Application of anti-soybean mosaic virus gene GmRDBM

A soybean mosaic virus, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, to ensure the yield and quality, improve resistance, improve the effect of supply rate

Pending Publication Date: 2019-04-26
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Few genes have been shown to be

Method used

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  • Application of anti-soybean mosaic virus gene GmRDBM
  • Application of anti-soybean mosaic virus gene GmRDBM
  • Application of anti-soybean mosaic virus gene GmRDBM

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: GmR DBM Gene acquisition

[0019] 1. Extraction of RNA:

[0020] The soybean variety Hemp hemp was sown, and young leaves were collected after emergence for the extraction of total RNA. Put 0.1g of the above-mentioned leaves into a centrifuge tube, add liquid nitrogen to grind quickly, then add 1ml Trizol (Invitrogen Co., Ltd.) to mix and let stand for 5min; add 0.2ml of chloroform to shake for 15sec, let stand for 2min; centrifuge at 12000rpm 10min, absorb 400μl supernatant and add it to 400μl isopropanol and mix well; centrifuge for 10min, discard the supernatant, add 1ml of 70% ethanol and vortex for 1min; centrifuge for 5min, discard the supernatant, and put it in a fume hood to dry; add 50 μl ddH 2 O spare.

[0021] 2. Synthesis of first-strand cDNA:

[0022] Synthesis of first-strand cDNA was performed using PrimeScript II RTase (TAKARA). Pipette 1 μl of the above RNA into a PCR tube, add 1 μl each of Oligo dT Primer and dNTP Mixture, and add ddH ...

Embodiment 2

[0025] Example 2: GmR DBM Proof of gene resistance function

[0026] 1. Soybean Variety Hemp GmR DBM Obtaining knockout plants

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Abstract

The invention discloses application of anti-soybean mosaic virus gene GmRDBM. A gene GmRDBM for encoding a CC-NBS-LRR functional domain is cloned from big white gastrodia of an anti-soybean mosaic virus variety. The gene is sequenced, and the amino acid sequence encoded by the gene is obtained by means of software prediction. A Crispr/Cas9 technology is used for editing the GmRDBM sequence of thebig white gastrodia, and plants knocked out of GmRDBM are inoculated with a soybean mosaic virus for carrying out an experiment; the gene is proved to be the anti-soybean mosaic virus gene of the bigwhite gastrodia variety. The obtained gene and a protein sequence thereof can be used for the improvement of resistance of soybeans to the soybean mosaic virus.

Description

technical field [0001] The invention belongs to the technical field of biological genes, and relates to an anti-soybean mosaic virus gene GmR DBM Applications. technical background [0002] Soybean is the main source of edible oil and protein in my country and even in the world. Its production is affected by various diseases. Among them, soybean mosaic virus disease is one of the main diseases of soybean. Soybean mosaic virus has been found in all soybean producing areas of the country, and is divided into multiple strains according to the difference in infectivity; the soybean yield loss caused by soybean mosaic virus can reach 15% to 70%; at the same time, soybean mosaic virus It can also cause mottled seed coats of soybean seeds, reducing the quality. Since there is no effective agent to kill the virus at present, cultivating disease-resistant varieties is the most economical and effective way to control the disease, and obtaining disease-resistant genes is the materi...

Claims

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Application Information

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IPC IPC(8): C12N15/29A01H5/00A01H6/54
CPCC12N15/8283C07K14/415
Inventor 智海剑殷金龙王丽群晋彤彤李凯
Owner NANJING AGRICULTURAL UNIVERSITY
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