A nucleic acid fragment of the invention may comprise a portion of the coding sequence, for example, at least about 10 nucleotides, provided that the fragment contains at least one mulation in the coding sequence. The length of a desired fragment depends on the purpose for which the fragment will be used, for example, a PCR preparer, site-driven mutagenesis, and the like. In one embodiment, a nucleic acid fragment of the invention comprises a full length coding sequence of a delian-12 mutant or mutant delta-15 fatty acid desirase, for example, the mutant sequences of Figure 3. In another embodiment , the nucleic acid fragment is from about 20 up to about 50 nucleotides (or base pairs, bp), or from about 50 to about 500 nucleotides, or from about 500 up to about 1200 nucleotides in length. Desirable alterations in the fatty acid levels of the seed oil of plants can be produced using a ribozyme. Ribozyme molecules designed to wash the mRNA transcripts of the delta-1 2 or delta-1 5 desaturase can be used to prevent the expression of delta-12 or delta-15 desaturases. Although several ribozymes can be used that wash the mRNA in the specific recognition sequences at the site to destroy the desaturase of the mRNAs, hammerhead ribozymes are particularly useful. Hammerhead ribozymes clean the mRNAs in places dictated by flanking regions that form the basic complementary pairs with the target mRNA. The only requirement is that the target RNA contains a 5'-UG-3 'nucleotide sequence. The construction and production of hammerhead ribozymes is well known in the art. See, for example, U.S. Pat. No. 5,254,678. Hammerhead ribozyme sequences can be embedded in a stable RNA lal as a transfer RNA (tRNA) to increase washing efficiency in vivo. Consult the study by R. Perriman, et al, in Proc. Nati Acad. Sci. USA, 92 (13): pages 61 75 to 6179 (1,995); the study by R. Feyter, and J. Gaudron, in Methods in Molecular Biology, Vol. 74, Chapter 43, "Expression of Ribozymes in Plañas" (Expressing Ribozymes in Plañís), Edited by Turner, PC, Humana Press Inc., Totowa, NJ. Endoribonucleases such as occur naturally in Tetrahymena thermophila, and which has been described extensively by Cech et al., Are also useful. See, for example, the Patenle U.S. No. 4,987,071. A mutation in a nucleic acid fragment of the invention can be found in any portion of the coding sequence that produces the resulting non-functional gene product. Suitable types of mutations include, without limitation, nucleolide inserts, nucleotide deletions, or transitions and transversions in the wild type coding sequence. Said mutions result in insertions of one or more amino acids, deletions of one or more amino acids, and substitutions of non-conservative amino acids in the corresponding gene product. In some embodiments, the sequence of a nucleic acid fragment may comprise more than one mutation or more than one type of mutation. The introduction or removal of amino acids in a coding sequence can, for example, interrupt the conformation of the alpha-helical or beta-folded essential leaf region of the resulting genetic product. The inlroductions or eliminations of amino acids can also interrupt the bond or catalytic sites important for the activity of the gene product. It is known in the art that the introduction or removal of a larger amount of contiguous amino acids is more likely to produce a non-functional genetic product, compared to a smaller number of amino acids introduced or eliminated. Non-conservative amino acid substitutions can replace an amino acid of one class with an amino acid of a different class. Non-conservative substitutions can make a substantial change in the load or hydrophobicity of the gene product. Non-conservative amino acid substitutions can also make a substantial change in the volume of the side residue chain, for example, by substituting an alanyl residue for an isoleucyl residue. Examples of non-conservative substitutions include substilution of a basic amino acid by a non-polar amino acid, or a polar amino acid by an acidic amino acid. As there are only 20 amino acids encoded in a gene, the substitutions that result in a product of a non-functional gene can be determined by routine experiments, incorporating amino acids of different class in the region of the product of the gene that is targeted for the mutation. Preferred mutations are in a region of the nucleic acid encoding an amino acid sequence motif that is conserved between the delia-12 fatty acid desaturases or the delta-15 fatty acid desirases, such as a His-Xaa-Xaa-motif. Xaa-His (Tables 1 to 3). An example of a suitable region has a conserved HECGH motif that is found, for example, in nucleotides corresponding to amino acids 105 to 09 of the Arabidopsis and delta-12 Brassica desaturase sequences, in the nucleotides corresponding to the amino acids from 101 to 105 of the soybean delta-12 desaturase sequence and in the nucleotides corresponding to the amino acids of 1 1 1 to 1 1 5 of the corn delta-12 desaturase sequence. See, for example, WO 94/1 151 1 6; Okuley et al, in Plant Cell 6: pages 147 to 1 58 (1994). The amino acid designations of a leiras used in this invention are described in the publication by B. Alberts et al, Cell Molecular Biology, 3a. Ed., Garland Publishing, New York, 1994. The amino acids flanking this motif are also highly conserved among delta-12 and delta-15 desaturases and are also suitable candidates for mutations in fragments of the invention. An illustrative modality of a mutation in a nucleic acid fragment of this invention, is a subsumption of Glu by Lis in the HECGH motif of a Brassica delta-12 microsomal desaturase sequence, either in the D form or in the F form. This mutation results in the HJ? CGH sequence being changed. by HKCGH as can be seen by comparing the IDENTIFICATION OF SEQUENCE NO: 2 (form D of wild type) to SEQUENCE IDENTIFICATION NO: 4 (form D mutanfe). A similar mutation is contemplated in other Fad-2 sequences to result in a non-functional gene prodrug. A similar motif can be found in the amino acids 101 to 1 05 of the microsomal desaturase Arabidopsis of the delta-15 fatty acid, as well as in the desaturases of rapeseed and corresponding soybeans (Table 5). See, for example, WO 93/1 1245; the study by V. Arondel et al, in Science, 258: pages 1 153 to 1 1 55 (1992); the study by N. Yadav et al, in Plant Physiol., 103: pages 467 to 476 (1993). Plastid delta-15 fatty acids have a similar motif (Table 5). Among the types of mutations in a HECGH motif that produce the nonfunctional gene product are the non-conservative subsystitutions. An illustrative example of a non-conservative substitution is the substitution of a glycine residue, either by the first or second histidine. This substitution replaces a charged residue (histidine) with a non-polar residue (glycine). Another type of mutation that produces the resulting non-functional gene product is an insert mutation, for example, the insert of a glycine between the cysteine and glutamic acid residues in the HECGH motif. Other regions having suitable molivos of conserved amino acids include the HRRHH motif illustrated in Table 2, the HRTHH motif illustrated in Table 6, and the HVAHH motif illustrated in Table 3. See, for example, WO 94/1 151 16; the study by W. Hiíz el al, in Plañí Physiol., 105: pages 635 to 641 (1994); the study by J. Okuley et al, above; and the study by N. Yadav et al, above. An illustrative example of a mutation in the region shown in Table 3 is a mutation in the nucleotides corresponding to the glycine codon (amino acid 303 of B. napus). A non-conservative substilution from GM to Glu results in an amino acid sequence DRDYG ILNKV being changed by the sequence DRDYEILNKV (compare SEQUENCE IDENTIFICATION NO: 6 form F from wild type to SEQUENCE IDENTIFICATION NO: 10 mutant Q4275, of Figure 3). Another region suitable for a mutation in the sequence of a della-12 desaturase contains the KYLNNP motif in the nucleotides corresponding to the amino acids from 1 71 to 175 of the Brassica desaturase sequence. An illustrative example of a mutation in this region is a substilution of Leu by His, which results in the amino acid sequence (Table 4) KYHNN (compare SEQUENCE IDENTIFICATION NO: 6 Wild-type Fad2-F with SEQUENCE IDENTIFICATION NO: 8 mulante). It is contemplated that a similar mutation in other Fad-2 amino acid sequences results in a non-functional gene product.

TABLE 1 Alignment of Amino Acid Sequences of Fatty Acid Microsomal Desaturases Delta-12 Species Position Amino Acid Sequence Arabidopsis thaliana 100-129 IWVIAHECGH HAFSDYQWLD DTVGLIFHSF Glycine max 96-125 VWVIAHECGH HAFSKYQWVD DWGLTLHST Zea mays 106-135 VWVIAHECGH HAFSDYSLLD DWGLVLHSS Ricinus communis3 1-29 WVMAHDCGH HAFSDYQLLD DWGLILHSC Brassica napus D 100-128 VWVIAHECGH HAFSDYQWLD DTVGLIFHS Brassica napus F 100-128 VWVIAHECGH HAFSDYQWLD DTVGLIFHS of pRF2-1C plasmid TABLE 2 Alignment of Amino Acid Sequences of Microsomal Desaturases of Fatty Acid Delta-12 Species Position Sequence of Amino Acid Arabidopsis thaliana 130-158 LLVPYFSWKY SHRRHHSNTG SLERDEVFV Glycine max 126-154 LLVPYFSWKI SHRRHHSNTG SLDRDEVFV Zea mays SHRRHHSNTG SLERDEVFV Ricinus 136-164 LMVPYFSWKY LLVPYFSWKH SHRRHHSNTG SLERDEVFV communis3 30-58 Brassica napus D 130-158 SHRRHHSNTG SLERDEVFV Brassica napus LLVPYFSWKY F SHRRHHSNTG SLERDEVFV LLVPYFSWKY 130-158 of plasmid pRF2-1C TABLE 3 Alignment of Amino Acid Sequences of microsomal Desalurasas Delta-1 Fatty Acid 2 Species Position Amino Acid Sequence Arabidopsis thaliana 298-333 DRDYGILNKV FHNITDTHVA HHLFSTMPHY NAMEAT Glycine max 294-329 DRDYGILNKV FHHITDTHVA HHLFSTMPHY HAMEAT Zea mays 305-340 HHLFSTMPHY DRDYGILNRV FHNITDTHVA HAMEAT Ricinus communis3 198-224 DRDYGILNKV FHNITDTQVA HHLF TMP Brassica napus D 299-334 DRDYGILNKV FHNITDTHVA HHLFSTMPHY HAMEAT Brassica napus F 299-334 DRDYGILNKV FHNITDTHVA HHLFSTMPHY HAMEAT of pRF2- 1C plasmid TABLE 4 Alignment of Conserved Amino Acids of Fatty Acid Microsomal Desaturases Delta-12 Species Position Amino Acid Sequence Arabidopsis thaliana 165-180 IKWYGKYLNN PLGRIM Glycine max 161-176 VAWFSLYLNN PLGRAV Zea mays 172-187 PWYTPYVYNN PVGRW Ricinus communis3 65-80 IRWYSKYLNN PPGRIM Brassica napus D 165-180 IKWYGKYLNN PLGRTV Brassica napus F 165-180 IKWYGKYLNN PLGRTV of pRF2-1C plasmid TABLE 5 Alignment of Conserved Amino Acids of Plastid Desaturases v Microsomal of Fatty Acid Delta-1 5 Species Position Amino Acid Sequence Arabidopsis thaliana 3 156-177 WALFVLGHD CGHGSFSNDP KLN Brassica napus at 114-135 WALFVLGHD CGHGSFSNDP RLN Glycine max3 164-185 WALFVLGHD CGHGSFSNNS KLN Arabidopsis thaliana 94-115 WAIFVLGHD CGHGSFSDIP LLN Brassica napus 87-109 WALFVLGHD CGHGSFSNDP RLN Glycine max 93 -114 WALFVLGHD CGHGSFSDSP PLN a Plastid Sequences TABLE 6 Alignment of Conserved Amino Acids of Plaatid Desaturases v Microsomal of Fatty Acid Delta-15 Species Position Amino Acid Sequence A. thaliana3 188-216 ILVPYHGWRI SHRTHHQNHG HVENDESWH B. napus3 146-174 ILVPYHGWRI SHRTHHQNHG HVENDESWH Glycine max3 196-224 ILVPYHGWRI SHRTHHQHHG HAENDESWH A. thaliana 126-154 ILVPYHGWRI SHRTHHQNHG HVENDESWV Brassica napus 117-145 ILVPYHGWRI SHRTHHQNHG HVENDESWV Glycine max 125-153 ILVPYHGWRI SHRTHHQNHG HIEKDESWV Plastid sequences The conservation of the amino acid motifs and their relative positions indicates that the regions of the delta-12 or delta-15 fatty acid desaturase can be mutated in a species to generate a non-functional desaturase that it can be mutated in the corresponding region of other species to generate a product of the delta-12 desaturase gene or della-1 5 desaturase in that species. Mutations in any of the regions of Tables 1 through 6 are specifically included within the scope of the invention and are substantially idyllic to those mutations exemplified therein, provided that said mutation (or mutations) produces the product. of non-functional non-functional gene results, as previously described. A fragment of nucleic acid containing a mutant sequence can be generated by techniques known to those skilled in the art. Such techniques include, without limitation, site-directed mutagenesis of wild-type sequences and direct synthesis using automated DNA synthesizers. A nucleic acid fragment that contains a mutagenic sequence can also be generated by mutagenesis of plant seeds or regenerative tissues of plants with, for example, ethyl methane sulfonate, X-rays or other mutagenes. With mutagenesis, mutant plants having the desired fatty acid phenotype in the seeds are identified by known techniques, and a nucleic acid fragment containing the desired mutation is isolated from the genomic DNA or RNA of the mutant line. The site of the specific mutation is then determined, being the sequence of the coding region of the delta-12 desaturase gene or the delta-15 desaturase. Alternatively, labeled nucleic acid samples that are specific for the desired mutation events can be used to rapidly classify a mutagenized population. The described method can be applied to all the Brassica species of aceile seed, and to the ripening types in the Spring as in the Winter within each species. Physical mutagenes, including but not limited to X-rays, UV rays, and other physical disorders, which cause damage to the chromosome, and other chemical mutagenes, including but not limited to etidium bromide, nitrosoguanidine, diepoxybutane, etc. they can also be used to induce mutations. The mutagenesis tracing can also be applied to other stages of plant development, including but not limited to cell cultures, embryos, microsporos and apices of the shoots. "Stable mutations" as used in this invention are defined as M5 or more advanced lines which maintain an altered profile selected from fatty acid for a minimum of three generations, including a minimum of two generations under field conditions, and which exceed established thresholds for a minimum of two generations, as determined by the gas chromatographic analysis of a minimum of 10 randomly selected seeds grouped together. Alternatively, stability can be measured in the same way by comparing subsequent generations. In subsequent generations, esiability is defined as having similar fatty acid profiles in the seed to those of previous or subsequent generations when they are grown under substantially similar conditions. Reproduction of the mutation has traditionally produced plants that carry, in addition to the characteristic of interest, multiple, harmful characteristics, for example, a reduced vigor of the plant and reduced fertility. These characteristics may indirectly affect the fatty acid composition, producing an unstable mutation; and / or reduce production, thereby reducing the commercial utility of the invention. To eliminate the occurrence of harmful mutations and reduce the burden of mutations carried by the plant, a small dose of mutagen was used in seed treatments to create an LD30 population. This allows rapid selection of single-gene mutations for fatty acid characteristics in agronomic experiences that produce acceptable yields. The seeds of different fatty acid lines have been deposited with the American Type Culture Collection and the following access numbers are available.Access Line No. Deposit Date A129.5 4081 1 May 25 of 1990 A133.1 40812 May 25 of 1 990 M3032.1 75021 June 7 of 1991 M3062.8 75025 June 7 of 1 991 M3028.10 75026 June 7 of 1991 IMC130 75446 April 16, 1993 Q4275 97569 May 10, 1996 More than one form of endogenous delta-12 microsomal desaturas can be found in some species or plant varieties. In the amididloid, each form can be derived from one of the genomes of origin that form the species under consideration. Plants with mutations in both forms have a fatty acid profile that is different from plants with a mutation in only one form. An example of said plant is line Q508 of the Brassica napus, a double mutagenized line containing a mutant form of the desaturase della-12 (IDENTIFICATION OF SEQUENCE NO: 3) and a mulenle F form of the delta-12 desaturase (IDENTIFICATION DE SECU ENCIA NO: 7). Another example is line Q4275, which contains a mutant form D of the delta-12 desaturase (SEQUENCE IDENTIFICATION NO: 3) and a mutant form F of the delta-12 desaturase (SEQUENCE IDENTIFICATION NO: 9). See Figures 2 and 3. The preferred host organisms or recipients for the introduction of a nucleic acid fragment of the invention are the oil-producing species, such as soybeans (Glycine max), rapeseed (for example, Brassica napus) , B. rapa and B. júncea), sunflower (Helianthus annus), beaver bean (Ricinus communis), corn (Zea mays), and saffron (Carthamus tinctorius). A nucleic acid fragment of the invention may additionally comprise additional nucleic acids. For example, a nucleic acid encoding a secretory or leader amino acid sequence can be linked to a mutant fragment of desaturase nucleic acid so that the secretory or leader sequence is fused to the structure at the amino terminus of a mutant desaturase polypepide. delta-12 or delta-15. Other fragments of nucleic acid are known in the art which encode amino acid sequences useful for binding them in the structure to the desaturase mutant polypeptides described in the present document. See, for example, US Pat. No. 5,629, 193 incorporated herein by reference. A fragment of nucleic acid may also have one or more regulatory elements operably linked thereto. The present invention also comprises fragments of nucleic acid that selectively hybridize to mutant desaturase sequences. Said nucleic acid fragments generally have a length of at least 15 nucleotides. Typical hybridization comprises Southern analysis (Southern blots), a method whereby the presence of DNA sequences in a target nucleic acid mixture is identified by hybridization to a sample of a DNA fragment or labeled oligonucleotide. Southern analysis generally involves electrophoretic separation of DNA digests on agarose gels, denaturation of DNA after electrophoretic separation, and transfer of DNA to nitrocellulose, nylon, or other membranes soporles suitable for analysis. with a radiolabelled sample, biolinylated or labeled by enzyme lal and as described in Sections 9.37 to 9.52 of the study by Sambrook et al, (1989) Molecular Cloning, second edition, Cold Spring Harbor Laboratory, Plainview; NY. A nucleic acid fragment can hybridize under conditions of moderate stringency or, preferably, under conditions of high stringency to a mutant desaturase sequence. High severity conditions are used to identify nucleic acids that have a high degree of homology with the sample. High severity conditions may include the use of low ionic strength and high temperature for washing, eg, sodium citrate 0.015 M NaCl / 0.001 5 M (0.1 X SSC); 0.1% sodium lauryl sulfate (SDS) at a temperature of 50 to 65 ° C. Alternately, a denaturing agent such as formamide can be employed during hybridization, for example, 50% formamide with 0.1% bovine serum albumin / 0.1% Ficoll / 0.1% polyvinylpyrrolidone / 50 mM sodium phosphate buffer at a pH of 6.5 with 750 mM NaCl, 75 mM sodium citrate at a temperature of 42 ° C. Another example is the use of 50% formamide, 5 x SSC (0.75 M NaCl, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1% sodium pyrophosphate, 5 x Denhardt's solution, DNA of sonicated salmon sperm (50 μg / ml), 0.1% of SDS, and 10% of dextran sulfate at a temperature of 42 ° C, with washes at temperatures of 42 ° C in 0.2 x SSC and 0.1% SDS. The conditions of moderate severity refer to hybridization conditions used to identify nucleic acids that have a lower degree of identity with the samples of the nucleic acids identified under conditions of high severity. Conditions of moderate severity may include the use of higher ionic strength and / or lower temperatures for washing the hybridization membrane, compared to the ionic strength and temperatures used for high stringency hybridization. For example, a wash solution comprising 0.060 M NaCl / 0.0060 M sodium citrate (4X SSC) and 0.1% sodium lauryl sulfate (SDS) at a temperature of 50 ° C can be used. With a final wash in 1 X SSC at a temperature of 65 ° C. Alternatively, a hybridization wash in 1 X SSC can be used at a temperature of 37 ° C. Hybridization can also be done through Norie's analysis (Northern blots), a method used to identify ARNS that hybridize a known sample such as an oligonucleotide, DNA fragment, cDNA or fragments thereof, or an RNA fragment. The sample is labeled with a radioisotope such as 32P, by biotinylation or as an enzyme. The RNA to be analyzed can normally be separated by electrophoresis in an agarose or polyacrylamide gel, transferred to microcellulose, nylon or another suitable membrane and hybridized with the sample, using standard techniques well known in the art, such as which are described in sections 7.39 to 7.52 of Sanbruc on, above. A polypeptide of the invention comprises an isolated polypeptide having a mutant amino acid sequence, as well as derivatives and analogs thereof. See for example, the amino acid mutant sequences of Figure 3. The term "isolated" means a polypeptide that is expressed and produced in an environment different from the environment in which the polypeptide is naturally expressed or produced. For example, a plant polypeptide is isolated when expressed and produced in bacteria or fungi. A polypeptide of the invention also comprises variants of the mutant desaturase polypeptides described in this invention, as explained above.

