Single nucleotide polymorphism (SNP) molecular marker for resistance identification of cucumber green mottle mosaic virus disease in watermelon and application of molecular marker

A green mottled flower and molecular marker technology, applied in the direction of recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems affecting the high-throughput screening and identification of watermelon resistance to cucumber green mottle mosaic virus disease, etc. To achieve the effect of improving resistance and overcoming the long cycle

Active Publication Date: 2020-11-20
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the lack of research on germplasm identification, resistance genetics, and resistance gene-linked molecular markers against cucumber green mottle m

Method used

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  • Single nucleotide polymorphism (SNP) molecular marker for resistance identification of cucumber green mottle mosaic virus disease in watermelon and application of molecular marker
  • Single nucleotide polymorphism (SNP) molecular marker for resistance identification of cucumber green mottle mosaic virus disease in watermelon and application of molecular marker
  • Single nucleotide polymorphism (SNP) molecular marker for resistance identification of cucumber green mottle mosaic virus disease in watermelon and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1. Resistance identification of watermelon wild germplasm 'P.I.595203' and cultivar 'M1511-3' to cucumber green mottle mosaic virus disease:

[0076] Select watermelon materials --- 'P.I.595203', 'M1511-3', 'P.I.595203' and 'M1511-3' hybrid F1 offspring from the laboratory germplasm resource bank, inoculate with cucumber green mottle virus, and use the plant phenotype symptoms Determine resistance; if figure 1 .

[0077] details as follows:

[0078] 1. Cucumber green mottle mosaic virus inoculation:

[0079]Carry out inoculation at the stage of one leaf and one heart leaf of watermelon seedling, take by weighing the fresh diseased leaf 2g that has been infected with cucumber green mottle mosaic virus, put into the mortar of high temperature sterilization, according to 1:10 (W / V, namely, 1g: 10ml) was added into a corresponding volume of 0.02mol / LPBS buffer solution (pH7.2) and ground into a slurry, which was used as virus juice for artificial friction inocula...

Embodiment 2

[0090] Embodiment 2, genetic analysis and gene mapping of resistance to cucumber green mottled mosaic virus disease:

[0091] Selected melon materials from the germplasm resource bank of the laboratory --- 'P.I.595203', 'M1511-3', 'P.I.595203' and 'M1511-3' hybrid F1 offspring and F2 population obtained by selfing of F1, cucumber green mottled Mosaic virus inoculation, use plant phenotype symptoms to determine resistance, and carry out genetic analysis of disease resistance. Then based on the BSA method for disease resistance gene mapping, such as figure 2 .

[0092] Genetic analysis of resistance: the cucumber green mottled mosaic virus inoculation method in Example 1 was used to inoculate 304 individual plants of the F2 population, and the resistance was determined according to the phenotype, and the F2 population was resistant to the cucumber green mottled mosaic virus disease Sexual segregation occurred, including 33 disease-resistant plants and 271 susceptible plants, ...

Embodiment 3

[0102] Embodiment 3, the development and verification of marking CGMMVR with KASP

[0103] Watermelon materials --- 'P.I.595203', 'M1511-3', 'P.I.595203' and 'M1511-3' hybrid F1 offspring, F2 population obtained by F1 self-crossing and other 184 accessions were selected from the laboratory germplasm resource bank For watermelon germplasm, the SNPs were first transformed into KASP markers, and the genotype and genetic segregation rules were identified by amplifying the KASP marker CGMMVR with primers.

[0104] details as follows:

[0105] 1. KASP marks the development of CGMMVR:

[0106] 1. SNP information extraction:

[0107] According to the gene mapping results in Example 2, the SNP information closely linked with the disease resistance gene was extracted.

[0108] 2. Extract DNA

[0109] The genomic DNA of the hybrid F1 offspring of 'P.I.595203', 'M1511-3', 'P.I.595203' and 'M1511-3' was extracted, and the method was the same as in Example 2.

[0110] 3. PCR amplificat...

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Abstract

The invention discloses a molecular marker developed based on single nucleotide polymorphism (SNP) linked to resistance to a cucumber green mottle mosaic virus disease, and the molecular marker is used for resistance identification of the cucumber green mottle mosaic virus disease; a watermelon is used as a species, and molecular marker primers CGMMVR include an allelic primer 1, an allelic primer2 and a universal primer. The molecular marker can be used for identification of resistance of the germplasm to the cucumber green mottle mosaic virus disease in the watermelon and assistant selective breeding of hybrid or backcross descendants of the germplasm.

Description

technical field [0001] The invention belongs to the technical field of vegetable disease-resistant molecular marker development and molecular marker-assisted breeding, and specifically relates to the development and application of a molecular marker for watermelon resistance to cucumber green mottle mosaic virus. Background technique [0002] Cucumber green mottle mosaic virus (CGMMV) is a positive single-stranded RNA virus with a straight rod shape and belongs to the genus Tobamovirus. It was first reported in the UK in 1935 ( Ainsworth, 1935). It mainly infects cucurbit crops, such as watermelon, melon, cucumber, pumpkin, gourd, etc., and also infects mandala, amaranth, tobacco, and common tobacco. Different strains of CGMMV are on different host plants Symptoms are different. The watermelon strain CGMMV can cause watermelon mosaic, mottled, protruding leaves, dwarfing of plants and fruit collapse. CGMMV has strong adaptability and stress resistance. It can survive for s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2600/172C12Q2531/113C12Q2563/107
Inventor 杨景华张明方蔡玲敏胡仲远
Owner ZHEJIANG UNIV
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