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Application of soybean protein and coding gene of the soybean protein in plant drought resistance adjustment

A technology encoding gene and transgenic plant is applied in the application field of soybean GmSYP24 protein and its encoding gene in regulating plant drought resistance, and can solve the problems of no drought resistance, high salt transgenic soybean, etc.

Active Publication Date: 2014-10-15
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Since Hinchee and others obtained glyphosate-resistant soybeans in 1988, Monsanto Corporation of the United States has obtained herbicide-resistant soybeans with different genes, which have been industrialized on a large scale. in production

Method used

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  • Application of soybean protein and coding gene of the soybean protein in plant drought resistance adjustment
  • Application of soybean protein and coding gene of the soybean protein in plant drought resistance adjustment
  • Application of soybean protein and coding gene of the soybean protein in plant drought resistance adjustment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1, the cloning of soybean GmSYP24 gene

[0040] The total RNA of soybean (Glycine max (L.) Merr.) variety Jindou 21 was extracted by Trizol method, and reverse-transcribed into cDNA, which was used as a template for PCR amplification using the following primers.

[0041] Upstream primer: 5'-ACATCATTTCCCACTCCCG-3' (position 1-19 of Sequence 1);

[0042] Downstream primer: 5'-GGACCTATGTCCAAAAGTAACCA-3' (reverse complement of positions 753-775 of Sequence 1).

[0043] Reaction system: template cDNA 2μL, upstream primer (10μmol / L) 0.5μL, downstream primer (10μmol / L) 0.5μL, 10×Buffer 2μL, magnesium chloride (25mM) 1.2μL, dNTPs (2.5mM) 0.4μL, DNA polymerase (5U / μL) 0.2 μL, make up to 20 μL with sterile water.

[0044] Reaction program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 0.5 min, annealing at 58°C for 0.5 min, extension at 72°C for 1 min, 30 cycles; final extension at 72°C for 10 min; storage at 4°C.

[0045] After the reaction, the tar...

Embodiment 2

[0047] Embodiment 2, the acquisition and identification of GmSYP24 transgenic tobacco

[0048] 1. Obtaining GmSYP24 transgenic tobacco

[0049] 1. Construction of recombinant expression vector pCXSN-GmSYP24

[0050] The pCXSN vector was digested with restriction endonuclease Xcm I to obtain a linear TA cloning vector with a 3' protruding T base. The digested pCXSN linear vector was ligated with the purified and recovered PCR product (sequence 1) obtained in Example 1.

[0051] Ligation system (10 μL): 1 μL of digested pCXSN vector, 4 μL of purified and recovered PCR product, and 5 μL of DNA ligase. Wherein, DNA ligase is a product of Takara Company, and its product catalog number is 6024.

[0052] React overnight at 16°C.

[0053] Transform the competent cells of Escherichia coli DH5α with 10 μL of the ligation product, spread evenly on the LB plates containing kanamycin, and incubate them upside down at 37°C for 12-16 hours. Pick the monoclonal colony on the transformati...

Embodiment 3

[0075] Embodiment 3, the drought resistance identification of GmSYP24 transgenic tobacco

[0076] The drought resistance of the five GmSYP24 transgenic tobacco homozygous plants TL1, TL2, TL9, TL10 and TL11 in Example 2 was identified according to the following method. The specific operation is as follows:

[0077] 1. Sterilize the seeds of 5 GmSYP24 transgenic tobacco homozygous plants TL1, TL2, TL9, TL10 and TL11, the pCXSN empty vector (TL0), and the non-transgenic wild-type tobacco (Nicotiana benthaminana.) In 1 / 2MS medium, culture in long-day light (16h light / 8h dark) at 22°C for 12 days. Each tobacco plant was supplied with normal water under the same conditions.

[0078] 2. Transplant each tobacco seedling into a nutrient soil for long-day sunshine (16h light / 8h dark), and cultivate under light at 22°C for 15 days. Each tobacco plant was supplied with normal water under the same conditions.

[0079] 3. Observe the phenotype of each tobacco plant, and take pictures a...

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Abstract

The invention discloses application of soybean GmSYP24 protein and a coding gene of the soybean protein in plant drought resistance adjustment, and particularly application of a protein as shown in sequence 2 and a coding gene of the protein in plant drought resistance adjustment. Experiments show that under normal water supply conditions, wild type tobacco, empty vector transduced tobacco and GmSYP24 transgenic tobacco lines are in good growth conditions, but tobacco leaves of the GmSYP24 transgenic tobacco lines are greener than that of the wild type tobacco and the empty vector transduced tobacco; under drought stress conditions, the tobacco leaves of the wild type tobacco and the empty vector transduced tobacco are easy to wither and yellow than that of the transgenic tobacco lines; the transgenic tobacco plants are better in growth conditions than the wild type tobacco and the empty vector transduced tobacco. The application suggests that soybean GmSYP24 gene is a potential soybean drought resistant gene, and lays a foundation for subsequent overexpression of the gene in soybean to obtain a drought resistant transgenic soybean new variety. The application of the soybean GmSYP24 protein and the coding gene has vital significance to alleviate the food crisis, save water resources, reduce the agricultural irrigation cost and the like.

Description

technical field [0001] The invention belongs to the field of genetic engineering and relates to the application of soybean GmSYP24 protein and its encoding gene in regulating plant drought resistance. Background technique [0002] Drought is a long-standing worldwide problem. The global arid and semi-arid areas account for about 35% of the land area, covering more than 60 countries and regions in the world. China has nearly 2.5×10 6 hm 2 Arable land is affected by drought. In particular, North China, Northeast China, and Northwest China, the main grain production areas in China, happen to be in areas with severe water shortages in China, resulting in a substantial reduction in crop production. Drought has become one of the main factors inducing the food crisis. According to the IPCC AR4 multi-model forecast of drought changes in China: From 2011 to 2050, China showed a continuous trend of drought, and the overall drought area and drought frequency continued to increase....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00
CPCC07K14/415C12N15/8273
Inventor 陈李淼沙爱华张婵娟单志慧陈水莲陈海峰邱徳珍周蓉周新安
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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