In one embodiment of the claimed invention a plant contains both a delta-12 desaturase mutation and a delta-1 5 desaturase mutation. Said plants can have a fatty acid composition comprising very high levels of oleic acid and very low levels of alpha linolenic acid. Mutations in delta-12 desaturase and della-15 deaceurase can be combined in a plant by making a genetic crossing in simple mutant lines between della-12 desaturase and delta-15 desaturase. A plant that has a mutation in the delta-12 fatty acid desaturase is crossed or paired with a second plant that has a mutation in the delta-1 5 fatty acid desaturase. The seeds produced from the cross are planted and the resulting plants are self-pollinated in order to obtain seeds of descent. These seeds of the descendants are then selected in order to identify those seeds that carry both mutant genes. Alternatively, a line having a mutation, either delta-12 desaturase or delta-1 5 desaturase may be subjected to mulagenesis to generate a plant or line of plants that has mutations in both delta-12 and delta-12 the delía-15 desaturase. For example, the IMC 129 line has a mulation in the coding region (GLU106 to Lis106) of the D form of the microsomal structural gene of the delta-12 desaturase. The cells (e.g., seeds) of this line can be mutagenized to induce a mutation in a delta-15 desaturase gene resulting in a plant or line of plants carrying a mutation in a delta-fatty acid desaturase gene. 12 and a mutation in a delia-15 fatty acid desaturase gene. The offspring include the descendants of a particular plant or plant line, for example, the seeds developed in a present plant are the descendants. The offspring of a présenle plant includes seeds formed in F? , F2, F3, and the plants of the subsequent generation, or seeds formed in BCi, BC2, BC3 and plants of the subsequent generation. The plants according to the invention preferably contain an altered fatty acid composition. For example, the oil obtained from the seeds of said plants may have from about 69% to about 90% oleic acid based on the total fatty acid composition of the seed. Said aceyl preferably has from about 74% to about 90% oleic acid, more preferably from about 80% to about 90% oleic acid. In some embodiments, the oil obtained from the seeds produced by plants of the invention may be from about 2.0% to about 5.0% saturated fatty acids, based on the total fatty acid composition of the seeds. In some embodiments, the oil obtained from the seeds of the invention may have from about 1.0% to about 14.0% linoleic acid, or from about 0.5% or up to about 10.0% a-linolenic acid. The composition of the oil is usually analyzed by grinding and extracting the fatty acids from the samples of several seeds (for example, 10 seeds). The triglycerides of the fatty acid in the seed are hydrolyzed and converted into methyl esters of fatty acid. Those seeds that have an altered fatty acid composition can be identified by techniques known to those skilled in the art, for example, gas-liquid chromatography (GLC) analysis of a sample of several seeds, a single seed, or a only half seed The analysis of the half seed is well known in the art to be useful because the viability of the embryo is maintained and thus, those seeds having a desired fatty acid profile can be planted to form the next generation. However, it is also known that the analysis of half seed is an inaccurate representation of the genotype of the seed that is being analyzed. The analysis of several seeds generally produces a more accurate representation of the fatty acid profile of a given genotype. The analysis of half seed of a population of seeds is, however, a reliable indicator of the probability to obtain a desired profile of fatty acid. The fatty acid composition can also be determined in larger samples, for example, the oil obtained by pilolo plants, or the commercial scale, bleached, and deodorized refining of the endogenous oil in the seeds. The nucleic acid fragments of the invention can be used as markers in the genetic tracing of plants, and plant breeding programs. Such markers may include a restriction length polymorphism of the fragment (RFLP), a random polymorphism detection of amplification (RAPD), a polymerase chain reaction (PCR) or self-sustained sequence reproduction markers (3SR), for example.

