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Fatty acid desaturases and mutant sequences thereof

a technology of fatty acid desaturase and mutant sequences, which is applied in the field of fatty acid desaturases and nucleic acids encoding desaturase proteins, can solve the problems of low -linolenic acid mutation derived from the rakow mutation breeding program that did not have direct commercial application, and no teachings that disclose mutations in delta-12 or delta-15 fatty acid desaturase coding sequences from plants

Inactive Publication Date: 2007-07-12
DEBONTE LORIN R +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The invention is about controlling the levels of unsaturated fatty acids in plants through mutations in genes. The patent describes isolated nucleic acid fragments that contain mutations in either the delta-12 or delta-15 fatty acid desaturase gene. The mutations can be introduced into a variety of Brassicaceae or Helianthus species, such as soybean, oilseed Brassica species, sunflower, castor bean or corn. The resulting plant lines can have altered fatty acid compositions, with higher levels of unsaturated fatty acids. The invention also includes a method of producing plant lines with both a delta-12 and a delta-15 desaturase mutation. The resulting vegetable oil has an altered fatty acid composition."

Problems solved by technology

The low α-linolenic acid mutation derived from the Rakow mutation breeding program did not have direct commercial application because of low seed yield.
However, there are no teachings that disclose mutations in delta-12 or delta-15 fatty acid desaturase coding sequences from plants.

Method used

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  • Fatty acid desaturases and mutant sequences thereof
  • Fatty acid desaturases and mutant sequences thereof
  • Fatty acid desaturases and mutant sequences thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Mutagenesis

[0073] Seeds of Westar, a Canadian (Brassica napus) spring canola variety, were subjected to chemical mutagenesis. Westar is a registered Canadian spring variety with canola quality. The fatty acid composition of field-grown Westar, 3.9% C16:0, 1.9% C18:1, 67.5% C18:1, 17.6% C18:2, 7.4% C18:3, 22:1, has remained stable under commercial production, with <±10% deviation, since 1982.

[0074] Prior to mutagenesis, 30,000 seeds of B. napus cv. Westar seeds were preimbibed in 300-seed lots for two hours on wet filter paper to soften the seed coat. The preimbibed seeds were placed in 80 mM ethylmethanesulfonate (EMS) for four hours. Following mutagenesis, the seeds were rinsed three times in distilled water. The seeds were sown in 48-well flats containing Pro-Mix. Sixty-eight percent of the mutagenized seed germinated. The plants were maintained at 25° C. / 15° C., 14 / 10 hr day / night conditions in the greenhouse. At flowering, each plant was individually self-pollinated.

[0075] M2...

example 2

High Oleic Acid Canola Lines

[0087] In the studies of Example 1, at the M3 generation, 31 lines exceeded the upper statistical threshold for oleic acid (≧71.0%). Line W7608.3 had 71.2% oleic acid. At the M4 generation, its selfed progeny (W7608.3.5, since designated A129.5) continued to exceed the upper statistical threshold for C18:1 with 78.8% oleic acid. M5 seed of five self-pollinated plants of line A129.5 (ATCC 40811) averaged 75.0% oleic acid. A single plant selection, A129.5.3 had 75.6% oleic acid. The fatty acid composition of this high oleic acid mutant, which was stable under both field and greenhouse conditions to the M, generation, is summarized in Table 9. This line also stably maintained its mutant fatty acid composition to the M7 generation in field trials in multiple locations. Over all locations the self-pollinated plants (A129) averaged 78.3% oleic acid. The fatty acid composition of the A129 for each Idaho trial location are summarized in Table 10. In multiple loc...

example 3

Low Linoleic Acid Canola

[0095] In the studies of Example 1, at the M3 generation, 80 lines exceeded the lower statistical threshold for linoleic acid (≦13.2%). Line W12638.8 had 9.4% linoleic acid. At the M4 and M5 generations, its selfed progenies [W12638.8, since designated A133.1 (ATCC 40812)] continued to exceed the statistical threshold for low C18:2 with linoleic acid levels of 10.2% and 8.4%, respectively. The fatty acid composition of this low linoleic acid mutant, which was stable to the M7 generation under both field and greenhouse conditions, is summarized in Table 13. In multiple location replicated yield trials, A133 was not significantly different in yield from the parent cultivar Westar. An additional low linoleic acid line, designated M3062.8 (ATCC 75025), was also produced by the disclosed method. A 10-seed bulk analysis of this line showed the following fatty acid composition: C16:0—3.8%, C18:0—2.3%, C18:1—77.1%, C18:2—8.9%, C18:3—4.3%, FDA Sats—6.1%. This line ha...

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Abstract

Seeds, plants and oils are provided having high oleic acid; low linoleic acid; and low linoleic acid plus linolenic acid; and advantageous functional or nutritional properties. Plants are disclosed that contain a mutation in a delta-12 or delta-15 fatty acid desaturase gene. Preferred plants are rapeseed and sunflower plants. Plants carrying such mutant genes have altered fatty acid composition in seeds. In one embodiment, a plant contains a mutation in a region having the conserved motif His-Xaa-Xaa-Xaa-His, found in delta-12 and delta-15 fatty acid desaturases. A preferred motif has the sequence His-Glu-Cys-Gly-His. A preferred mutation in this motif has the amino acid sequence His-Lys-Cys-Gly-His. Nucleic acid fragments are disclosed that comprise a mutant delta-12 or delta-15 fatty acid desaturase gene sequence.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is a continuation-in-part of PCT application Ser. No. PCT / US96 / 20090, filed Dec. 13, 1996, which is a continuation-in-part of U.S. application Ser. No. 08 / 572,027, filed Dec. 14, 1995.TECHNICAL FIELD [0002] This invention relates to fatty acid desaturases and nucleic acids encoding desaturase proteins. More particularly, the invention relates to nucleic acids encoding delta-12 and delta-15 fatty acid desaturase proteins that affect fatty acid composition in plants, polypeptides produced from such nucleic acids and plants expressing such nucleic acids. BACKGROUND OF THE INVENTION [0003] Many breeding studies have been conducted to improve the fatty acid profile of Brassica varieties. Pleines and Freidt, Fat Sci. Technol., 90(5), 167-171 (1988) describe plant lines with reduced C18:3 levels (2.5-5.8%) combined with high oleic content (73-79%). Rakow and McGregor, J. Amer. Oil Chem. Soc., 50, 400-403 (Oct. 1973) discuss pro...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01H5/00C07H21/04C12N9/04C12N15/82C07C53/126C07C57/02C07C57/03C12N9/02
CPCC07C53/126C07C57/02C12N15/8247C12N9/0083C07C57/03
Inventor DEBONTE, LORIN R.FAN, ZHEGONGMIAO, GUO-HUA
Owner DEBONTE LORIN R
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