Method for increasing the total oil content in oil plants

A technology for oil content and oil crops, which can be applied to biochemical equipment and methods, the use of carriers to introduce foreign genetic material, oxidoreductase, etc., and can solve problems such as increased oil content.

Inactive Publication Date: 2009-01-28
MAX PLANCK GESELLSCHAFT ZUR FOERDERUNG DER WISSENSCHAFTEN EV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0021] None of the currently disclosed methods of increasing oil content in transgenic plants resulted in an increase in oil content in cultivable plants sufficient for technical manipulation

Method used

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  • Method for increasing the total oil content in oil plants
  • Method for increasing the total oil content in oil plants
  • Method for increasing the total oil content in oil plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0214] Example 1: Cloning of yeast Gpd1 gene

[0215] Genomic DNA was isolated from Saccharomyces cerevisiae S288C (Mat alpha SUC2 malmel gal2 CUP1 flo1 flo8-1; Invitrogen, Karlsruhe, Germany) following the protocol disclosed below:

[0216] A 100 ml culture was grown at 30°C to an optical density of 1.0. 60 ml of the culture was centrifuged at 3000 xg for 3 minutes. Resuspend the pellet in 6 ml double distilled HO 2 O, and the suspension was divided into 1.5ml containers and centrifuged, and the supernatant was discarded. The pellet was resuspended by vortexing in 200 μl solution A, 200 μl phenol / chloroform (1:1) and 0.3 g glass beads before lysing the cells. After adding 200 μl of TE buffer (pH 8.0), the lysate was centrifuged for 5 minutes. The supernatant was ethanol precipitated using 1 ml of ethanol. After precipitation, the obtained pellet was dissolved in 400 μl TE buffer pH 8.0 + 30 μl / ml RNaseA. After incubation at 37°C for 5 minutes, 18 µl of 3 M sodium acetat...

Embodiment 2

[0250] Example 2: Plasmids for Transformation of Plants

[0251] Binary vectors such as pBinAR can be used to transform plants ( and Willmitzer (1990) Plant Science 66:221-230). The binary vector can be constructed by ligating cDNA into T-DNA in sense or antisense orientation. A plant promoter located 5' to the cDNA activates transcription of the cDNA. The polyadenylation sequence is located 3' of the cDNA.

[0252] Tissue-specific expression can be obtained using tissue-specific promoters. Seed-specific expression can be obtained, for example, by cloning a rapeseed protein or a LeB4- or USP promoter into the 5' end of the cDNA. Any other seed-specific promoter can also be used. The CaMV 35S promoter can be used for constitutive expression throughout the plant.

[0253] Further examples of binary vectors are the vectors pSUN-USP and pGPTV-rapeseed protein, into which the fragment of Example 2 was cloned. Vector pSUN-USP contains USP promoter and OCS terminator. The ve...

Embodiment 3

[0255] Example 3: Transformation of Agrobacterium

[0256] Agrobacterium-mediated transformation of plants can be performed, for example, using Agrobacterium tumefaciens strain GV3101 (pMP90) (Koncz and Schell (1986) Mol Gen Genet 204:383-396) or LBA4404 (Clontech). Transformation can be performed using standard transformation techniques (Deblaere et al. (1984) Nucl Acids Res 13:4777-4788).

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Abstract

The invention relates to methods for increasing the total oil content and/or the content of glycerol-3-phosphate in transgenic oil plants, which contain at least 20 wt.-% oleic acid in relation to the total fatty acid content, preferably in plant seeds, by the expression of glycerol-3-phosphate dehydrogenases (G3PDH) from yeasts, preferably from Saccharomyces cerevisiae. Advantageously, the oil obtained by this method and/or the free fatty acids are added to polymers, foodstuffs and feedstuffs, cosmetics, pharmaceutical products or products with industrial applications.

Description

technical field [0001] The present invention relates to a method for increasing total oil content and / or glycerol 3-phosphate content in transgenic oil crops by expressing glycerol 3-phosphate dehydrogenase (G3PDH) from yeast, preferably Saccharomyces cerevisiae The crop comprises at least 20% by weight oleic acid of the total fatty acid content, preferably in plant seeds. The oil and / or free fatty acid obtained in this method can be conveniently added in polymers, food, feed, cosmetics, pharmaceuticals or products with industrial applications. Background technique [0002] Increasing the total oil content and / or the glycerol 3-phosphate content in transgenic oil crops, especially plant seeds, is of great importance for traditional and modern plant breeding, and especially for plant biotechnology. Due to the increasing consumption of vegetable oils for nutritional or industrial applications, the possibility of enhancing or modifying vegetable oils is increasingly the subjec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N9/04
CPCC12N15/8247C12N9/0006
Inventor T·灿克O·奥斯瓦德J·鲍尔H·维格奥拉斯P·盖根贝格尔P·瓦尔德克M·斯蒂特
Owner MAX PLANCK GESELLSCHAFT ZUR FOERDERUNG DER WISSENSCHAFTEN EV
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