Two Molecular Markers Developed Based on the Linkage Genes of Resistance to Dwarf Blight and Their Applications

A technology of molecular markers and blight disease, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. The problem of long genetic distance of the disease gene can be solved, and the effect of improving the resistance to the blight and improving the efficiency and effect can be achieved.

Active Publication Date: 2021-06-01
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 Developed Based on the Linkage Genes of Resistance to Dwarf Blight and Their Applications
  • Two Molecular Markers Developed Based on the Linkage Genes of Resistance to Dwarf Blight and Their Applications
  • Two Molecular Markers Developed Based on the Linkage Genes of Resistance to Dwarf Blight and Their Applications

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 the hybrid of Huapishaogua, xuelihong, Huapishaogua and xuelihong was extracted, and the method was the same as in Example 2.

[0118] 3. PCR amplificat...

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Abstract

The invention belongs to the technical field of vegetable disease-resistance molecular marker development and molecular marker-assisted breeding, in particular to the development and application of a melon-resistant vine blight molecular marker, and provides high-throughput screening identification and backcross breeding for muskmelon resistance to vine blight. A novel molecular marker and assisted selection method. The invention discloses two molecular markers developed based on the linked single nucleotide polymorphism of resistance to vine blight and used for identification of vine blight resistance of melon . The invention also discloses the use of the above molecular marker: the auxiliary selection breeding for identifying the melon blight resistant germplasm or its progeny.

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 Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 杨景华张明方胡仲远
Owner ZHEJIANG UNIV
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