The culíivo techniques aided by markers can be used to identify and follow a desired composition of fatty acid during the culture process. The culture techniques aided by the markers can be used in addition to, or as alternatives to, other IDENTIFICATION classification techniques. An example of marker assisted culture is the use of PCR primers that specifically amplify a sequence containing a desired mutation in delta-12 desaturase or delta-1 desalurase 5. The methods according to the invention are useful because the resulting plants and the plant lines have desirable fatty acid compositions in the seeds, as well as superior agronomic properties compared to known lines which have altered fatty acid compositions in the seeds. The superior agronomic characteristics include, for example, an increased percentage of seed germination, an increased seedling vigor, an increased resistance to fungal diseases of the seedling (amorliguing, root puírefaction and the like), increased production and improved stability. In another aspect, Brassica plants are characterized which produce seeds having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% based on the total composition of fatty acid. As used in this invention, "long chains" refers to carbon chains of 16 and greater, for example chains of 16 to 24 carbons. The content of long chain monounsaturated fatty acid is distributed mainly between oleic acid, eicosenoic acid and erucic acid. The heterogeneous nature of the long chain monounsaturated fatty acids in the triacylglycerols of the seed oil confers desirable oil properties. The levels of total saturated fatty acids and / or total polyunsaturated fatty acids can be decreased in order to increase the long chain monounsaturated content, for example, oleic acid, eicosenoic acid, erucic acid and nervonic acid. The high oleic acid lines described here can be crossed with high erucic acid lines to produce Brassica flavors that have a long chain monounsaturated fatty acid condense within their seeds. Suitable high oleic acid lines are described, as for example, in Example 5 and Table 17, and have an oleic acid content of about 82% up to about 85%, based on the total fatty acid composition. Suitable elevated erucic acid lines have an erucic acid content of about 45% or higher, based on the total fatty acid composition. The Brassica plant HEC01 line is an elevated erucic acid line that is particularly useful and sold under the Hero trademark. Other varieties of high erucic acid are well known, such as the varieties designated Venus, Mercury, Neptune, S89-3673, Dwarf Essex, Reston, Bridger or R-500. P. B.E. McVeíty et al, in Can. J. Plant Sci., 76 (2): pages 341-342 (1996); the study by R. Scaríh el al, in Can J. Plant Sci., 75 (1): pages 205-206 (1,995); and the study of P. B. E. Me Vetty el al, in Can. J. Plañí Sci., 76 (2): pages 343-344 (1996). The seeds of the invention may have content of oleic acid and eicosenoic acid of at least about 37% and 14% respectively, based on the total fatty acid composition. The total content of saturated fatty acid is less than approximately 7%. As used in this description, "tofal content of safed fatty acid" refers to the myristate (14: 0), palmitate (1 6: 0), stearate (18: 0), arachididate (20: 0), behenate (22: 0) and lignocerate (24: 0). The total polyunsaturated content is less than about 1 1% based on the total fatty acid composition. As used in this invention"Total polyunsaturated fatty acid connate" refers to the sum of linoleic (18: 2), a-linolenic (18: 3) and eicosadienoic (20: 2) fatty acids as a percentage of the total fatty acid content. In some embodiments, the long chain monounsaturated content in the seeds is from about 85% to 90%. The content of oleic acid within these seeds is about 42% or greater, and preferably about 47% >; to approximately 56%. The content of erucic acid and eicosenoic acid are from about 17% to about 31% > and from about 15% to about 21% > , respectively. In some embodiments, the long chain monounsaturated content in the seeds is from about 82% to about 90%. The content of oleic acid in these seeds is approximately 25% up to 33%. The conlicides of erucic acid and eicosenoic acid are from about 44% to about 50% and from about 10% to about 13%, respectively. Seed oils having a monounsaturated long-chain confenide of at least about 82% and an erucic acid content of at least about 15%, based on the total fatty acid composition, were also characterized. These oils can be extracted, for example, from a single line of Brassica seeds having a suitable fatty acid composition as described in this invention. The content of oleic acid and eicosenoic acid of these oils is at least about 37% and 14%, respectively, based on the total fatty acid composition. The total saturated and polyunsaturated content of these oils is less than about 7% and 11%, respectively. Preferably, the polyunsaturated content is less than about 9%. In some embodiments, the oils have a monounsaturated content of about 85% to about 90%. The oleic acid content of these oils is at least about 42% and more preferably about 47% > to approximately 56%. Aceifes having an erucic acid content of about 17% to about 31% and an eicosoic acid content of about 15% to about 21% > . In some embodiments, the long chain monounsaturated content in said oils is about 82% up to about 90%, comprising an oleic acid content of about 25% to about 33%, an erucic acid content of about 44% to about 50% > and an eicosenoic acid content of about 10% up to about 13%. The sum of the contents of nervonic acid, erucic acid and eicosenoic acid in said oils may be from about 50% to about 66%. Alternatively, it is contemplated that the oils of the invention may be obtained by mixing rape seed oil with a high content of erucic acid (H EAR) and an oil having at least about 87% oleic acid, preferably from about 90% to about 95% oleic acid, based on the total fatty acid composition. The HEAR oil has an erucic acid content of approximately 49% and an oleic acid content of approximately 16%. Oils having a long chain monounsaturated content of at least about 82%, unexpectedly have low temperature properties which are desirable for industrial applications such as lubrication. The basis of these properties is not known, but it is possible that the lengths of the heterogeneous chains of the triacylglycerols in the oils of the invention orderly prevent packaging as final methyl groups that have a discordance in the molecular volume, reducing Van interactions. der Waals. The double bond in each fatty acid moiety is present at different carbon positions along the acyl chain, which can hinder packaging and also reduce electronic p-p interactions between adjacent fatty acid chains. The elevated monounsaturated conure is considered to provide improved oxidative stability together with high flow characteristics. Low levels of polyunsaturated oils in the invention also promote high oxidative stability, because the oxidation rates of linoleic acid and linolenic acid at temperatures of 20 ° C are 12 to 20 times and 25 times, respectively, higher than the oxidation rate of oleic acid. Oxidative stability can be measured with an Oxidative Stability Index Instrument instrument, Omnion, Inc., Rockland, MA, according to the AOCS Official Method Cd 12b-92 (revised in 1993). The method is an automatic replacement of the procedure of the active oxygen method (AOM) Official Method AOCS Cd 12-57. The oxidative stability of oils containing long chain monounsaturated content of at least about 82% >; it is about 40 AOM hours up to about 100 AOM hours in the absence of added antioxidants. In comparison, medium-oleic cañola oil (approximately 76% oleic acid) and high erucic acid rapeseed oil have oxidative stabilities of approximately 38 and 16 hours AOM, respectively, in the absence of added antioxidants. The oils of the invention have desirable functional properties, for example low-temperature compression and a high viscosity index, together with high oxidative stability. The presence of higher molecular weight fatty acids increases the molecular weight of the triacylglycerols, producing the oil with a higher flash point, and a higher ignition point. The increased molecular weight also improves the viscosity index of the oils. The viscosity index is an arbitrary number that indicates the change in viscosity with the temperature of a lubricant. The viscosity index of Dean and Davis can be calculated from the observed viscosities of a lubricant at a lemperaiura of 40 ° C and 100 ° C and can produce values in a range from 0 to values greater than 200. A higher value of the index of indica viscosity that the viscosity of the oil changes less with a change in temperature. In other words, at a higher viscosity index, the difference in viscosity between high and low temperatures is lower. An oil of the invention can be formulated for industrial applications such as engine lubricants or hydraulic fluids by the addition of one or more additives to an oil having a long chain monounsaturated fatty acid content of at least about 82%. and an erucic acid conifer of at least about 15%, based on the total fatty acid composition. For example, a liquid can be made for the transmission of diesel engines that includes antioxidants, anti-foam additives, anti-wear additives, corrosion inhibitors, dispersants, acid detergents and neutralizers or combinations thereof. The hydraulic oil compositions may include antioxidants, anti-rust additives, anti-wear additives, pour point depressants, viscosity index improvers and anti-foam additives or combinations thereof. The specific formulations will vary depending on the final use of the oil; The adaptability of a formulation for a final use can be evaluated using standard techniques. Typical antioxidants include zinc dithiophosphates, methyl dithiocarbamates, hindered phenols, phenol sulphides, phenol metal sulfides, metal salicylates, aromatic amines, phospho-sulfurized fats and olefins, sulfurized olefins, sulfurized fats, and fatty derivatives, sulfurized paraffins. , sulfurized carboxylic acids, disalieilal-1,2, -propane diamine, 2,4-bis (alkyldiio-1, 3,4-thiadiazoles) and dilauryl selenide. Antioxidants are generally present in amounts of about 0.01% to about 5% based on the weight of the composition. In particular, it is added to an oil of the invention of about 0.01% or up to about 1.0% of antioxidant. See U.S. Patent No. 5,451, 334 for additional antioxidants. Rust inhibitors protect the surfaces against rust and include, for example, organic acids of the alkylsuccinic type and derivatives thereof, alkylthioacetic acids and derivatives thereof, organic amines, organic phosphates, polyhydric alcohols and sodium and calcium sulfonates. The anti-wear additives are absorbed in the metal and produce a film that reduces the metal to metal content. In general, anti-degassing additives include zinc dialkyldithiophosphate, tricresyl phosphate, didodecyl phosphite, sulfurized sperm oil, sulfurized terpenes, and zinc dialkydithiocarbamate, and are used in amounts of approximately 0.05% >; up to about 4.5%. Corrosion inhibitors include dithiophosphates and in particular zinc dithiophosphates, metal sulfonates, metal phenate sulphides, fatty acids, acid phosphate esters, and alkylsuccinic acids. The pour point depressors allow the flow of oil composition below the pour point of an unmodified lubricant. Common spill depressants include polymethacrylates, alkylated wax naphthalene polymers, alkylated wax phenol polymers, and chlorinated polymers and are generally present in amounts of about 1% or less. See, for example, US Patents Nos. 5,451, 334 and 5,41 3,725. The viscosity index can be increased by adding polybutybutylenes, polymethacrylates, polyacrylates, ethylene propylene copolymers, styrene-isoprene copolymers, styrene-butadiene copolymers and styrene-maleic esters. The anti-foam additives reduce or prevent the formation of a stable foam on the surface and are generally present in amounts of about 0.00003% to about 0.05%. Polymethylsiloxanes, polymethacrylates, alkyl alkylene dithiophosphate salts, amyl acrylate telomers and poly (2-ethylexylacrylate-co-ethyl acrylate) are non-limiting examples of the anti-foam additives. Detergents and dispersants are polar materials that provide a cleaning function. Detergents include metal sulfonates, metal salicylates and metal thiophosphonates. Dispersants include polyamines succinimides, polyamines hydroxybenzyl, succinimides polyamine, polyhydroxy succinic esters and polyamine amide imidazolines. Although the present invention is suscepible of various modifications and shapes, all specific embodiments thereof are described in the general methods and examples presented below. The present invention can be applied to all Brassica species including β. Rapa, B.juncea, and B. Hirta, to produce substantially similar results. However, it should be understood that these examples are not intended to limit the invention to the paricular forms described herein, but on the contrary, the present invention covers all modifications, equivalents and alternatives that are within the scope thereof. This includes the use of somaclonal variations; physical and chemical mutagenesis of parts of plants; cultures of anther, microsporum and ovaries followed by the duplication of chromosomes; self-pollination and cross-pollination to transmit the characteristics of the fatty acid, alone or in combination with other characteristics, to develop new Brassica lines.

EXAMPLE 1 Muíaaénesis The seeds of Weslar, a spring variety of the Canadian cañola (Brassica napus), are somelidas to the chemical mutagenesis. The Westar is a Canadian spring registered variety of cañola quality. The fatty acid composition of Westar cultivated in the field, 3.9% Ci ß: o. 1 -9% C? ß: o. 67.5% C? 8: 1, 17.6% C18: 2, 7.4% C18: 3, < 2% C20: 1 + C22: 1, has remained stable under commercial production, as a deviation from < ± 10% since 1982. Before mulagénesis, 30,000 seeds of B. Napus cv. Westar seeds were previously impregnated in batches of 300 seeds for two hours on a wet filter paper to soften the seed shell. The previously impregnated seeds were placed in 80 mM ethylmeanosulfonate (EMS) for four hours. After mutagenesis, the seeds were rinsed three times in distilled water. The seeds were planted in trays of 48 containers containing Pro-Mix. 68%) of the mulagenized seeds germinated. The plants were maintained at an emperature of 25 ° C / 15 ° C, during conditions of 14/10 hours / day / night in the greenhouse. Upon flowering, each of the plants was self-pollinated individually. The M2 seed of the individual plants were cataloged and stored individually, lines of approximately 15,000 M2 were planted in a summer infirmary in Carman, Manitoba. The seeds of each of the self-pollinated plants were planted in rows of 3 meters with a space between the rows of 6 inches, the Westar was planted as the variety for revision. The selected lines in the field were self-polishing, covering with bags the main cluster of each plant. At ripening, the self-pollinated plants were harvested individually and the seeds were cataloged and stored to ensure that the origin of the seeds was known. The M3 self-pollination seeds and Wesfar controls were analyzed in groups of 10 seeds to determine the fatty acid composition by means of gas chromatography. Statistical thresholds for each fatty acid component were established using a Z-distribution with a severity level of one in 10,000. The average values of the standard deviation and average were determined from the control population of Westar not mutagenized in the field. The upper and lower statistical thresholds for each fatty acid were determined from the average value of the population ± the standard deviation, multiplied by the Z-distribution. Based on a population size of 10,000, the confidence interval is 99.99%.

The selected M3 seeds were planted in the greenhouse together with the Westar controls. The seeds were sown in 9.16 centimeter containers that contain the Pro-Míx and the plants were maintained at a temperature of 25 ° C / 1 5 ° C, in a day / night cycle of 14/1 0 hours in the greenhouse. As they bloom, the final clusters were self-pollinated inside bags. Upon maturation, the self-pollinated M4 seeds of each plant were harvested individually, labeled and stored to ensure that the origin of the seed was known. The M4 seed was analyzed in samples of groups of 10 seeds. The statistical thresholds for each fatty acid component were established from 259 samples of conírol using a Z-distribution of one in 800. The selected M lines were planned in a field trial in Carman, Maniloba, in rows of 3 meters and with a separation of 6 inches. Ten M plants in each row were covered with bags for self-pollination. Upon maturation, the self-pollinated plants were harvested individually and the open pollen plants of the row were harvested by bulk volume. The M5 seed of the single-plant selections were analyzed in samples of groups of 10 seeds, and the harvest of the row in bulk in samples of groups of 50 seeds. The selected M5 lines were planted in the greenhouse along with the Weslar controls. The seeds were cultivated as described above. When blooming, the terminal cluster was self-pollinated in bags. Upon maturation, the self-pollinated M6 seeds of each plant were harvested individually and analyzed in samples of groups of 10 seeds to determine the fatty acid composition.

The selected M6 lines were introduced in field trials in East Idaho. The four locations of the trials were selected for the wide variability in culture conditions. Locations included Burley, Tetonia, Lamont and Shelley (Table 7). The lines were planned in rows of 3 meters with a separation of 8 inches, each row was reproduced four times. The plantation design was determined using a complete block design in a random manner. The commercial cultivation Westar was used as a revision crop. At maturity, the lines were collected to determine production. The production of the introduction in the trial was determined by taking a statistical average of the four reproductions. The least significant difference test was used to classify the entries in the completely randomized block design.

TABLE 7 Test Locations for Selected Fatty Acid Mutants. LOCATION SITE FEATURES BURLEY Irrigated. Long season, high temperaluras duraníe el Blossoming TETONIA Dry land, short exposure, cold temperatures LAMONT Dry land, short season, cold temperatures SHELLEY Irrigated, station measured high temperatures during flowering In order to deveer the fatty acid profile of these iníroducciones, the plants of each of the lines were covered with bags for self-pollination. The M7 seed of single plants was analyzed to determine the fatty acids in samples of groups of 10 seeds. To determine the genetic relationships, crosses of selected fatty acid mutants were made. The flowers of M6 or mutations of later generations were used in the crossing. The seed F ^ was harvested and its fatty acid composition analyzed to determine the mode of action of the gene. The offspring of F- [was planted in the greenhouse. The resulting plants were self-pollinated, the F2 seed was harvested and their fatty acid composition was analyzed for allelism studies. The seed F2 and the seed of the line of origin were planted in the greenhouse, the individual plants were auoproced. The F3 seed of the individual seeds was tested to determine the fatty acid composition using samples of groups of 10 seeds as described above. In the analysis of some genetic relationships, diaploid populations were made from microspores of Fi hybrids. The self-pollinated seeds of diaploid plants were analyzed for their fatty acid content using the methods described above. For the chemical analysis, samples of groups of 10 seeds were sown by hand with a glass rod in a 15 ml polypropylene tube. and extracted in 1.2 ml. of KOH 0.25 N in 1: 1 ether / methanol. The samples were stirred to one side for 30 seconds and heated for 60 seconds in a water bath at 60 ° C. 4 ml was added. of saturated NaCl and 2.4 ml. of so-octane and the mixture was agitated again to one side only. After phase separation, 600 μL of the upper organic phase were packed in individual vials and stored under nitrogen at a temperature of -5 ° C. One μL samples were injected into a Supelco SP-2330 fused silica capillary column (0.25 mm ID, 30 M length, 0.20 μm df). Gas chromatography was adjusted to 1 80 ° C for 5.5 minutes and then programmed for an increase of 2 ° C per minute to 212 ° C and was held at this temperature for 1.5 minutes. The total time of the process was 23 minutes. Chromatography settings were: column head pressure-15 psi, column flow (He) - 0.7 mL / min, column and auxiliary flow - 33 mL / min, hydrogen flow - 33 mL / min , air flow - 400 mL / min, injector reading -250 ° C deteclor temperature - 300 ° C, split vent - 1/15. Table 8 describes the upper and lower statistical upper levels for each fatty acid of interest.

TABLE 8 Statistical Thresholds for Specific Fatty Acids Derived from Westar Control Plantations Percentage of Fatty Acids Genotype '16: 0'18: 0 C18; C18: C18: 3 Sats * Generation M3 (Rejection index 1 In 10,000) Inferior 3.3 1.4 - 13.2 5.3 6.0 Superior 4.3 2.5 71.0 21.6 9.9 8.3 Generation M4 (Rejection index 1 In 800) Inferior 3.6 0.8 - 12.2 3.2 5.3 Superior 6.3 3.1 76.0 32.4 9.9 11.2 Generation M5 (Rejection rate 1 In 755) Inferior 2.7 0.9 - 9.6 2.6 4.5 Superior 5.7 21 803 26.7 9.6 10.0 EXAMPLE 2 Lines of Cañola of High Oleic Acid. In the studies of Example 1, in the M3 generation, 31 lines exceeded the upper statistical threshold for oleic acid (= 71.0%). Line W7608.3 had 71.2% oleic acid. In Generation M, their self-pollinated offspring (W7608.3.5, designated A129.5) conjoined exceeding the upper statistical threshold for C? 8:? with 78.8% oleic acid. The M5 seed of five self-pollinated plants of line A129.5 (ATCC 4081 1) averaged 75.0% oleic acid. A single-plant selection, A129.5.3 had 75.6% oleic acid. The fatty acid composition of this high mutanle in oleic acid, which was stable, both under field conditions and in the greenhouse for an M7 generation, is summarized in Table 9. This line also remained stable in its mutant composition. fatty acid for the M7 generation in the field trials in multiple locations. In all locations, self-pollinated plants (A129) averaged 78.3% oleic acid. The fatty acid composition of A129 for each Idaho test location is summarized in Table 1 0. In replicated field trials of multiple locations, A129 was not significantly different in Westar production from home culture. Cañola oil from A129, after commercial processing, was found to have superior oxidative stability compared to Westar when measured by the Accelerated Oxygen Method (AOM), American Oil Chemists' Society Official Method Cd 12-57 for stability of fats; Active oxygen method (revised in 1989). The AOM of the Westar was 1 8 hours AOM and for the A129 it was 30 AOM hours. TABLE 9 Fatty Acid Composition of Oleic Acid Canopy Line The Evade or Production of Seed Mutagenesis.

