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Four kinds of single nucleotide polymorphism-based molecular markers of muskmelon blight resistance and their application

A single nucleotide polymorphism, molecular marker technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problem of molecular markers and disease resistance genes. High-throughput screening and identification of vine blight, etc., to achieve the effect of improving vine blight resistance, efficiency and effect

Active Publication Date: 2021-04-27
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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  • Four kinds of single nucleotide polymorphism-based molecular markers of muskmelon blight resistance and their application
  • Four kinds of single nucleotide polymorphism-based molecular markers of muskmelon blight resistance and their application
  • Four kinds of single nucleotide polymorphism-based molecular markers of muskmelon blight resistance and their application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090]Example 1 Example 1, Flower Peel Telting Piga and Snow Red Cit Characters Defense

[0091]Choose a melon material from the laboratory germplasm --- Flower tip tap, snow red and flower peeling melons and snow red hybrids F1 progeny, melon sarametic bacteria vaccination, using plant pattern symptoms to determine resistance . Such asfigure 1 .

[0092]First, melon valence sick pathogen inoculation:

[0093]1), cultivation of citroen diseases

[0094]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 dihydrogen phosphate 2.0 g, chloramphenicol 0.1 g. The above components were dissolved with distilled water to 1 L, and pH was transferred to 6, high temperature and high pressure sterilization for 20 min with NaOH or HCl.

[0095]Activation of the citroening bacteria: The strain preserved in glycerin from the ultra-low temperature refrigerator is taken out, and the strain is inoculated into the PDA medium on the ultra-net working table, and ...

Embodiment 2

[0111]Example 2, melon spande disease resistance genetic analysis and genetically positioning:

[0112]Choose a melon material from the laboratory germplasm - trumberry, snow red, flower peel tap melon and snow red hybrid F1 progeny and F1 self-employed F2 group, melon sarametic pathogen inoculation, The resistance of disease resistance is performed using the plants of phenotypic symptoms. The disease is then performed based on high-throughput relegation sequencing method, such asfigure 2 .

[0113]Resistant Genetic Analysis: Taking 219 monographs of the F2 population in Example 1, inoculation of the F2 population, according to the phenotype decision resistance, the F2 population has a separation of the disease of melon vine sickness. 165 strains of anti-illnesses, 54 susceptible plants, number of anti-illnesses: melon sensation plant = 3: 1, indicating that the anti-valence traits of the melon were controlled by dominant monologically.

[0114]Resistant gene positioning:

[0115]First, extract...

Embodiment 3

[0125]Example 3, development and verification with KASP tags

[0126]The specific approach is to select a melon material from the laboratory germplasm - trumberry, snow red, flower skin tip and snow red hybrid F1 generation and F1 self-employed F2 group, first put SNP Transformed into KASP tags, using primer KASP tags CMGSBRS1, CMGSBRS2, ​​CMGSBRS3, CMGSBRS4 amplification to identify its genotype and genetic separation law.

[0127]1, KASP mark CMGSBRS1, CMGSBRS2, ​​CMGSBRS3, CMGSBRS4 development:

[0128]First, SNP information extraction:

[0129]The SNP information is extracted with the resistance gene closely chain according to the second embodiment.

[0130]Second, extract DNA

[0131]Extract the tip of tap melon, snow red, flower peel tap melon and snow red hybrid F1 possession genomic DNA, and the method is in Example 2.

[0132]Third, PCR amplification

[0133]1), the reaction system

[0134]The primer sequence is as follows:

[0135]CMGSBRS1: Aliquot 1: Gaatctaccaaaagtatcagtagaaagag

[0136]Aliquot 2: Gaatc...

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Abstract

The invention discloses four kinds of molecular markers developed based on single nucleotide polymorphisms linked to resistance to vine blight for the identification of resistance to vine blight of melon. The molecular markers are CmGsbRS1, CmGsbRS2, CmGsbRS3, and CmGsbRS4 with melon as the species. any of the . The invention also discloses the application of the molecular marker, which is used for identifying the melon blight-resistant germplasm or the auxiliary selection breeding of its progeny. When screening muskmelon germplasms resistant to creeping blight, select the germplasm whose genotype is consistent with the genotype of the disease-resistant melon or the genotypes of the melon and xuelihong at the same time for breeding.

Description

Technical field[0001]The present invention belongs to the field of vegetable anti-disease molecules development and molecular marking auxiliary breeding technology, and it is specifically involved in a marker development and application of melon anti-cranium-skin disease, which provides for melon anti-valer 's patient high-throughput screening identification and back breeding A novel molecular marker and auxiliary selection method.Background technique[0002]Gummy Stem Blight, GSB is a fungal native disease, caused by Dudymella Bryoniae infected, belonging to the normal necrotic dead nutritional fungi, is the main disease of harming melon First, especially in Zhejiang and southeast coastal facilities, high temperature and high humidity conditions, the incidence of Daejens can reach 20% -30%, and the incidence of 80% of the greenhouse and the greenhouse is high, and the destruction of severe disease is often devastated.[0003]Domestic and abroad have made a series of progress in the det...

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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