Application of single nucleotide polymorphism site in auxiliary identification of resistance of soybean mosaic virus

A single nucleotide polymorphism, soybean mosaic virus technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problem that gene functions are not fully elucidated

Inactive Publication Date: 2017-10-03
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the soybean genome has been sequenced as early as 2010, the functions of each gene have not been fully elucidated

Method used

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  • Application of single nucleotide polymorphism site in auxiliary identification of resistance of soybean mosaic virus
  • Application of single nucleotide polymorphism site in auxiliary identification of resistance of soybean mosaic virus
  • Application of single nucleotide polymorphism site in auxiliary identification of resistance of soybean mosaic virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036]Example 1 Excavation of SNP sites in soybean gene Glyma.07G250300

[0037] Glyma.07G250300 in the Soybean Genome Database (url ) in the corresponding gene is GmZFP3 (See Chinese patent ZL201110233787.2), without introns, resequenced by Zhou et al. 302 soybean materials were found to contain a SNP site in the open reading frame (ORF) of Glyma.07G250300, in which there are two types of G / A at the 557th nucleotide position, and it is a sense mutation, and the amino acid is represented by R (arginine) to H(histidine) (as in figure 1 ).

Embodiment 2

[0038] Example 2 Analysis of the Glyma.07G250300 Gene Coding Sequence in Soybean Materials

[0039] The tested soybean materials are: Willimas82 (corresponding sequence is GmZFP3); wild soybean BB52 (GsZFP3), SA001, SA002, SA025, SA062, SA101, SA162, SA189, SA242 and Willimas82.

[0040] The above 10 kinds of soybeans ( Glycine max (L.) Merr.) The genomic DNA in the leaves of the seedlings of the species; using the DNA as a template, PCR amplification was performed with a primer pair, and the primer sequence (used to amplify the open reading frame of Glyma.07G250300) was:

[0041] The forward primer (F) is shown in SEQ ID NO.1, and the reverse primer is shown in SEQ ID NO.2;

[0042] PCR reaction system: 2×Phanta Max Buffer 25 μL; dNTP Mix (10mM each) 1 μL; upstream and downstream primers (F / R) 4 μL (10 μM); Phanta Max Super-Fidelity DNA Polymerase (1 U / μL); template DNA 4 μL ( 20ng / μL); ddH 2 O to make up to 50 μL.

[0043] The PCR amplification program was pre-denaturat...

Embodiment 3

[0046] Example 3 Mosaic Virus Resistance Identification of Soybean Materials

[0047] The soybean materials listed in Table 1 were inoculated and identified by using two virus strains SC3 and SC7 (provided by the Soybean Research Institute of Nanjing Agricultural University) that are prevalent in the south. The identification methods and evaluation indicators (resistance and sensitivity) refer to Nanjing Agricultural University in 2000 Wang Xiuqiang's master's degree thesis "Research on Soybean Mosaic Virus (SMV) Strain Identification, Resistance Source Screening and Resistance Genetics in Huanghuai and the Middle and Lower Reaches of the Yangtze River".

[0048] Table 1 Identification materials and results of soybean mosaic virus

[0049]

[0050] The results showed that BB52 and SA101 were resistant to both viruses, while SA062 was only resistant to SC3 and sensitive to SC7; while the rest of the materials were sensitive to both viruses (Table 1). Combined with the resul...

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Abstract

The invention discloses the application of the single nucleotide polymorphism of the 557th nucleotide in the open reading frame of the Glyma.07G250300 gene coding region in the soybean genome to assist in the identification of soybean mosaic virus resistance detection reagents; the site When the genotype is "G", the soybean material shows sensitivity, when the locus genotype is "A", the soybean material shows resistance; in practical applications, this locus can be used to design soybean mosaic virus resistance detection reagents , to evaluate the mosaic virus resistance of soybean materials at the soybean seedling stage to improve the efficiency of disease-resistant breeding, which has broad application prospects in actual breeding and production.

Description

technical field [0001] The invention relates to the technical fields of soybean disease-resistant breeding and molecular biology, in particular to an application of a SNP site related to soybean mosaic virus disease and a detection method for soybean mosaic virus resistance. Background technique [0002] Soybean Mosaic Virus (SMV) is a worldwide soybean disease and one of the viruses that have the greatest impact on soybean production, seriously affecting soybean yield and quality. Nanjing Agricultural University divided my country's SMV strains into SC1-SC21, among which SC3 and SC7 are the most popular strains in the soybean producing areas of the Huanghuai and Yangtze River basins. Soybean mosaic virus is widely distributed and harmful, and it is difficult to control it with chemical agents. Breeding and screening disease-resistant varieties is the most effective and economical way to solve soybean virus. [0003] Single nucleotide polymorphism (single nucleotide polymor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 张大勇宁丽华邵宏波徐照龙黄益洪刘佳戚维聪
Owner JIANGSU ACAD OF AGRI SCI
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