Percentage of Fatty Acids Genotype Cl6: 0 C? S: 0 Cl8: 1 C-I8: 2 Ci8: 3 Sats Westar 3.9 1.9 67.5 17.6 7.4 7.0 W7608.3 3.9 2.4 71.2 12.7 6.1 7.6 (M3) W7608.3.5 3.9 2.0 78.8 7.7 3.9 7.3 (M4) A129.5.3 3.8 2.3 75.6 9.5 4.9 7.6 (5) Saturated Sats = Total saturated content.

TABLE 10 Fatty Acid Composition of an Elevated Oleic Mutant Line at Different Field Locations in Idaho.

Percentage of Fatty Acids Location ^ -iß? < - 8: 0 u18: 1 C18: 2 '-'Iß ^ Sats Burley 3.3 2.1 77.5 8.1 6.0 6.5 Tetonia 3.5 3.4 77.8 6.5 4.7 8.5 Lamont 3.4 1.9 77.8 7.4 6.5 6.3 Shelley 3.3 2.6 80.0 5.7 4.5 7.7 Sats = Total saturated content. The genetic relationship of the A129 mutation of high oleic acid with oleic desaturases was demonstrated in the crossings made to commercial cane crops, and a low mutation of linolenic acid. The A129 was crossed with the global commercial crop (C16: o - 4.5%, C 8: or 1.5%, C18: --62.9%, C18: 2 - 20.0%, C18: 3 - 7.3%). The fatty acid composition of about 200 individual F2 was analyzed. The results are summarized in Table 1 1. The 1: 2: 1 ratio of segregation adjustment suggests a single dominant gene conflated in the inheritance of the elevated oleic acid phenotype. TABLE 1 1 Genetic Studies of A129 X Global Frequency C18: 1 Genotype (%) of Content Observed Expected od-od- 77.3 43 47 od-od + 71.7 106 94 od + od + 66.1 49 47 A cross was made between A129 and IMC 01, a variety low in linolenic acid (C16: 0 - 4.1%, C? 8: 0 - 1 .9%, C18: 1 - 66.4%, C18: 2 - 18.1%, C18: 3 -5.7%), to determine the inheritance of the oleic acid desaturase and the linoleic acid desaturase. In the F ^ tanio hybrids, the desalurase genes of oleic acid and linoleic acid approximated the mean values of origin that indicate a co-dominant action of the gene. The fatty acid analysis of the F2 individuals confirmed a 1: 2: 1: 2: 4: 2: 1: 2: 1 segregation of two independent co-dominant genes (Table 12). A crossing line of A1 29 and IMC01 was selected and designated as I MC130 (ATCC deposit No. 75446), as described in the Application of North American Patent No. 08/425, 108, incorporated herein by reference.

TABLE 12 Genetic Studies of A129 X IMC 01 Frequency Frequency Genotype Proportion Observed Expected od-od-ld-ld- 1 11 12 od-od-ld-ld + 2 30 24 od-od-ld + ld + 1 10 12 od-od + Id-ld- 2 25 24 od-od + ld-ld + 4 54 47 od-od + ld + ld + 2 18 24 od + od + ld-ld- 1 7 12 od + od + ld-ld + 2 25 24 od + od + ld + ld + 1 8 12 It was also produced by the method described, an additional line of high oleic acid, designated A128.3,. An analysis of a group of 50 seeds of this line showed the following fatty acid composition: C16: 0-3.5%, C18: 0-1.8%, C18; 1 - 77.3%, C18: 2 - 9.0%, C18: 3 - 5.6%, saturated FDA - 5.3%, total saturated - 6.4%. This line also kept its mutant fatty acid composition stable for the generation of M7. The trials in the field of duplicate multiple locations, A128.3 were not significantly different in their production from the Westar culture of origin. The A129 was crossed with the A128.3 for the alelism studies. The fatty acid composition of the F2 seed showed that two lines were allelic. The mutation events in A129 and A128.3, although of different origin, were in the same gene. An additional line of high oleic acid, designated M3028.-10 (ATCC75026), was also produced by the method described in Example 1. Analysis of a group of 1 0 seeds of this line showed the following fatty acid composition: C? 6: o - 3.5%, C18: 0 - 1 .8%, C18: 1 - 77.3%, C18: 2 - 9.0 %), C18: 3 - 5.6%, saturated FDA - 5.3%, Total Saturated - 6.4%. In a field trial replicated in a single location the M3028.10 was not significantly different from the Westar culture of origin in the field. EXAMPLE 3 Low Linoleic Acid Cañola In the studies of Example 1, in generation M3 80 lines exceeded the lower statistical threshold for linoleic acid (< 13.2%). Line W12638.8 had 9.4% linoleic acid. In generations M4 and M5 the self-pollinated descendants [W12638.8, designated since then as A133.1 (ATCC 40812)] continued to exceed the statistical threshold for C18: 2 with linoleic acid levels of 10.2% and 8.4% respeclivamenle. The fatty acid composition of this low-linoleic acid mutant, which was stable for M7 generation under both field and greenhouse conditions, is summarized in Table 13. In replicated field trials in multiple locations, the A133 it was not significantly different in its production from the Westar of the crop of origin. An additional line of low linoleic acid designated M3062.8 (ATCC 75025) was also produced by the described method. An analysis of a group of 1 0 seeds of this line showed the following fatty acid composition: C? 6: or -3.8%, C18: 0-2.3%, C1 B :? - 77.1%, C18: 2 - 8.9%, C18: 3 - 4.3%, FDA certified - 6.1%). This line also remained stable in its mulenic fatty acid composition in the field and the greenhouse. TABLE 13 Acid Composition Grase > from a Line of Cañóla de Acido Linole ico Baj < o Produced by M utaq enesis of the Sem l ia.

Percentage of Fatty Acids Genotype Ci6: 0 Clß; 0 C18: 1 Cl8: 2 C? ß: 3 Uats " Westar 3.9 1.9 67.5 17.6 7.4 7.0 W12638.8 3.9 2.3 75.0 9.4 6.1 7.5 (M3) W12638.8.1 4.1 1.7 74.6 10.2 5.9 7.1 (M4) A133.1.8 3.8 2.0 77.7 8.4 5.0 7.0 (M6) aThe letters and numbers up to the second decimal point indicate the plant line. The number after the second decimal point indicates an individual plant. bSatured = Total Saturated Content.

EXAMPLE 4 Cañola with Linolenic Acid and Low Linoleic Acid In the studies of Example 1, the M3 generation? 57 lines exceeded the lower statistical threshold of linolenic acid (< 5.3%). Line W14749.8 had 5.3% linolenic acid and 15.0% linoleic acid. In generations M and M5, their self-pollinated descendants [W14749.8, since then designated M3032 (ATCC 75021)] continued to exceed the statistical threshold for C18: 3 with linolenic acid levels of 2.7% and 2.3%, respectively, for a low sum of linolenic and linoleic acids with tools of 1 1 .8% and 12.5% respectively. The fatty acid composition of this low linolenic acid, plus the mutant of linoleic acid, which was stable for the M5 generation, both in field conditions and in the greenhouse, are summarized in Table 14. In the repeated field test from a single location the M3032 was not significantly different in the production of the original crop (Westar). TABLE 14 Fatty Acid Composition of a Line of Lower Linolenic Acid Cañola Produced by Mutagenesis of the Seed - Percentage of Fatty Acids • Percentage of Fatty Acids Genotype l6: 0 Cid.o Cl8.1 C18.2 18.3 Sats Westar 3.9 1.9 67.5 17.6 7.4 7.0 W14749.8 4.0 2.5 69.4 15.0 5.3 6.5 (M3) M3032.8 3.9 2.4 77.9 9.1 2.7 6.4 (M4) M3032.1 3.5 2.8 80.0 10.2 2.3 6.5 (M5) Saturated = Total Saturated Content.

EXAMPLE 5 Lines of Cañóla Q508 v Q4275 The seeds of line B. napus IMC-129 were mutagenized with methyl N-nitrosoguanidine (MNNG). Imaging with MNNG consisted of three parts: pre-rinsing, mutagen application and washing. A phosphate regulator Sorenson 's A 0.05M was used to maintain the previous rinse, and the pH of the treatment of the mutagen in 6.1. Two hundred seeds were treated at the same time on filter paper (Whafman # 3M) in a petri dish (100mm x 15mm). The seeds were previously soaked in 15% of Sorenson's buffer 0.05M, pH 6.1, under continuous stirring for two hours. At the end of the previous soaking period, the regulator was removed from the plate. A concentration of 1 μm of MNNG was prepared in Sorenson's buffer 0.05M pH 6.1 before use. 15 ml of 10M MNNG was added to the seeds in each dish. The seeds were incubated at a temperature of 22 ° C ± 3 ° C in the dark under constant agitation for four (4) hours. At the end of the incubation period, the mutagen solution was removed. The seeds were washed with three changes of distilled water in 1 0 minute intervals. The wash was for 15 minutes. This treatment regimen produced a LD60 population. The treated seeds were planted in standard greenhouse cultivation soil and placed inside an environmentally protected greenhouse. The plants were grown under a 16-hour light regime. Upon flowering, the bunches were covered with bags to produce self-pollinated seeds. In maturation, the M2 seed was cultivated. Each M2 line was given an identification number. The population of seeds prepared with MNNG was designated as the Q series. The harvested M2 seeds were planted in the greenhouse. Growth conditions were maintained as described previously. At flowering, the clusters were covered with bags for the auopropolination. At maturation, the M3 seed was harvested and its fatty acid composition was analyzed. For each M3 seed line, approximately 10 to 15 seeds of the group were analyzed as described in Example 1.

The M3 lines of low linoleic acid and high oleic acid were selected from the M3 population using a cut of > 82% oleic acid and < 5.0% linoleic acid. Of the first 1,600 M3 lines selected for their fatty acid composition, Q508 was identified. The generation of M3 Q508 was advanced to the M4 generation in the greenhouse. Table 5 illustrates the fatty acid composition of Q508 and I MC 129. Self-pollinated M4 seeds maintained the low linoleic acid phenotype, and high oleic acid (Table 16). TABLE 15. Composition of Fatty Acid of A129 and M3 Mutant Q508 of High Oleic Acid. Line # 16: 0 18: 0 18: 1 18: 2 18: 3 A129 * 4.0 2.4 77.7 7.8 4.2 Q508 3.9 2.1 84.9 2.4 2.9 * The fatty acid composition of A129 is the average of 50 self-pollinated plants grown with a population M3.

Generation M of Q508 plants had poor agronomic qualities in the field compared to Weslar. The typical plants had a slow growth in relation to the Westar, they lacked the early vegetative vigor, they had corian stature and they were to be chlorííicas and they had corolla pods. The production of the Q508 was very slow compared to the Westar. The M4 generation of the greenhouse plants Q508 tended to have reduced vigor compared to the Westar. However, the Q508 produced in the greenhouse were larger than those produced in the Q508 field.

TABLE 16. Fatty Acid Composition of Seed Oil Q508. IMC 129 v Westar Cultivated in the Greenhouse. aPrornedio of 50 self-pollinated plants bData of Example 1 cAverage of 50 self-pollinated plants Also identified were nine different lines of M4 low in linoleic acid and high in oleic acid: Q3603, Q3733, Q4249, Q6284, Q6601, Q6761, Q7415, Q4275, and Q6676. Some of these lines had good agronomic characteristics and an elevated level of oleic acid in the seeds of approximately 80% to approximately 84%. The Q4275 was cultivated in the Cyclone variety. After the self-pollination for seven generations, the mature seed of 93GS34-179 was harvested, a line of the offspring of the Q4275xCyclone cross. Referring to Table 17, the fatty acid composition of the seed set sample shows that 93GS34 retained the fatty acid composition of seed Q4275. 93GS34-179 also maintained agronomically desirable characteristics. After more than seven generations of self-pollination of the Q4275, the plants of Q4275, I MC 129 and 93GS34 were harvested in the field during the summer season. The selections were tested on 4 repeated graphs (5 feet X 20 feet) in a randomized block design. The plants were polished in the open. And the aufopolinated seeds were produced. Each of the lines was harvested at maturity, and a bulk sample of the harvested seed from each of the lines was analyzed to determine the fatty acid composition as previously described. The fatty acid compositions of the selected lines are illustrated in Table 17.

TABLE 17 Fatty Acid Composition of IMC 129 Seeds. Q4275 and 93GS34 Harvested in the Field.

The results shown in Table 17 show that Q4275 maintained the high oleic acid phenotype - low linoleic acid under field conditions. The agronomic characteristics of the Q4275 pineapples were superior to those of the Q508. Generation M of the plans Q508 was crossed for a dihaploid selection of Westar, serving the Westar as the seed of female origin. The resulting F1 seed was named the 92EF population. Approximately 126 F1 individuals that appeared to have better agronomic characteristics than the Q508 of origin were selected for au-pollination. A portion of the F2 seed of these individuals was re-planned in the field. Each of the F2 plans was self-pollinated and a portion of the F3 seed was analyzed to determine the fatty acid composition. The content of oleic acid in the F3 seeds ranged from 59 to 79%). No high oleic acid was recovered in any individual (> 80%) with good characteristics and a good agronomic type. A portion of the F2 seed from the 92EF population was planted in the greenhouse to analyze the genetics of the Q508 line. The F3 seed was analyzed from the individuals of 380 F2 individuals. The levels of C18 :? of the F3 seed of the greenhouse experiment is illustrated in Figure 1. The data were tested against the hypothesis that Q508 contains two mutant genes that are semidominant and additive: the original I MC 129 mutation as well as an additional mutation. The hypothesis also assumes that the homozygotes Q508 were greater than 85% oleic acid and the homozigo Westar had oleic acid in an amount of 62 to 67%. The possible genotypes in each gene in a crossing of Q508 with Westar can be designated as: AA = Westar Fad2a BB = Weslar Fad2b aa = Q508 Fad2a "bb = Q508 Fad2b- Assuming independent segregation, we expected a ratio of phenotypes 1: 4: 6: 4: 1. The phenotypes of the heterozigo plants are assumed to be indistinguishable and therefore, the data were tested to be adapted to a 1: 14: 1 ratio of Westar homozigo: heimeric plants: homozygotes Q508.Proportion # of Phenotypes Westar alleles Genoype 1 4 AABB (Westar) 4 3 AABb, AaBB, AABb , AaBB 6 2 AaBb, AAbb, AaBb, AaBb, aaBB, AaBb 4 1 Aabb, aaBb, Aabb, aaBb 1 O aabb (Q508) Using the Chi-square analysis, the oleic acid data are adapted to a ratio of 1: 14: 1. It was concluded that Q508 differs from Westar by two main genes that are semidominant and additive and that are added independently. For comparison, the genotype of BMI 129 is aaBB. The fatty acid composition of representative F3 individuals having amounts greater than 85% oleic acid in the oil of the seeds are illustrated in Table 1 8. It is seen that the levels of saturated fatty acids have decreased in said plants, compared to the Westar.

