SNP (single nucleotide polymorphism) marker related to melon powdery mildew resistance and application of SNP marker

A technology for powdery mildew and melon, applied in the field of molecular biology, can solve the problem that chemical agents are difficult to ensure pollution-free production very effectively.

Active Publication Date: 2016-07-27
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although powdery mildew can be controlled chemically, it is difficult for chemical agents to effectively ensure pollu

Method used

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  • SNP (single nucleotide polymorphism) marker related to melon powdery mildew resistance and application of SNP marker
  • SNP (single nucleotide polymorphism) marker related to melon powdery mildew resistance and application of SNP marker
  • SNP (single nucleotide polymorphism) marker related to melon powdery mildew resistance and application of SNP marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, acquisition of SNPs related to powdery mildew resistance of melon

[0064] In the previous research, the team of the inventor of the present invention cloned the Pm-2F gene by using map-based cloning technology (see "The title of the invention is "Plant powdery mildew resistance-related protein Pm-2F and its encoding gene and application", the application publication number is The Chinese patent application of CN105198977A), the full-length sequence of Pm-2F gene is utilized F 2 Comparing the sequence of the recombinant single plant with the resistant parent, it was found that F 2 The mutation sites of the Pm-2F gene at 23bp and 45bp (relative to the start codon ATG of the gene, where A is +1 position) in the recombinant single plant were consistent with those of the susceptible parent, while all mutation sites at other positions were identical. Consistent with the disease-resistant parent (the phenotypic identification result of the single plant is susce...

Embodiment 2

[0065] Example 2, Design of CAPS marker and its application based on the key SNP site of powdery mildew resistance gene Pm-2F

[0066] 1. Design of CAPS markers based on key SNP loci of powdery mildew resistance gene Pm-2F

[0067] The present invention utilizes the SNP site of the Pm-2F gene obtained in Example 1 in the powdery mildew resistant material (corresponding to the 23rd of the nucleotide sequence shown in sequence 8 in the sequence table in the melon genome; the SNP site The nucleotide at the dot is C or G), and the primers are designed according to the cleavage site at the mutated base, and the CAPS molecular marker (named as CAPS2-HphI molecular marker) closely linked with resistance / sensitivity to powdery mildew is developed and designed, wherein positive The forward and reverse amplification primers are:

[0068] Pm-2F-CAPS2-HphI-F: 5'-CGAGTCTTTCTTCTTCCAAATATCC-3' (SEQ ID NO: 1);

[0069] Pm-2F-CAPS2-HphI-R: 5'-GAAAGATTCATAGGAGAACTCGTCC-3' (SEQ ID NO: 2).

[...

Embodiment 3

[0096] Example 3. Design of high-throughput KASP molecular marker primers and its application based on the key SNP loci of the powdery mildew resistance gene Pm-2F

[0097] 1. Design of high-throughput KASP molecular marker primers

[0098] The present invention utilizes the SNP site of the Pm-2F gene obtained in Example 1 in the resistant material (corresponding to the 23rd of the nucleotide sequence shown in sequence 8 in the sequence table in the melon genome; at the SNP site The nucleotides are C or G), and high-throughput KASP molecular markers are designed.

[0099] The sequence of marker primers used for high-throughput screening is as follows:

[0100] Pm-2F-F1: 5'-CTATGGCAGAATCAATTCTGTTCA C -3';

[0101] Pm-2F-F2: 5'-CAATGGCAGAATCAATTCTGTTCA G -3';

[0102] Pm-2F-R: 5'-GTGGGAAAGAACCCAATTTTGTTGC-3'.

[0103] For the SNP site, the 3' ends of the upstream primers Pm-2F-F1 and Pm-2F-F2 are allelic variant bases (C or G in the bold underlined part), and the Pm-2F-F1 ...

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Abstract

The invention discloses an SNP (single nucleotide polymorphism) marker related to melon powdery mildew resistance and an application of the SNP marker. The application provided by the invention is the application of SNP at a following SNP site in a melon genome or a substance for detecting SNP of the following SNP site in the melon genome in identification or assistant identification of the melon powdery mildew resistance; the SNP site in the melon genome corresponds to the 23rd site in a nucleotide sequence shown in sequence 8 in a sequence table; nucleotide at the SNP site is C or G. The SNP marker provides technical support for selective breeding and molecular breeding of melon powdery mildew resistant varieties, and has significant application value in selective breeding of melon powdery mildew resistant varieties.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a SNP marker related to powdery mildew resistance of melon and its application. Background technique [0002] Melon (Cucumismelo L.) is an important economic crop of Cucurbitaceae Cucumber. Powdery mildew is a worldwide disease that seriously endangers the production of muskmelon. It can lead to the decline of plant photosynthetic ability, premature aging and even death, seriously affecting the yield and quality of muskmelon. In recent years, with the scale of production and the improvement of commercialization, powdery mildew has spread rapidly and has become a major obstacle to the green production of melons such as melons at home and abroad. Although powdery mildew can be controlled chemically, it is difficult for chemical agents to effectively ensure pollution-free production because there are many physiological races of melon powdery mildew, rapid differentiation, and easy to...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11A01H1/02
CPCA01H1/02C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 许勇张春秋张海英宫国义郭绍贵
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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