Mosaic-virus resistance GsCAD1 gene separated from wild soybeans, encoded protein and application of GsCAD1 gene

A wild soybean and mosaic virus technology, applied in the field of genetic engineering, can solve the problems of resistance loss of disease-resistant plants, etc., to achieve an improved level, ensure yield, and achieve long-term effective control effects

Active Publication Date: 2019-11-22
NORTHEAST NORMAL UNIVERSITY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

People prevent and control soybean mosaic virus disease mainly by planting disease-resistant soybean varieties. However, due to the diversity of soybean mosaic virus physiological r

Method used

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  • Mosaic-virus resistance GsCAD1 gene separated from wild soybeans, encoded protein and application of GsCAD1 gene
  • Mosaic-virus resistance GsCAD1 gene separated from wild soybeans, encoded protein and application of GsCAD1 gene
  • Mosaic-virus resistance GsCAD1 gene separated from wild soybeans, encoded protein and application of GsCAD1 gene

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Experimental program
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Effect test

Embodiment

[0026] 1. Construction of recombinant plasmids

[0027] 1. Extract the RNA of wild soybean BYO-5, and use the reverse transcription kit (AT311-03) to obtain the cDNA of wild soybean BYO-5.

[0028] 2. Using primer F1: CTCTAGA ATGGCAGCACAAGCTGAA (underlined is the restriction site XbaI), primer R1: CGGATCC AATTTCAGTGTGTTTCCA (underlined is the restriction site BamHI), using the cDNA of wild soybean BYO-5 as a template for PCR, the reaction system is shown in Table 1, and the PCR amplification procedure is shown in Table 2

[0029] Table 1 Amplification system

[0030]

[0031] Table 2 PCR amplification program

[0032]

[0033]

[0034] The 50uL PCR product was detected on 1.0% agarose gel electrophoresis and a PCR fragment with a size of about 1080 was recovered.

[0035] 3. Clone the wild soybean GSCAD1 gene and connect it to the pMD-18T vector (the reaction system is shown in Table 3 below)

[0036] Table 3 Reaction system for linking pMD18-T vector

[0037]...

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Abstract

The invention discloses a mosaic-virus resistance GsCAD1 gene separated from wild soybeans. The nucleotide sequence of the GsCAD1 gene is shown in SEQ No.1 and closely related to mosaic virus resistance. The invention further discloses a protein encoded by the GsCAD1 gene. The amino acid sequence of the protein is shown in SEQ No.2. The invention also discloses application of the GsCAD1 gene. Whenin use, the GsCAD1 gene is introduced into a target plant to obtain a transgenic plant of which the plant mosaic-virus resistance is higher than that of the target plant. Overexpression of the GsCAD1gene can significantly improve the mosaic-virus disease resistance of the soybeans, a method for permanently and effectively preventing and treating soybean mosaic-virus diseases is provided, and thus the yield of the soybeans and economic benefits of farmer households are ensured.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to an anti-mosaic virus nucleotide sequence isolated from wild soybeans. Background technique [0002] Soybean Mosaic Virus (SMV) disease is one of the important diseases that seriously affect the yield and seed quality of soybeans in my country and often occurs in major soybean producing areas in China. It seriously affects the yield of soybeans in my country. Under natural conditions, it can cause 8-50% yield loss of soybean in our country. The in vitro preservation period of SMV is about 4-5 days, 15 days and 120 days at room temperature, 4°C and below 0°C, respectively. The lethal temperature of SMV is 58-66℃, and the dilution limit is 10-5. SMV can be transmitted between generations and different regions through seeds, and can be non-persistently transmitted in the field through sap friction inoculation and aphids. [0003] SMV is a single-stranded positive-sense ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/82A01H5/00A01H6/54
CPCC07K14/415C12N15/8283Y02A40/146
Inventor 姜丽丽庞劲松寻红卫王蒙王莹于佳淼杨向东郭东全杨静牛路钱雪艳邢国杰董英山李启云刘宝
Owner NORTHEAST NORMAL UNIVERSITY
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