TABLE 18 Individuals F, 92EF with > 85% C _ «- - in the Seed Oil.

EXAMPLE 6 Fatty Acid Profiles of the Leaf and Root of the Canopy Lines IMC-129. Q508 v Westar.

Plants of Q508, I MC 129 and Westar were harvested in the greenhouse.

Mature leaves, mainly expanding leaves, petioles and roots were harvested in a 6 to 8 leaf stage, frozen in liquid nitrogen and stored at a temperature of -70 ° C. the lipid extracts were analyzed by GLC as described in Example 1. The fatty acid profile data are illustrated in Table 19. The data in Table 1 9 indicate that the total leaf lipids in Q508 are higher in C? 8 content? that the content C18: 2 plus C18: 3. The opposite is true for Westar and I MC 129. The difference in the leaf lipids between Q508 and I MC 129 is consistent with the hypothesis that a second Fad2 gene is mutated to Q508. The content of C? 6: 3 in the total lipid fraction was approximately the same for all three lines, suggesting that the plastid product of the FadC gene was not affected by the Q508 mutations. To confirm that the FadC gene was not mutated, the chloroplast lipids were separated and analyzed. No changes were detected in the chloroplast of C16 fatty acids :? , C? 6: 2 or C16: 3 in the three lines. The similarity in the plastid lipids of the leaves between Q508, Wesíar and IMC 129 is consistent with the hypothesis that the second mutation in Q508 affects a microsomal Fad2 gene and not a FadC plastid gene. ro ro Ul or Oí n TABLE 19 s > EXAMPLE 7 Desaturases of Wild-type Delta-12 Fatty Acid and Mutation Sequence of B. napus. The specific primers for the FAD2 structural gene were used to clone the complete open lecíure culture (ORF) of the delta-12 desaturase D and F genes by the reverse transcriptase polymerase chain reaction (RT-PCR). RNA from the seeds of IMC 129, Q508 and Westar was isolated by standard methods and used as a template. The amplified RT fragments were used for the determination of the nucleophilic sequence. The DNA sequence of each of the genes of each line was determined by both methods of threads, by elaboration of sequences of standard dioxide strands. Sequence analysis revealed a transversion from G to A at nucleotide 316 (of the shRNA initiation codon) of gene D in both I MC 129 and Q508 compared to the Westar sequence. The transversion changes the codon in this position from GAG to AAG and results in a non-conservative substilution of glutamic acid, an acid residue, for lysine a basic residue. The presence of the same mutation in both lines was expected since the Q508 line was derived from the BMI 129. The same basic change was also detected in the Q508 and the BMI 129 when the RNA of the leaf tissue was used as a template. The mutation of G to A in nucleotide 316 was confirmed by sequencing several independent clones containing directly amplified fragments of the genomic DNA of IMC 129 and Westar. These results eliminated the possibility of a rare mutation introduced during reverse transcription and PCR in the RT-PCR protocol. It was concluded that mutant I MC 129 is due to a single basic transversion at nucleotide 316 in the coding region of the D gene of the microsomal desaturase delta 12 of rapeseed. A single basic transition from T to A was detected in nucleotide 515 of the F gene in Q508 compared to the Westar sequence. The mutation changes the codon in this position from CTC to CAC, resulting in the non-conservative substitution of a polar residue, leucine, for a polar residue, histidine in the product of the resultale gene. No mutations were found in the F gene sequence of IMC 129 compared to the sequence of the Westar F gene. These data support the conclusion that a mutation in the deslaurase gene sequence della-12 results in alterations in the fatty acid profile of the plants containing said muid gene. In addition, the data show that when a plant or species line contains two loci of desaturase della-12, the fatty acid profile of an individual having two mutated loci differs from the fatty acid profile of an individual having a mutated locus. The mutation in gene D of I MC 129 and Q508 shielded in one region has a conserved amino acid motif (His-Xaa-Xaa-Xaa-His), found in the desaturase bound to the delia-12 membrane and delinea- 15 cloned (Table 20).

TABLE 20 Alignment of the Amino Acid Sequences of the Nullified Desaturases of the Cloned Camelola Membrane.

(FadD = Delta 15 plastid, Fad3 = Delta-15 microsomal), (FadC = Delta-12 plastid, Fad2 = Delta-12 microsomal) a One letter of the amino acid code; Conservative substitutions are underlined; and the non-conservative substitutions are in bold.

EXAMPLE 8 Transcription and Translation of the Desaturases of Fatty Acid Delta-12 Microsómico. In vivo transcription was analyzed by RT-PCR analysis of stage I and stage I, developing seeds and leaf litter. The primers used to specifically amplify the F-gene RNA of the delta-12 desaturase of the indicated tissues was a 5'-GGATATGATGATGGTGAAAGA-3 'perception preparer and the 5'-TCTTTCACCATCATCATATCC-3' anti-sensing primer. The primers used to specifically amplify the D-gene RNA of the delfa-12 desaturase of the indicated liquids was the perception preparer 5'-GTTATGAAGCAAAGAAGAAAC-3 'and the anti-perception preparer 5'- GTTTCTTCTTTGCTTCATAAC-3'. The results indicated that the mRNA of both D and F genes was expressed in the seeds and tissues of the leaves of the BMI 129, Q508 and the wild-type Westar plants. In vitro transcription and translation analysis showed that a peptide of approximately 46 kD was made. That is the expected size of both the D gene products and the F gene product, based on the sum of the deduced amino acid sequence of each gene and the co-translational addition of a microsomal membrane peptide. These results rule out the possibility that mutations of non-perception or structural change are present, resulting in a product of the truncated polypeptide gene in any mutant D gene or mutant F gene. The data, in conjunction with the harms of Example 7, support the conclusion that the mullions in Q508 and BMI 129 are in the structural genes of the delta-12 fatty acid desaturase that encodes the desaturase enzymes instead of the regulatory genes.

EXAMPLE 9 Development of Gene-Specific PCR Markers. Based on a single basic change in the mutant D gene of I MC 129 described above, two 5 'PCR primers were designed. The nucleotide sequence of the primers differed only at the base (G for Westar and A for BMI 129) at the 3 'end. The primers allow the fad2 mutant and wild-type fad2 alleles to be distinguished in the DNA-based PCR assay. As there is only a single basic difference in the 5 'PCR primers, the PCR assay is very sensitive to the PCR conditions such as annealing temperature, number of cycles, amount and purity of the DNA templates used. The assay conditions have been established so as to disengage the mutant gene and the wild-type gene using the genomic DNA of I MC 129 and wild-type plants as templates. The conditions can be further optimized by varying the PCR parameters particularly with varying samples of crude DNA. A PCR assay that distinguishes the single basic mutation in I MC 129 of the wild type gene, together with the fatty acid composition analysis provides a means to simplify the segregation analysis and selection of genetic crosses comprising plants that have a mutation of the delta-12 fatty acid desaturase.

EXAMPLE 10 Transformation with Mutant Fad3 Genes and Wild Type. The Wesfar of B. napus culinae was transformed with mutant and wild-type Fad3 genes to demonstrate that the Fad3 gene for delta-1 5 desaturase, cytoplasmic linoleic desaturase of the canola is not functional. Transformation and regeneration were carried out using unarmed Agrobacterium tumefaciens following essentially the procedure described in WO 94/1 1 516. Two unarmed Agrobacteria strains were designed., each containing a Ti plasmid having the appropriate gene linked to a seed-specific promoter and a corresponding termination sequence. The first plasmid, plMC1 10, was prepared by inserting a full-length wild-type Fad3 gene into a disarmed Ti vector in sense orientation (nucleotides 208 to 1 336 of the SEQUENCE IDENTIFICATION 6 in WO 93/1 1245), flanked by a napin promoter sequence placed at 5 'for the Fad3 gene and a napin termination sequence placed at 3' for the Fad3 gene. The rapeseed napin promoter was described in patent EP 0255378. The second plasmid, pl MC205 was prepared by inserting a mutated Fad3 gene into the orientation of perception within a disassembled Ti vector. The mutant sequence contained mutations in nucleotides 41 1 and 41 3 of the chromosomal Fad3 gene described in WO93 / 1245, thereby changing the sequence for codon 96 from GAC to AAG. The amino acid in codon 96 of the gene product was changed by the amount of aspartate acid to lysine. See Table 20. A promoter fragment of bean (Faseolus vulgaris) phaseolin (7S seed storage protein) of basic pair 495 was placed in the 5 'position for the mutant Fad3 gene and a phaseolin termination sequence was placed in 3'. for the mutant Fad3 gene. The phaseolin sequence is described by Doyle et al, (1986) in J. Biol. Chem. 261, pages 9228 to 9238 and by Slightom et al, (1983) in Proc. Nati Acad. Sci. USA 80: pages 1897 to 1901. The appropriate plasmids were designed and transferred separately to the strain LBA4404 of the Agrobacteria. Each designed deformation was used to infect 5mm segments of hypocotyl explants from Wesíar seeds by means of co-culture. The infected hypocotyls were transferred to the callus medium and subsequently to the regeneration medium. Once the formed stems of the callus could be discerned, the shoots were excised and transferred to a medium of elongation. The elongated stems were cut, submerged in Rootone ™, rooted in an agar medium and transplanted to farmland to obtain fertile T1 plants. The T2 seeds were obtained by self-pollination of the resulting T1 plants. The fatty acid analysis of the T2 seeds was carried out as described above. The results are summarized in Table 21. Of the 40 transformants obtained using plasmid plMC1 10, 17 plants demonstrated wild type fatty acid profiles and 16 demonstrated overexpression. It was expected that a proportion of the transformants would show an overexpression phenotype when a functioning gene is transformed in the plants, in the orientation of the perception. Of the 307 transformed pineapples that have the pl gene MC205, none exhibited a fatty acid composition indicating overexpression. This result indicates that the product of the mutant gene fad3 is not functional, because if the production of the gene were functional, some of the transformers would have exhibited an overexpresion phenotype. ro ro Ol or Ol Ol TABLE 21 Events of Overexpression and Co-Suppression in the Westar Population Transformed with pl C205 or pl C110 OR) The fatty acid compositions of the representative transformed plants are shown in Table 22. Lines 652-09 and 663-40 are representative of plains that contain pl MC1 and exhibit an overexpression and a co-suppression phenotype, respectively. Line 205-284 is representative of the plans that comprise pIMC205 and that contain the mutant gene fad3.

TABLE 22 Fatty Acid Composition of T2 Seed from Westar Plants Transformed with PIMC205 or PIMC1 10.

EXAMPLE 1 1 Sequences of Fad2-D v Fad2-F Wild type v Mutant. The high molecular weight genomic DNA was isolated from the leaves of the pranks Q4275 (Example 5). This DNA was used as a template for the amplification of the Fad2-D and Fad2-F genes by the polymerase chain reaction (PCR). The PCR amplifications were carried out in a total volume of 100 μl with a content of 0.3 μg of genomic DNA, 200 μM of deoxyribonucleoside triphosphates, 3 mM of MgSO4, of 1 to 2 Units of DNA polymerase and 1 X regulator (supplied by a manufacturer of DNA polymerase). The cycle conditions were: 1 cycle of 1 minute at an hour of 95 ° C, followed by 30 cycles of 1 minute at a temperature of 94 ° C, 2 minutes at a temperature of 55 ° C and 3 minutes at 73 ° C C. The Fad2-D gene was amplified once using Elongase® (Gibco-BRL). The PCR preparers were: CAUCAUCAUCAUCTTCTTCGTAGGGTTCATCG and CUACUACUACUATCATAGAAGAGAAAGGTTCAG for the 5 'and 3' ends of the gene respectively. The Fad2-F gene was independently amplified 4 times, twice with Elongase® and twice with Taq polymerase (Boehringer Mannheim). The PCR preparers used were: 5 ' CAUCAUCAUCAUCATGGGTGCACGTGGAAGAA3 'and 5' CUACUACUACUATCTTTCACCATCATCATATCC3 'for the 5' and 3 'ends of the gene respectively. The amplified DNA products were resolved on an agarose gel, purified by means of JetSorb® and then annealed into pAMP1 (Gibco-BRL) by means of sequences (CAU) 4 and (CUA) 4 at the ends of the primers , and transformed into E. coli DH5a. The injections Fad2-D and Fad2-F were ordered in both threads with an automatic computer ABI PRISM 310 (Perkin-Eimer) following the manufacturer's instructions, using synthetic primers, AmpliTaq® DNA polymerase and dye finisher.

It was found that the Fad2-D gene has an intron upward current of the ATG start codon. As expected, the coding sequence of the gene contained a mutation from G to A in nucleotide 316, derived from IMC 129 (Figure 2). A single base transversion from G to A was detected at nucleotide 908 in the F sequence of the gene of Q4275 amplified products, compared to the F sequence of the wild-type gene (Figure 2). This mulation changes the codon at amino acid 303 from GGA to GAA, resulting in the non-conservative subsitiation of a glutamic acid residue by a glycine residue (Table 3 and Figure 3). Expression of the muidenle delta-12 desaturase gene of Q4275 Fad2-F in plants alters the fatty acid composition, as described earlier.

EXAMPLE 12 Rape Seed of Erucic Acid and High Oleic Acid. The culture procedure designed to produce new fatty acid compositions in rape seed is indicated in the Figure 4. In general, crosses were made between a high erucic acid line and a high oleic acid line. The elevated erucic acid line designated HEC01, (marketed under the trade name) Hero) contains approximately 45.5% erucic acid (Table 2. 3). The high oleic acid lines were designated 93GS66A-1 30 and 93GS34A-1 79 and were derived from the 93GS. See for example, Example 5 and Table 17. These lines contain approximately 84% oleic acid in their seed oil (Table 24).

