Method for assisting in identifying resistance of soybeans to soybean mosaic viruses

A technique for soybean mosaic virus and auxiliary identification, which is applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., can solve the problems of marker-gene genetic distance and small application range, and achieve saving Experimental cost and effect of saving experimental time

Inactive Publication Date: 2014-12-10
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Theoretically, only molecular markers with a genetic distance of less than 5 cM from disease-resistant genes have the application value of molecular-assisted selection. The marker-gene genetic distance obtained from previous research results is relatively long, and the scope of application is small

Method used

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  • Method for assisting in identifying resistance of soybeans to soybean mosaic viruses
  • Method for assisting in identifying resistance of soybeans to soybean mosaic viruses
  • Method for assisting in identifying resistance of soybeans to soybean mosaic viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1, the design and preparation of primer pair

[0056] Design and synthesize the following three pairs of primers:

[0057] SSR23 primer pair (F stands for upstream primer, R stands for downstream primer):

[0058] SSR23-F (sequence 1 of the sequence listing): 5'-TCGGTCCTATCCCCCTCCTAA-3';

[0059] SSR23-R (SEQ ID NO: 2 of the Sequence Listing): 5'-CGAGTCGCTTTTTCTCTTCTGT-3'.

[0060] SNP9 primer pair:

[0061] SNP9-F (sequence 3 of the sequence listing): 5'-GCTGGCTACACCAACGAAC-3';

[0062] SNP9-R (SEQ ID NO: 4 of the Sequence Listing): 5'-TATTAGAATGTGGACGGCAAA-3'.

[0063] SNP11 primer pair:

[0064] SNP11-F (sequence 5 of the sequence listing): 5'-TATCATGGCTAGTAGGCATAGATAGA-3';

[0065] SNP11-R (SEQ ID NO: 6 of the Sequence Listing): 5'-CCTTTATGAAAATTTTGTGATTGGAAC-3'.

Embodiment 2

[0066] Embodiment 2, the acquisition of the gene locus (RsmvN1-1) of the anti-SMV-N1 strain and the gene locus (RsmvN3-1) of the anti-SMV-N3 strain

[0067] 1. Construction of recombinant inbred lines

[0068] 1. The first generation of hybrids (F 1 generation).

[0069] 2. Put F 1 Generation self-pollination obtains 130 seeds, namely the second generation of children (F 2 generation).

[0070] 3. Put F 2 Each generation was subcultured 6 times by offspring single seed pass method (SSD), and 130 F 2：6 generation of recombinant inbred lines.

[0071] will be 130 F 2：6 Recombinant inbred lines, Dongnong 93-046 (female parent) and Hefeng 25 (male parent) were identified in Step 2 and Step 3, respectively.

[0072] 2. Identification of Plant Anti-SMV-N1 Strains and Anti-SMV-N3 Strains

[0073] The identification of the anti-SMV-N1 strain and the anti-SMV-N3 strain of the plants in each group were carried out in an independent aphid-proof net room. The experiments were ca...

Embodiment 3

[0089] Application of three pairs of primers prepared in embodiment 3 and embodiment 1 in the selection of hybrid offspring

[0090] 1. Construction of recombinant inbred lines

[0091] 1. Crossbreed Dongnong 93-046(♀) and conrad(♂) to obtain the first generation of hybrids (F 1 generation).

[0092] 2. Put F 1 Generation self-pollination obtains 150 seeds, namely the second generation of children (F 2 generation).

[0093] 3. Put F 2 Generations were subcultured for 6 times by single-seed seed transfer method (SSD) respectively, and 150 F 2：6 generation of recombinant inbred lines.

[0094] will be 150 F 2：6 Recombinant inbred lines, Dongnong 93-046 (female parent), conrad (male parent) and Hefeng 25 (susceptible control) were identified in Step 2 and Step 3, respectively.

[0095] 2. Identification of Plant Anti-SMV-N1 Strains and Anti-SMV-N3 Strains

[0096] The identification of the anti-SMV-N1 strain and the anti-SMV-N3 strain of the plants in each group were car...

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Abstract

The invention discloses a method for assisting in identifying the resistance of soybeans to soybean mosaic viruses. The invention protects a primer pair A consisting of a DNA fragment shown by a sequence 1 in the sequence table and a DNA fragment shown by a sequence 2 in the sequence table; the primer pair A have functions of (a) or (b) as follow: (a) assisting in identifying the resistance of soybeans to soybean mosaic viruses, and (b) assisting in selecting soybeans capable of resisting to soybean mosaic viruses. By adopting the method of the invention, molecular-assisted selective breeding of soybeans capable of resisting to soybean mosaic virus SMV-N1 strains and soybean mosaic virus SMV-N3 strains is realized, early-stages (such as a cotyledon stage) selective breeding with efficiency over 98% can be realized comparing with conventional disease-resistant phenotype identification method, and experiment time and experiment cost are saved greatly. The method of the invention will play an important role in selective breeding of soybeans.

Description

technical field [0001] The invention relates to a method for assisting identification of soybean resistance to soybean mosaic virus. Background technique [0002] Soybean (Glycine max) originates in China and is an important economic crop. The disease caused by soybean mosaic virus (SMV) is one of the main diseases that endanger soybean production, seriously affecting soybean yield and quality. Breeding disease-resistant varieties is currently the most effective way to control the prevalence of SMV. N1 strain and N3 strain were the dominant strains of SMV in soybean producing areas in Northeast China. Using molecular marker technology to locate genes related to N1 strain resistance and N3 strain resistance and closely linked molecular markers is beneficial to the breeding practice of molecular-assisted selection of resistant soybean varieties. [0003] Liu Lijun et al. (2002) used the F2 population of Heinong 39×Hefeng 25 to screen out the co-dominant RAPD marker OPN1400 / ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 赵雪滕卫丽韩英鹏李文滨李永光李冬梅朱聃
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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