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NAC transcription factor gene GmST2 of soybean holy bean No.9 and application of NAC transcription factor gene GmST2

A transcription factor and gene technology, applied in soybean Shengdou No. 9 NAC transcription factor gene - GmST2 and its application field, to achieve the effect of improving salt/drought tolerance and improving salt/drought tolerance

Active Publication Date: 2012-09-19
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the function of soybean NAC membrane-bound transcription factor

Method used

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  • NAC transcription factor gene GmST2 of soybean holy bean No.9 and application of NAC transcription factor gene GmST2
  • NAC transcription factor gene GmST2 of soybean holy bean No.9 and application of NAC transcription factor gene GmST2
  • NAC transcription factor gene GmST2 of soybean holy bean No.9 and application of NAC transcription factor gene GmST2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Cloning of embodiment 1, GmST2

[0029] 1.1 Extraction of total RNA of Shengdou No. 9

[0030] (1) Put the plant material of Shengdou No. 9 into a mortar, and use liquid nitrogen to grind it into powder (directly used in the following experiments or frozen in a -80°C ultra-low temperature refrigerator for later use);

[0031] (2) After the liquid nitrogen volatilizes, immediately transfer 100-200mg of plant powder into a 1.5ml centrifuge tube, then quickly add 1ml of Trizol extract, vortex to fully dissolve the sample into the extract, and place it at room temperature for 5 minutes;

[0032] (3) 4°C, 12,000rpm, centrifuge for 10min, transfer 0.9ml of the supernatant to a new 1.5ml centrifuge tube, then add 0.2ml of chloroform, vigorously shake and mix for 15sec, and place at room temperature for 2-5min;

[0033] (4) 4°C, 12,000rpm, centrifuge for 10min, transfer 0.4ml of supernatant to a new 1.5ml centrifuge tube, add 0.4ml of isopropanol, turn up and down 15 times to...

Embodiment 2

[0138] Example 2, Functional Verification of Expressing the NAC Transcription Factor Gene GmST2 of Saint Bean No. 9 in Arabidopsis

[0139] 2.1 Transformation of Arabidopsis thaliana by flower infection method

[0140] (1) When Arabidopsis thaliana (Col-0 wild type) grows to 1 cm of bolting, the top is subtracted to induce the formation of lateral inflorescences;

[0141] (2) One day before transformation, take 1ml of activated Agrobacterium GV3101 containing the expression vector plasmid and add it to 40ml of YEP medium containing corresponding antibiotics and 50μg / ml rifampicin, and culture with shaking at 28°C until OD 600 About 1.0-1.2;

[0142] (3) room temperature, 4200rpm, centrifuge 10min, collect the thalli, resuspend the thalli with the liquid (5% sucrose, 0.05% Silwet L-77), make OD 600 about 0.8;

[0143] (4) Use a pipette to drop the Agrobacterium on the inflorescences for dipping, and after all the inflorescences are infected, put the Arabidopsis thaliana into...

Embodiment 3

[0182] Example 3, Functional Verification of Expressing Shengdou No. 9 NAC Transcription Factor Gene GmST2 in Soybean

[0183] 3.1 Transformation of Soybean by Vacuum Infiltration Assisted Transformation of Germinated Soybean Embryos

[0184] (1) Seed disinfection and pre-cultivation treatment

[0185] Soybean seeds were sterilized with 70% ethanol for 5 minutes, and then cleaned with 0.1% HgCl 2 Disinfect for 10 minutes, rinse with sterile water 5-6 times, and soak the seeds in sterile water for 12 hours at 25°C-28°C.

[0186] Soybeans whose embryos swelled and germinated after soaking were put into the pre-cultivation medium, and cultured in light at 28°C for 1 day, wherein the formula of the pre-culture medium was: MS+3.0mg / L 6-BA (6-benzylaminoadenine) +20g / L sucrose+7g / L agar, pH5.8.

[0187] (2) After the seed embryo swells and germinates, it is cut to expose or damage the germinated embryo part

[0188] When the whole pre-cultivated soybean embryo is enlarged and th...

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Abstract

The invention discloses an NAC (N-acetyl-L-Cysteine) transcription factor gene GmST2 of soybean holy bean No.9 and a plant expression vector of the GmST2. The invention further discloses application of the gene GmST2 in improvement on the salt resistant / drought tolerance in arabidopsis thaliana and soybean plants. Provided by tests, the salt resistance / drought tolerance of a transgenic plant is greatly improved compared with that of a non transgenic plant and theoretical base and practice foundation for improving the salt resistance / drought tolerance of crops through genetic engineering means are provided. The NAC transcription factor gene GmST2 can be widely applied to cultivation of salt-resisting / drought tolerant plant varieties.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and in particular relates to soybean Shengdou No. 9 NAC transcription factor gene—GmST2 and its application. Background technique [0002] Soybean is an important economic crop, so improving and optimizing soybean quality has become a focus of attention. my country is one of the most water-deficient countries in the world. Drought is the main natural disaster in my country's agricultural production, and the area of ​​saline-alkali land and secondary salinized cultivated land is increasing year by year. These unfavorable environmental conditions seriously restrict my country's crop production. In order to adapt to these unfavorable environments, plants develop a series of internal defense measures, which also require the participation of abiotic stress response genes. Many salt- and drought-responsive genes have been identified and their functions in abiotic stress verified, but th...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/84A01H5/00
Inventor 向凤宁姬丹丹李朔
Owner SHANDONG UNIV
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