TABLE 23 Fatty Acid Composition of HEC01 TABLE 24 Fatty Acid Composition of the 93GS66A-130 and 93GS34A-179 The generations F ^ of the crosses between HEC01 x 93GS66A-130 and HEC01 x 93GS34A-179 were designated 96,801 and 96,804, respectively. Fi plants 96.801 and 96.804 were self-pollinated to produce the F2 seed. In general, 622 simple seeds F2 were randomly analyzed to determine their fatty acid composition. Table 25 summarizes the average percentage and the standard deviation for the total monounsaturated content of oleic acid, eicosenoic acid and erucic acid and the total polyunsaturated and total saturated fatty acid content of these 622 seeds.

TABLE 25 The analysis of the data indicates that the frequency distributions deviate from the normal distribution. The frequency distribution of total monounsaturated long chain content is slightly skewed to the right (-0.0513), and the distribution of eicosenoic acid content is strongly skewed to the right (-1 .715). The frequency distributions of the content of oleic acid and erucic acid are strongly tilted to the left (0.397 and 0.177 respectively). The inclination was calculated using a Lotus 1 -2-3 for Windows (version 5.0). Table 26 describes the characteristics of the selected populations within the total seed population. For example, 1 51 seeds had a long chain monounsaturated fatty acid content greater than 82% (Table 26, column B). Within this population the average content of oleic, eicosenoic and erucic acid was approximately 48%, 16% and 1%, respectively. The total content of polyunsaturated fatty acid (Cl.sub.8: 2, Cl.sub.8: 3, and C20: 2) was approximately 9%, and the total content of saturated fatty acid was less than 7%. Forty-seven of the 622 seeds had a long-chain monounsaturated content greater than 85% (Table 26, column C). The average content of oleic, eicosenoic and erucic acid within these seeds was 51%, 17% and 17% respectively. The saturated fatty acids total and polyunsaturated were each less than 7%.

Twenty-three of these seeds had an eicosenoic acid content greater than 19% > (Table 26, column F). Within these seeds, the average content of oleic acid and erucic acid was approximately 44% and 19% respectively. The total polyunsaturated fatty acids were less than 10%), and the total saturated fatty acids were less than 7%. ro ro Ül o 01 Ol TABLE 26 A = > 80% total long chain monounsaturated content, n = 247; B = > 82% total long chain monounsaturated content, n = 151; C = > 85% total long chain monounsaturated content, n = 47; D = > 15% erucic acid, n = 318; E = > 15% ecosenoic acid, n = 33; F = > 19% ecosenoic acid, n = 23 The fatty acid composition of the selected single seeds is presented in Table 27. V800655.334 was a single seed having a long chain monounsaturated fatty acid content of about 84%. The conlicide of oleic acid, eicosanoic acid and erucic acid was 33.48% > , 7.14% and 32.23% respectively. The total content of polyunsaturated fatty acid was about 10%. The content of llnoleic, a-linolenic and erucic acid was 3.54%, 6.01% and 0.15%) respectively. V800655.126 was a simple seed having a long chain monounsaturated fatty acid content of approximately 85% (42.67% oleic acid, 16.21% eicosenoic acid, and 25.37% erucic acid). The ionic content of polyunsaturated fatty acid was about 8% (4.87% llnoleic acid, 3.05% a-linolenic acid, and 0.13%> eicosadienoic acid). V800654.9 was a single seed having a long chain monounsaturated fatty acid content of 89% (51.53% oleic acid, 16.94%) of eicosenoic acid, and 19.24% of erucic acid). The total content of polyunsaturated fatty acid was approximately 8% >; (4.87% linoleic acid, 3.05% a-linolenic acid, and 0.13% eicosadienoic acid). Simple seeds having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% were planted in a greenhouse, allowed to grow to maturity and were self-pollinated . The seeds of each plant (generation F3) were harvested. A sample of bulk seeds from each of the F2 plants was analyzed to determine the fatty acid composition.

TABLE 27 Fatty Acid Composition of Selected Single Seeds.

EXAMPLE 13. Additional crossings were made between Hero and several lines of high oleic acid (Table 28) to increase the erucic acid content of the seed through a reduction in the content of the polyunsaturated and increase the total monounsaturated content. The high oleic acid lines included 048X058 and Q4275X663-40. Line 048X058 resulted from a crossing of two separate transformed lines. Line 048X058 contains a co-suppression event resulting from the introduction of transgene 663-40 described above, and a second event of co-suppression resulting from a transgene including an oleosin promoter linked to an oleic desaturase gene. The Q4275X663-40 line was derived from a crossing of Q4275 (Example 5 and Table 17) by 663-40. Line 663-40 contains a co-suppression event resulting from a transgene that includes a napin promoter linked to a lignole desaturase gene. The plañías of each of the lines were culíivadas in chambers of culture in a regime of 16 hours of light and temperature day / night of 23/17 ° C. The flowers were castrated before opening and covered to avoid cross-pollination. The next day, the stigmas of the castrated flowers were polinated with the desired pollen donor. At the maturation of the pod, the F1 seed was harvested. ro ro I heard or Ol Ol TABLE 28 Block of Crosses 00 The F1 seed generated from the crosses in Table 28 was advanced to the generation of F2 seeds in the growth chamber.

Ten seeds from each crossing were planted individually. At flowering the plants were covered with bags to ensure self-pollination. The F2 seeds were harvested at maturity. The seeds were germinated on filter paper at room temperature in the dark. Eighteen to 24 hours after the start of germination, a cotyledon was removed from the seed for the extraction of the fatty acids. The fatty acid composition was demineralized using gas chromatography. The selected half seeds F2 have a high erucic acid content, which is illustrated in Tables 29 and 30.

The half F2 seeds were planted in the soil and harvested under the conditions of a growth chamber described above. At flowering the plants were covered with bags for auopolopolization. After maturation, the F3 seed was harvested and its fatty acid composition was analyzed. The seeds were analyzed using a sample size of 10 to 15 seeds. The results of the analysis will be found in Tables 31 and 32. ro ro Ül or í Ol I? BL? _2_. Fatty Acid Composition of HEHOA Seeds [HE101X (048X058)] oo? ro ro Ol or Ol Ül TABLE 30 Fatty Acid Composition of Stocks HEHOC Seeds [HE101X (Q4275X663-40 =] 00 ro ro Ül or Ol Ol TABLE 31 Fatty Acid Composition of F3 Lines of 97HEHOA [HE101X (048X052)] 00 Ol ro ro Ol or Ol Ül IABLA_32 Fatty Acid Composition of HEHOC F3 Lines [HE101X (Q4275X663-40)] 00 The seeds F3 HEHOC-214 were planted and cultivated under the conditions of a growth chamber and self-pollinators as described above. Upon maturation, the F4 seeds were harvested and their fatty acid composition analyzed, ullecting bulk samples of 10 to 15 seeds. The fatty acid composition of the seed of the three F4 lines is illustrated in Table 33. All three samples had a long chain monounsaturated fatty acid content greater than 82% and an erucic acid content greater than 37% based on the total fatty acid composition. The genes that affect the fatty acid composition are still segregating in this material from generation F. The selection in subsequent generations will determine the genetic composition and will result in lines having a fatty acid composition of the seed of about 25 to 30% oleic acid, of about 3 to 4% linoleic acid, of about 1. to 2% a-linolenic acid, from about 45 to 50% erucic acid and from about 10 to 13% eicosenoic acid. ro ro Ol or Ol Ol I? BLAÜS Fatty Acid Composition of F4 Lines of HEHOC 214 00 00 EXAMPLE 14 Seeds from the lines in Table 31 were plotted on two separate lines. The plants of each line were allowed to polinate open. The oil was extracted from the seeds produced in each line; the fatty acid compositions of each oil are illustrated in Table 34. A sample of each oil was refined and bleached (row 1 monounsaturated long chain high) or refined, bleached and deodorized (monounsaturated long chain high row 2). The oil in row 1 had a saturated total content of 5.55%, a long chain monounsaturated fatty acid content of 85.23% and a total content of polyunsaturated fatty acid of 6.80%. The oil in row 2 had a total saturated fatty acid content of 5.22%, a long-chain monounsaturated fatty acid content of 82.90% and a lowered content of polyunsaturated fatty acid of 9.56%. The iodine values of the oil in rows 1 and 2 were 79 and 81.7, respectively. The oil in row 1, which was not deodorized, contained 420 ppm of tocopherol. The oil in row 2 contained 280 ppm tocopherol. The oils in rows 1 and 2 had average oxidative stabilities of 70 AOM hours (n = 2, 69 and 71 AOM hours) and 49.5 AOM hours (n = 2, 48 and 51 AOM hours), respectively.

TABLE 34 Fatty Acid Composition of Monounsaturated Rape Seed Oil of High Long Chain.

Table 35 provides characteristics of the oils in row 1 and 2, which were determined by differential scanning calorimetry using a Perkin Elmer Model 7 differential scanning calorimeter. Samples of 7 to 12 milligrams were placed in the sample trays; sealed and loaded in the self-soler. The samples were cooled to an initial temperature of 20 ° C which was maintained for 1 minute, at a temperature of -30 ° C in a range of 40 ° C per minute. The samples were kept at a temperature of -30 ° C for 10 minutes, then heated to 75 ° C in a range of 5 ° C per minute to obtain a melting curve. The samples were held at 75 ° C for 10 minutes and then cooled to -30 ° C in a range of 5 ° C per minute to obtain a cooling curve. The results show that these oils, which have long chain monounsaturated content greater than 82% and an erucic acid condense greater than 1 5%, exhibit melting points of about 2-3 ° C. For comparison, trierucin is a solid at room temperature.

TABLE 35 Characteristics of Monounsaturated Rape Seed Oil of High Long Chain.

EXAMPLE 15 Brassica napus seeds of the IMC 129 variety were mutagenized with MNNG as described in Example 5. The treated seeds were cultured as described in Example 5 and a selection was made by the stearic acid. decreased of the seed, or the content of palmitic acid decreased in an M3 generation. The plants of the two selected lines, designated ZW1441 (diminished palmitic acid) and Y30137 (diminished stearic acid), were crossed with HE101. Descendants ZW1441 XHE101 were selected that produced seeds that have reduced palmitic acid and high erucic acid. Y301 37XHE101 descendants were selected that produced seeds having decreased spheric acid and high erucic acid. The fatty acid composition of the seeds of the selected lines representative of generation F are illustrated in Table 36. The results show that seeds having a long chain monounsaturated content of about 82% or greater can be achieved, a conenido of erucic acid of 15% > or greater and a legal content of less than 4% (for example, from approximately 2.0 to approximately 4.0%). ro ro Ül or Ol Ol TABLE 36 Fatty Acid Composition of Lines F4 CD To a point not yet indicated, it will be understood by those skilled in the art that any of the different specific embodiments described herein may be further modified to incorporate features that are shown in other specific embodiments. All patents, publications and other references herein are incorporated by reference in their entirety. The above detailed description has been provided only for a better understanding of the invention and should not be understood as an unnecessary limitation thereof, since those skilled in the art will appreciate some modifications without departing from the spirit and scope of the appended claims.

