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Two molecular markers for identifying vine blight resistance of muskmelons on basis of anti-vine blight linkage gene development and application of two molecular markers

A molecular marker and vine blight technology, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. The problem of long genetic distance, etc., to achieve the effect of improving the resistance and stability of vine blight

Active Publication Date: 2019-05-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the long genetic distance between the molecular marker and the disease resistance gene, and the traditional random DNA molecular marker, it affects the high-throughput screening and identification of muskmelon resistance to vine blight

Method used

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  • Two molecular markers for identifying vine blight resistance of muskmelons on basis of anti-vine blight linkage gene development and application of two molecular markers
  • Two molecular markers for identifying vine blight resistance of muskmelons on basis of anti-vine blight linkage gene development and application of two molecular markers
  • Two molecular markers for identifying vine blight resistance of muskmelons on basis of anti-vine blight linkage gene development and application of two molecular markers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Example 1, identification of resistance to vine blight of huapi shoot melon and xuelihong

[0077] The specific method is: select melon materials from the germplasm resource bank of the laboratory --- Huapishao melon, xuelihong, F1 offspring of Huapishaogua and xuelihong hybrid, inoculate with the pathogenic bacteria of muskmelon blight, and adopt the plant phenotype Symptoms determine resistance. Such as figure 1 .

[0078] 1. Inoculation of muskmelon blight pathogen:

[0079] 1) Cultivation of the pathogenic bacteria of variegated blight

[0080] Configuration of PDA medium: take 6.0 g of potato powder, 20.0 g of glucose, 20.0 g of agar powder, 2.0 g of ammonium dihydrogen phosphate, and 0.1 g of chloramphenicol. Dissolve the above components with distilled water and adjust the volume to 1 L, adjust the pH to 6 with NaOH or HCl, and sterilize at high temperature and high pressure for 20 minutes.

[0081] Activation of Pseudomonas spp.: Take out the strains preser...

Embodiment 2

[0097] Embodiment 2, muskmelon blight resistance genetic analysis and gene mapping:

[0098] Select melon materials from the germplasm resource bank of the laboratory --- Huapishao melon, xuelihong, F1 offspring of Huapishaogua and xuelihong hybridization and F2 population obtained by self-crossing of F1, and inoculate with pathogenic bacteria of muskmelon blight, The plant phenotype symptoms were used to determine the resistance, and the genetic analysis of disease resistance was carried out. Then, based on the high-throughput resequencing method, the disease resistance gene mapping is carried out, such as figure 2 .

[0099] Genetic analysis of resistance: the 219 individual plants of the F2 population were inoculated using the inoculation method for vine blight in Example 1, and the resistance was determined according to the phenotype. There were 165 disease-resistant plants and 54 susceptible plants. The number of disease-resistant plants: the number of melon-susceptibl...

Embodiment 3

[0111] Embodiment 3, the development and verification of marking CmGsbRS5 with KASP

[0112] The specific method is as follows: select melon materials from the germplasm resource bank of the laboratory --- Huapishao melon, xuelihong, F1 offspring of Huapishaogua and xuelihong hybridization and the F2 population obtained by selfing of F1, and first isolate the SNP Transformed into KASP markers, using primers to amplify KASP markers CmGsbRS5 and CmGsbRS6 to identify their genotypes and genetic segregation rules.

[0113] 1. Development of KASP markers CmGsbRS5 and CmGsbRS6:

[0114] 1. SNP information extraction:

[0115] According to the gene mapping results in Example 2, the SNP information closely linked with the disease resistance gene was extracted.

[0116] 2. Extract DNA

[0117] The genomic DNA of the F1 offspring of Huapishaogua, xuelihong, Huapishaogua and xuelihong hybrids was extracted, and the method was the same as in Example 2.

[0118] 3. PCR amplification

...

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Abstract

The invention belongs to the technical field of vegetable disease resistance molecular marker development and molecular marker assisted breeding, particularly relates to development and application ofa muskmelon vine blight resisting molecular marker, and provides a novel molecular marker and an assisted selection method for high-throughput screening identification and backcross breeding of vineblight resistance of muskmelons. The invention discloses two molecular markers which are developed on the basis of single nucleotide polymorphism linked with resistance to gummy stem blight, and are used for identifying vine blight resistance of the muskmelons. The invention also discloses application of the molecular markers, and the application is used for identification of vine blight resistinggermplasm of the muskmelons and auxiliary selected breeding of progenies of the muskmelons.

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 resistance to vine blight of melon, which provides a high-throughput screening identification and backcross breeding for resistance to vine blight of melon. A novel molecular marker and assisted selection method. Background technique [0002] Gummy stem blight (Gsb) is a fungal soil-borne disease caused by the infection of melon black rot small ball shell fungus (Dudymella bryoniae), which belongs to the eutrophic fungus and is one of the main diseases that harm melons. First, especially in Zhejiang Province and the southeast coastal areas, the conditions of high temperature and high humidity in protected cultivation are serious. The incidence rate in field fields can reach 20%-30%, and it can reach 80% in continuous cropping...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 杨景华张明方胡仲远
Owner ZHEJIANG UNIV
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