LIST OF SEQUENCES < 110 > Kodali, Dharma R. Fan, Zhegong DeBopte, Lorin R. < 120 > PLANTS, SEEDS AND OILS WHICH HAVE A TOTAL, ELEVATED MONOSATURATED FATTY ACID CONTENT < 130 > 07148 / 097WO1 < 150 > 09 / 128,602 < 151 > 1998-08-03 < 160 > 18 < 170 > FastSEQ for Windows Version 3.0 < 210 > 1 < 211 > 1155 < 212 > DNA < 213 > Brassica napus < 220 > < Z21 > CDS < 222 > (1) ... (1152) < »21 > doubtful < 222 > (133) ... (133) < 223 > Xaa = * Pro or Leu < 221 > doubtful < 222 > (194) ... (194) < 223 > Xaa - Leu 221 > doubtful < 222 > (246) ... (246) < 223 > Xaa - He or Val J2l > doubtful < 222 > (262) ... (262) < 223 > Xaa = He or Val < £ 21 > doubtful < 222 > (345) ... (345) < 223 > Xaa - Tyr < 400 > 1 atg ggt gca ggt gga aga atg cag gtg tet ect ecc tec aaa aag tet 48 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 gaa acc gac aac ate aag cgc gta ecc tgc gag here ceg ecc ttc act 96 Glu Thr Asp Asn He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 gtc gga gaa etc aag aaa gca ate cea ceg cae tgt ttc aaa cgc teg 144 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 ate ect cgc tet ttc tec tac etc ate tgg gac ate ate ata gee tec 192 He Pro Arg Ser Phe Ser Tyr Leu He Tip Asp He He He Wing Be Ser 50 55 60 tgc ttc tac tac gtc gee acc act tac ttc ect etc etc ect drops ect 240 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 etc tec tac ttc gee tgg ect etc tac tgg geg tgc cag ggc tgc gtc 288 Leu Ser Tyr Phe Wing Trp Pro Leu Tyr Trp Wing Cys Gln Gly Cys Val 85 90 95 cta acc ggc gtc tgg gtc ata gee falls gag tgc ggc falls falls gee ttc 336 Leu Thr Gly Val Trp Val He Wing His Glu Cys Gly His His Ala Phe 100 105 110 age gac tac cag tgg ctg gac gac acc gtc ggc etc ate ttc falls tec 384 Ser Asp Tyr Gln Trp Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 ttc etc etc gtc cyt tac ttc tec tgg aag tac agt cat cga cgc cae 432 Phe Leu Leu Val Xaa Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His 130 135 140 cat toe aac act ggc tec etc gag aga gac gtg ttt gtc ecc aag 480 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 aag aag ate gac ate aag tg tac ggc aag tac etc aac act ect ttg 528 Lys Lys Ser Asp He Lys Trp Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 gga cgc acc gtg atg tta acg gtt cag ttc act etc ggc tgg ect ttg 576 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Trp Pro Leu 180 185 190 tac ttr gcc ttc aac gtc tcg ggg aga ect tac gac ggc ggc ttc gct 624 Tyr Xaa Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 tgc cat ttc falls ecc aac gct ecc ate tac aac gac cgt gag cgt etc 672 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 cag ata tac ate tec gac gct ggc ate etc gcc gtc tgc tac ggt etc 720 Gln He Tyr He Ser Asp Wing Gly pe Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 tac cgc tac gct gct rtc ca g gt gtt gcc tcg atg gtc tgc ttc tac 768 Tyr Arg Tyr Ala Ala Xaa Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 gga gtt ect ctt ctg rtt gtc aac ggg ttc tta gtt ttg ate act tac 816 Gly Val Pro Leu Leu Xaa Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 ttg cag falls acg cat ect tec ctg ect falls tat gac tcg tet gag tgg 864 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Trp 275 280 285 gat tgg ttg agg gga gct ttg gcc acc gtt gac aga gac tac gga ate 912 Asp Trp Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly He 290 295 300 ttg aac aag gtc ttc falls aat ate acg gac acg falls gtg gcg cat falls 960 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 ctg ttc tcg acc atg ceg cat tat cat gcg atg gaa gct acg aag gcg 1008 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Wing Thr Lys Wing 325 330 335 ata aag ceg ata ctg gga gag tat tay cag ttc gat ggg acg ceg gtg 1056 He Lys Pro He Leu Gly Glu Tyr Xaa Gln Phe Asp Gly Thr Pro Val 340 345 350 gtt aag gcg atg tgg agg gag gcg aag gag tgt ate tat gtg gaa ceg 1104 Val Lys Ala Met Trp Arg Glu Wing Lys Glu Cys He Tyr Val Glu Pro 355 360 365 gac agg caa ggt gag aag aaa ggt gtg ttc tgg tac aac aat aag tta 1152 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Trp Tyr Asn Asn Lys Leu 370 375 380 tga 1155 < 210 > 2 < 211 > 384 < 212 > PRT < 213 > Brassica napus < 220 > < 221 > DOUBTFUL < 222 > (133) ... (133) < 223 > Xaa = Pro or Leu < 221 > DOUBTFUL < 222 > (194) ... (194) < 223 > Xaa = Leu < 221 > DOUBTFUL < 222 > (246) ... (246) < 223 > Xaa - He or Val < 221 > DOUBTFUL < 222 > (262) ... (262) < 223 > Xaa = He or Val < 221 > DOUBTFUL 222 > (345) ... (345) < 223 > Xaa - Tyr < 400 > 2 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 Glu Thr Asp Asn He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 He Pro Arg Ser Phe Ser Tyr Leu lie Trp Asp He He He Wing Be 50 55 60 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 Leu Ser Tyr Phe Wing Trp Pro Leu Tyr Trp Wing Cys Gln Gly Cys Val 85 90 95 Leu Thr Gly Val Trp Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 Ser Asp Tyr Gln Trp Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 Phe Leu Leu Val Xaa Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 Lys Lys Ser Asp pe Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 Tyr Xaa Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Ala 195 200 205 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 Gln He Tyr lie Ser Asp Ala Gly He Leu Ala Val Cys Tyr Gly Leu 225 230 235 240 Tyr Arg Tyr Ala Ala Xaa Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 Gly Val Pro Leu Leu Xaa Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 Asp Tf Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly He 290 295 300 Leu Asn Lys Val Phe His Asn lie Thr Asp Thr His Val Wing His His 305 310 315 320 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Ala Thr Lys Ala 325 330 335 He Lys Pro lie Leu Gly Glu Tyr Xaa Gln Phe Asp Gly Thr Pro Val 340 345 350 Val Lys Ala Met Tf Arg Glu Ala Lys Glu Cys He Tyr Val Glu Pro 355 360 365 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 < 210 > 3 < 211 > 1155 212 > DNA < 213 > Brassica napus < 220 > < 221 > CDS < 222 > (1) ... (1152) < 223 > < 221 > doubtful 222 > (133) ... (133) 223 > Xaa «= Pro or Leu 22l > doubtful < 222 > (194) ... (194) < 223 > Xaa = Leu < 221 > doubtful < 222 > (246) ... (246) < 223 > Xaa - He or Val < 221 > doubtful < 222 > (262) ... (262) < 223 > Xaa = He or Val < 221 > doubtful 222 > (345) ... (345) < 223 > Xaa = Tyr < 400 > 3 ag ggt gca ggt gga aga atg cag gtg tet ect ecc tec aaa aag tet 48 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 gaa acc gac aac ate aag cgc gta ecc tgc gag here ceg ecc ttc act 96 Glu Thr Asp Asn He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 gtc gga gaa etc aag aaa gca ate cea ceg cae tgt ttc aaa cgc tcg 144 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 ate ect cgc tet ttc tec tac etc ate tgg gac ate ate ata gcc tec 192 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Be Ser 50 55 60 tgc ttc tac tac gtc gcc ac act tac ttc ect etc etc ect drops ect 240 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 etc tec tac tcc gcc tgg ect etc tac tgg gcc tgc cag ggc tgc gtc 288 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 cta acc ggc gtc tgg gtc ata gcc falls aag tgc ggc falls drops gcc ttc 336 Leu Thr Gly Val Tf Val He Wing His Lys Cys Gly His His Wing Phe 100 105 110 figc gac tac cag tgg ctg gac gac acc gtc ggc etc ate ttc falls toe 384 Be Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 ttc etc etc gtc cyt tac ttc tec tgg aag tac agt cat cga cgc drops 432 Phe Leu Leu Val Xaa Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 cat toe aac act ggc toe etc gag aga gac gaa gtg ttt gtc ecc aag 480 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 aaag aag ate gac ate aag tg tac ggc aag tac etc aac aac ect ttg 528 Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 gga cgc acc gtg atg tta acg gtt cag ttc act etc ggc tgg ect ttg 576 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 tac ttr gcc ttc aac gtc tcg ggg aga ect tac gac ggc ggc tcc gct 624 Tyr Xaa Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 tgc cat ttc drops ecc aac gct ecc ate tac aac gac cgt gag cgt etc 672 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 cag ata tac ato toe gac gct ggc ato etc gcc gtc tgc tac ggt etc 720 Gln He Tyr He Ser Asp Wing Gly He Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 tac cgc tac gct gct rtc ca g gt gtt gcc tcg atg gtc tgc ttc tac 768 Tyr Arg Tyr Ala Ala Xaa Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 gga gtt ect ctt ctg rtt gtc aac ggg ttc tta gtt ttg ate act tac 816 Gly Val Pro Leu Leu Xaa Val Asn Gly Phe Leu Val Leu lie Thr Tyr 260 265 270 ttg cag falls acg cat ect toe ctg ect falls tat gac tcg tot gag tgg 864 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 gat tgg ttg agg gga gct ttg gcc acc gtt gac aga gac tac gga ate 912 Asp Tf Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly lie 290 295 300 ttg aac aag gtc ttc falls aat ato acg gac acg falls gtg gcg cat falls 960 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 etg ttc tcg acc atg ceg cat tat cat gcg atg gaa gct acg aag gcg 1008 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Wing Thr Lys Wing 325 330 335 ata aag ceg ata ctg gga gag tat tay cag ttc gat ggg acg ceg gtg 1056 He Lys Pro He Leu Gly Glu Tyr Xaa Gln Phe Asp Gly Thr Pro Val 340 345 350 gtg aag gcg atg tgg agg gag gcg aag gag tgt ate tat gtg gaa ceg 1104 Val Lys Ala Met Tf Arg Glu Wing Lys Glu Cys He Tyr Val Glu Pro 355 360 365 gac agg caa ggt gag aag aaa ggt gtg tte tgg tac aac aat aag tta 1152 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 tga 1155 < 210 > 4 < 211 > 384 < 212 > PRT < 213 > Brassica napus < 220 > < 221 > DOUBTFUL < 222 > (133) ... (133) < 223 > Xaa = Pro or Leu < 221 > DOUBTFUL < 222 > (194) ... (194) < 223 > Xaa = Leu < 221 > DOUBTFUL 222 > (246) ... (246) < 223 > Xaa = lie or Val < 221 > DOUBTFUL < 222 > (262) ... (262) < 223 > Xaa = pe or Val 221 > DOUBTFUL 222 > (345) ... (345) < 223 > Xaa = Tyr < 400 > 4 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 Glu Thr Asp Asn He Lys Arg Val Pro Cys Glu Tbx Pro Pro Phe Thr 20 25 30 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 40 45 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Being 50 55 60 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 Leu Thr Gly Val Tf Val He Wing His Lys Cys Gly His His Wing Phe 100 105 110 Being Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 Phe Leu Leu Val Xaa Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 Tyr Xaa Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Ala 195 200 205 Cys His Phe His Pro Asn Ala Pro lie Tyr Asn Asp Arg Glu Arg Leu 210 215 220 Gln He Tyr He Ser Asp Ala Gly He Leu Ala Val Cys Tyr Gly Leu 225 230 235 240 Tyr Arg Tyr Ala Ala Xaa Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 Gly Val Pro Leu Leu Xaa Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 Asp Tf Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly He 290 295 300 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Ala Thr Lys Ala 325 330 335 He Lys Pro He Leu Gly Glu Tyr Xaa Gln Phe Asp Gly Thr Pro Val 340 345 350 Val Lys Wing Met Tf Arg Glu Wing Lys Glu Cys He Tyr Val Glu Pro 355 360 365 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 210 > 5 211 > 1155 < 212 > DNA < 213 > Brassica napus < 220 > < 221 > CDS 222 (1) ... (1152) < 400 > 5 atg ggt gca ggt gga aga atg cag gtg tet ect ecc tec aag aag tet 48 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 gaa acc gac acc ate aag cgc gta ecc tgc gag here ceg ecc ttc act 96 Glu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 gtc gga gaa etc aag aaa ce ce cae cae tgt ttc aaa cgc tcg 144 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 ato ect cgc tet ttc toe tac etc ato tgg gac ato ato gcc toe 192 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Be 50 55 60 tgc tte tac tac gtc gcc acc act tac tct ect cte cte ect cae ect 240 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Pro His Pro 65 70 75 80 etc tec tac ttc gcc tgg ect cte tac tgg gcc tgc ca ggg tgc gtc 288 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 cta acc ggc gtc tgg gtc ata gcc falls gag tgc ggc falls falls gcc ttc 336 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 age gac tac cag tgg ctt gac gac acc gtc ggt etc ate ttc falls toe 384 As Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 ttc etc etc gtc ect tac ttc toe tgg aag tac agt cat cga cgc falls 432 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 cat tec aac act ggc toe cte gag aga gac gaa gtg ttt gto ecc aag 480 His SCT Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 aag aag aga aa tg aa tgg taag g aac aac act ect tct 528 Lys Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 gga cgc acc gtg atg tta acg gtt cag ttc act etc ggc tgg ceg ttg 576 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 tac tta gcc ttc aac gtc tcg gga aga ect tac gac ggc ggc tto gct 624 Tyr Leu Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 tgc cat ttc falls ecc aac gct ecc ato tac aac gac cgc gag cgt etc 672 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 cag ata tac ate toe gac gct ggc ato cte gcc gtc tgc tac ggt etc 720 Gln? Le Tyr He Ser Asp Wing Gly He Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 tto cgt tac gcc gcg cag gga gtg gcc tog atg gtc tgc tte tac 768 Phe Arg Tyr Ala Ala Ala Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 gga gto ceg ctt ctg gt aat ggt ttc gtg gtg ttg ate act tac 816 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 ttg cag falls acg cat ect tec ctg ect falls tac gat teg tec gag tgg 864 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 gat tgg ttg agg gga gct ttg gct acc gtt gac aga gac tac gga ate 912 Asp Tf Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly He 290 295 300 ttg aac aag gtc tto falls aat att acc gac acg falls gtg gcg cat cat 960 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 ctg ttc toe acg atg ceg cat tat drops gcg atg gaa gct acc aag gcg 1008 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Wing Thr Lys Wing 325 330 335 ata aag ceg ata ctg gga gag tat tat cag ttc gat ggg acg ceg gtg 1056 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 gtt aag gcg atg tgg agg gag gcg aag gag tgt ate tat gtg gaa ceg 1104 Val Lys Ala Met Tf Arg Glu Wing Lys Glu Cys He Tyr Val Glu Pro 355 360 365 gac agg caa ggt gag aag aaa ggt gtg ttc tgg tac aac aat aag tta 1152 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 tga 1155 < 210 > 6 < 211 > 384 < 212 > PRT < 213 > Brassica napus < 400 > 6 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 10 15 Glu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 Val Gly Glu Leu Lys Lys Wing Pro Pro His Cys Phe Lys Arg Ser 35 40 45 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Ser 50 55 60 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 Leu Ser Tyr Phe Ala Tf Pro Leu Tyr Tf Ala Cys GIn Gly Cys Val 85 90 95 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 Ser Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Are His 130 135 140 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lvs 145 150 155 160 Lys Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 Tyr Leu Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 Gln He Tyr Wing Being Asp Wing Gly He Leu Wing Val Cys Tyr Glv Leu 225 230 235 240 Phe Arg Tyr Wing Wing Wing Gln Gly Val Wing Wing Met Val Cys Phe Tvr 245 250 255 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 Leu GLa His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 Asp Tf Leu Arg Gly Wing Leu Wing Thr Val Asp Arg Asp Tyr Gly He 290 295 300 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Ala Thr Lys Ala 325 330 335 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 Val Lys Wing Met Tf Arg Glu Wing Lys Glu Cys lie Tyr Val Glu Pro 355 360 365 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 < 210 > 7 < 211 > 1155 < 212 > DNA < 213 > Brassica napus < 220 > < 221 > CDS < 222 > (1) ... (1152) 400 > 7 atg ggt gca ggt gga aga atg cag gtg tet ect ecc toe aag aag tot 48 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 gaa acc gac acc ato aag cgc gta ecc tgc gag ac ceg ecc tto act 96 Olu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 gto gga gaa etc aag aaa ce ce cae cae tgt ttc aaa cgc tcg 144 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 ate ect cgc tet tic tec tac etc ate tgg gac ato ate ata gcc toe 192 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Be Ser 50 55 60 tgc tte tac tac gtc gcc ac act tac ttc ect etc etc ect drops ect 240 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 etc toe tac tcc gcc tgg ect etc tac tgg gcc tgc ca ggg tgc gtc 288 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 cta acc ggc gtc tgg gtc ata gcc falls gag tgc ggc falls falls gcc ttc 336 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Ala Phe 100 105 110 age gac tac cag tgg ctt gac gac acc gto ggt etc ato ttc falls toe 384 Ser Asp Tyr GIn Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 ttc etc etc gtc ect tac ttc toe tgg aag tac agt cat cga cgc cae 432 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 cat toe aac act ggc toe etc gag aga gac ga gtg ttt gto ecc aag 480 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 aaag aaag aa aa aa aa aa aa aa aa aa aa aa aa aa e a t a t a t t a t t a t t a t tg 528 Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr His Asn Asn Pro Leu 165 170 175 gga cgc acc gtg atg tta acg gtt cag tto act etc ggc tgg ceg ttg 576 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 tac tta gcc ttc aac gtc tcg gga aga ect tac gac ggc ggc tcc gct 624 Tyr Leu Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 tgc cat ttc drops ecc aac gct ecc ato tac aac gac cgc gag cgt etc 672 Cys His Phe His Pro Asn Ala Pro pe Tyr Asn Asp Arg Glu Arg Leu 210 215 220 cag ata tac ate toe gac gct ggc ato cte gcc gto tgc tac ggt cte 720 Gln He Tyr He Ser Asp Wing Gly He Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 ttc cgt tac gcc gcc gcg cag gga gtg gcc tcg atg gtc tgc ttc tac 768 Phe Arg Tyr Ala Ala Ala Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 gga gto ceg ctt ctg att gto aat ggt ttc etc gtg ttg ato act tac 816 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 ttg cag falls acg cat ect toe ctg ect falls tac gat tcg tec gag tgg 864 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 gat tgg ttg agg gga gct ttg gct acc gtt gac aga gac tac gga ate 912 Asp Tf Leu Arg Gly Wing Leu Wing Thr Val Asp Arg Asp Tyr Gly He 290 295 300 ttg aac aag gtc ttc falls aat att acc gac acg falls gtg gcg cat cat 960 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 ctg ttc toe acg atg ceg cat tat drops gcg atg gaa gct acc aag gcg 1008 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Wing Thr Lys Wing 325 330 335 ata aag ceg ata ctg gga gag tat tat cag ttc gat ggg acg ceg gtg 1056 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 gtt aag gcg atg tgg agg gag gcg aag gag tgt ate tat gtg gaa ceg 1104 Val Lys Ala Met Tf Arg Glu Wing Lys Glu Cys He Tyr Val Glu Pro 355 360 365 gac agg cag ggt gag aag aaa ggt gtg tte tgg tac aac aat aag tta 1152 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 tga 1155 < 210 > 8 < 211 > 384 < 212 PRT < 213 > Brassica napus < 400 > 8 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 Glu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Be 50 55 60 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Pro His Pro 65 70 75 80 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 Ser Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 Lys Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr His Asn Asn Pro Leu 165 170 175 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 Tyr Leu Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 Gln He Tyr He Ser Asp Wing Gly He Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 Phe Arg Tyr Wing Wing Wing Gln Gly Val Wing Wing Met Val Cys Phe Tyr 245 250 255 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Glu Tf 275 280 285 Asp Tf Leu Arg Gly Wing Leu Wing Thr Val Asp Arg Asp Tyr Gly He 290 295 300 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Ala Thr Lys Ala 325 330 335 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 Val Lys Ala Met Tf Arg Glu Ala Lys Glu Cys He Tyr Val Glu Pro 355 360 365 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 210 > 9 211 > 1155 < 212 > DNA < 213 > Brassica napus < 220 > < 221 > CDS < 222 > (!) ... (! 152) < 400 > 9 atg ggt gca ggt gga aga atg ca gtg tot ect ecc toe aag aag tot 48 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 10 15 gaa acc gac acc atoc a cgc gta ecc tgc gag here ceg ecc ttc act 96 Glu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 20 25 30 gtc gga gaa etc aag aaa gca ato cea ceg cae tgt ttc aaa cgc tcg 144 Val Gly Glu Leu Lys Lys Ala He Pro Pro His Cys Phe Lys Arg Ser 35 40 45 ato ect cgc tet tto toe tac etc ato tgg gac ato ate ata gcc toe 192 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Be 50 55 60 tgc tto tac tac gto gcc acc act tac tto ect etc etc ect drops ect 240 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 cte toe tac ttc gcc tgg ect cte tac tgg gcc tgc ca ggg tgc gtc 288 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 cta acc ggc gtc tgg gtc ata gcc falls gag tgc ggc falls falls gcc ttc 336 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 age gac tac cag tgg ctt gac gac acc gto ggt etc ato tto falls toe 384 Being Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 ttc etc etc gtc ect tac ttc tec tgg aag tac agt cat cga cgc falls 432 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 cat toe aac act ggc toe etc gag aga gac gaa gtg ttt gtc ecc aag 480 His SCT Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 aaag aag aag aag aag tag taag gag aag tac etc aac aac ect ttg 528 Lys Lys Ser Asp lie Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 gga cgc acc gtg atg tta acg gtt cag tte act etc ggc tgg ceg ttg 576 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 tac tta gcc ttc aac gto tog gga aga ect tac gac ggc ggc ttc gct 624 Tyr Leu Wing Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Wing 195 200 205 tgc cat tto drops ecc aac gct ecc ato tac aac gac cgc gag cgt etc 672 Cys His Phe His Pro Asn Ala Pro He Tyr Asn Asp Arg Glu Arg Leu 210 215 220 cag ata tac ate tec gac gct ggc ato etc gcc gto tgc tac ggt etc 720 Gln He Tyr He Ser Asp Wing Gly He Leu Wing Val Cys Tyr Gly Leu 225 230 235 240 tte cgt tac gcc gcc gcg cag gga gtg gcc tog atg gte tgc ttc tac 768 Phe Arg Tyr Ala Ala Ala Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 gga gto ceg ctt ctg att gtc aat ggt ttc etc gtg ttg ato act tac 816 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 ttg cag falls acg cat ect toe ctg ect falls tac gat tog tog gag tgg 864 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu T 275 280 285 gat tgg ttg agg gga gct ttg gct acc gtt gac aga gac tac gaa ato 912 Asp Tf Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Glu He 290 295 300 ttg aac aag gtc ttc falls aat att acc gac acg falls gtg gcg cat cat 960 Leu Asn Lys Val Phe His Asn He Thr Asp Thr His Val Wing His His 305 310 315 320 ctg tto toe acg atg ceg cat tat falls gcg atg gaa gct acc aag gcg 1008 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Wing Thr Lys Wing 325 330 335 ata aag ceg ata ctg gga gag tat tat cag tto gat ggg acg ceg gtg 1056 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 gtt aa gcg atg tgg agg gag gcg aag gag tgt ato tat gtg gaa ceg 1104 Val Lys Ala Met Tf Arg Glu Ala Lys Glu Cys He Tyr Val Glu Pro 355 360 365 gac agg caa ggt gag aag aaa ggt gtg tto tgg tac aac aat aag tta 1152 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 tga 1155 < 210 > 10 < 211 > 384 < 212 > PRT < 213 > Brassica napus 400 > 10 Met Gly Wing Gly Gly Arg Met Gln Val Ser Pro Pro Ser Lys Lys Ser 1 5 10 15 Glu Thr Asp Thr He Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 25 30 Val Gly Glu Leu Lys Lys Wing Pro Pro His Cys Phe Lys Arg Ser 40 45 He Pro Arg Ser Phe Ser Tyr Leu He Tf Asp He He He Wing Being 50 55 60 Cys Phe Tyr Tyr Val Wing Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 65 70 75 80 Leu Ser Tyr Phe Wing Tf Pro Leu Tyr Tf Wing Cys Gln Gly Cys Val 85 90 95 Leu Thr Gly Val Tf Val He Wing His Glu Cys Gly His His Wing Phe 100 105 110 Being Asp Tyr Gln Tf Leu Asp Asp Thr Val Gly Leu He Phe His Ser 115 120 125 Phe Leu Leu Val Pro Tyr Phe Ser Tf Lys Tyr Ser His Arg Arg His 130 135 140 His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 145 150 155 160 Lys Ser Asp He Lys Tf Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 165 170 175 Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Tf Pro Leu 180 185 190 Tyr Leu Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Gly Phe Ala 195 200 205 Cys His Phe His Pro Asn Ala Pro lie Tyr Asn Asp Arg Glu Arg Leu 210 215 220 Gln lie Tyr pe Ser Asp Ala Gly He Leu Ala Val Cys Tyr Gly Leu 225 230 235 240 Phe Arg Tyr Ala Ala Ala Gln Gly Val Ala Ser Met Val Cys Phe Tyr 245 250 255 Gly Val Pro Leu Leu He Val Asn Gly Phe Leu Val Leu He Thr Tyr 260 265 270 Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Tf 275 280 285 Asp Tf Leu Arg Gly Wing Leu Wing Thr Val Asp Arg Asp Tyr Glu He 290 295 300 Leu Asn Lys Val Phe His Asn lie Thr Asp Thr His Val Wing His His 305 310 315 320 Leu Phe Ser Thr Met Pro His Tyr His Wing Met Glu Ala Thr Lys Ala 325 330 335 He Lys Pro He Leu Gly Glu Tyr Tyr Gln Phe Asp Gly Thr Pro Val 340 345 350 Val Lys Ala Met Tf Arg Glu Ala Lys Glu Cys He Tyr Val Glu Pro 355 360 365 Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Tf Tyr Asn Asn Lys Leu 370 375 380 210 > 11 < 211 > 21 < 212 > DNA < 213 > Other nucleic acid < 400 > 11 ggatatgatg atggtgaaag to 21 210 > 12 < 211 > 21 < 212 > DNA < 213 > Other nucleic acid < 400 > 12 tottteacca tcatcatatc c 21 < 210 > 13 < 211 > 21 212 > DNA < 213 > Other nucleic acid < 400 > 13 gttatgaagc aaagaagaaa c 21 < 210 > 14 211 > 21 < 212 > DNA < 213 > Other nucleic acid < 400 > 14 gtttcttctt tgcttoataa c 21 < 210 > 15 < 211 > 32 < 212 > DNA < 213 > Other nucleic acid < 400 > 15 caucaucauc aucttcttog tagggttcat cg 32 < 210 > 16 < 211 > 33 212 > DNA < 213 > Other nucleic acid < 400 > 16 cuacuacuac uatcatagaa gagaaaggtt cag 33 210 > 17 211 > 32 212 > DNA < 213 > Other < 400 > 17 caucaucauc aucatgggtg cacgtggaag aa 32 < 210 > 18 211 > 33 212 > DNA < 213 > Other < 400 > 18 cuacuacuac uatctttcac catcatcata tec 33

Claims (38)

1. CLAIMS 1. A Brassica plant that produces seeds having a long chain polyunsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% based on the total fatty acid composition.
2. 2. The plan of Claim 1, said seed comprising an oleic acid conlinide of at least about 37% based on the total fatty acid composition.
3. 3. The plant of Claim 1, said seed having an eicosoic acid content of at least about 14% based on the lofal fatty acid composition.
4. 4. The plan of Claim 1, wherein said monounsaturated fatty acid content is from about 85% to 90%.
5. 5. The plant of Claim 4, said seed having an oleic acid content of at least about 42% based on the total fatty acid composition.
6. 6. The plant of Claim 5, wherein said oleic acid content is from about 47% to about 56%.
7. 7. The plant of Claim 4, said seeds having an erucic acid content of from about 17% to about 31% based on the total fatty acid composition.
8. 8. The plant of Claim 4, said seeds having an eicosoic acid content of about 15% to about 21% based on the total fatty acid composition.
9. 9. The plant of Claim 1, said seeds having a saturated fatty acid conent less than 7% based on the total fatty acid composition.
10. 10. The plant of Claim 9, said seeds having a salted fatty acid content of less than 4% based on the total fatty acid composition.
11. The plant of Claim 10, said seeds having a total saturated fatty acid content of from about 2% to about 4% based on the total composition of the fatty acid.
12. 12. The plant of Claim 1, said seeds having a polyunsaturated fatty acid content of less than 1 1% based on the loyal fatty acid composition.
13. 13. The plant of Claim 12, said seeds having a polyunsaturated fatty acid content of from about 6% to about 11% based on the total fatty acid composition.
14. 14. The plan of Claim 1, said seeds having an α-linolenic acid content of from about 1% to about 2% based on the total fatty acid composition.
15. 15. The descendants of the plant of Claim 1, said offspring having said long chain polyunsaturated fatty acid content and said erucic acid content.
16. 16. A Brassica seed oil having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% based on the tofal fatty acid composition.
17. 17. The oil of Claim 16, said oil having an oleic acid content of at least about 37% based on the total fatty acid composition.
18. 18. The oil of Claim 16, said aceylene having an eicosoic acid conlinide of at least about 14% based on the total fatty acid composition.
19. 19. The oil of Claim 16, wherein said monounsaturated fatty acid conen is from about 85% to about 90%.
20. 20. The oil of Claim 19, said oil having an oleic acid content of at least about 42% based on the tolal fatty acid composition. twenty-one .
21. The oil of Claim 20, wherein said oleic acid content is about 47% has been about 56%.
22. 22. The oil of Claim 19, said oil having an erucic acid content of about 17% up to about 31% based on the total fatty acid composition.
23. 23. The oil of Claim 1 9, said oil having an eicosoic acid content of about 1 5% to about 21% based on the total composition of the fatty acid.
24. 24. The oil of Claim 16, said oil having a saturated fatty acid content of less than about 7% based on the total fatty acid composition.
25. 25. The aceil of Claim 16, said oil being a polyunsaturated fatty acid conlinide of less than 1 1% based on the total composition of the fatty acid.
26. 26. The oil of Claim 25, wherein said content of polyunsaturated fatty acid is less than 9%.
27. 27. A Brassica seed oil having a long chain monounsaturated fatty acid content of at least about 82%, wherein the sum of the nervonic acid, erucic acid, and eicosenoic acid condenide is approximately 50% up to approximately 66% based on the total fatty acid composition.
28. 28. The seed oil of Claim 27, wherein the oleic acid content is from about 25% to about 30%. 30.
29. A method for the production of a Brassica plant, said method comprising the steps of crossing a first plant line with a second plant line, and selecting the descendants of said crossing, wherein said first line of water yields an acid content. erucic of at least about 45% based on the total fatty acid composition and said second line of plants has an oleic acid content of at least about 84% based on the total composition of the fatty acid, said offspring having an long chain monounsaturated fatty acid content of at least about 82% and an erucic acid confetide of at least about 15% based on the total fatty acid composition.
30. 30. A method for making a vegetable oil, said method comprising the steps of grinding the Brassica seed having a long chain monounsaturated fatty acid conent of at least about 82% and an erucic acid content of at least about 15% based on the total composition of the fatty acid, and the extraction of said vegetable oil from said ground seeds.
31. 31 The method of Claim 30, which further comprises the steps of refining and bleaching said oil.
32. 32. The method of Claim 31, which further comprises the step of deodorizing said oil.
33. 33. A lubricant, comprising a Brassica oil having a long chain monounsaturated fatty acid content of at least about 82% and an erucic acid content of at least about 15% based on the total composition of the fatty acid , and an additive.
34. 34. The lubricant of Claim 33, wherein said additive is selected from the group consisting of an antioxidant, a rust inhibitor, a corrosion inhibitor, a pour point depressant, a foam additive, a colorant and a detergent.
35. 35. The lubricant of Claim 33, wherein said additive is present in an amount of about 0.01% to about 20% by weight.
36. 36. A hydraulic fluid comprising a Brassica oil having a long chain monounsaturated fatty acid conenide of at least about 82% and an erucic acid content of at least about 15% based on the total fatty acid composition , and an additive.
37. 37. The hydraulic fluid of Claim 36, wherein said additive is selected from the group consisting of an antioxidant, a rust inhibitor, a corrosion inhibitor, a pour point depressant, an amphoteric additive, a colorant and a detergent.
38. 38. The hydraulic fluid of Claim 36, wherein said additive is present in an amount of about 0.01% to about 20% by weight.