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SNP (single-nucleotide polymorphism) marker method for detecting cucumber Corynespora-cassiicola-resistant site

A technology of Corynesporium leaf spot and cucumber, which is applied in the field of SNP markers and identification of cucumber Corynesporium leaf spot resistance, can solve the problems that there are no relevant reports in the research of cucumber Corynesporium leaf spot assisted breeding, and achieve the method Simple, efficient and reliable results

Inactive Publication Date: 2016-05-25
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to domestic and foreign literature, so far, there has been no relevant report on SNP molecular marker-assisted breeding of cucumber Corynesporium leaf spot

Method used

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  • SNP (single-nucleotide polymorphism) marker method for detecting cucumber Corynespora-cassiicola-resistant site
  • SNP (single-nucleotide polymorphism) marker method for detecting cucumber Corynespora-cassiicola-resistant site
  • SNP (single-nucleotide polymorphism) marker method for detecting cucumber Corynespora-cassiicola-resistant site

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] The primers used for detecting the resistance of cucumber to Corynesporium leaf spot consist of the forward primer shown in SEQIDNo.1 and the reverse primer shown in SEQIDNo.2.

[0020] Forward primer F: 5'-accctgaccttccttatc-3' (SEQ ID No.1)

[0021] Reverse primer R: 5'-ggttgacagttgattaagc-3' (SEQ ID No.2)

[0022] The above primers are used to detect four SNP loci associated with resistance to Corynespora leaf spot of cucumber at bases 9718701, 9718752, 9718799 and 9718919 of the cucumber genome sequence Csa6M138620.1. The resistance to leaf spot disease was well typed, that is, the base forms at 9718701, 9718752, 9718799 and 9718919 were detected to express the differences in the resistance and susceptibility of cucumber to Corynespora leaf spot disease. The four SNP markers are located on the sixth chromosome of cucumber.

Embodiment 2

[0024] The SNP marker method for detecting the resistance locus of cucumber Corynesporium leaf spot, comprising the steps:

[0025] (1) extracting the cucumber genomic DNA of the tested sample;

[0026] (2) Using the cucumber genomic DNA as a template, and using the nucleotide sequences described in SEQIDNo.1 and SEQIDNo.2 as upstream and downstream primers, carry out PCR amplification, and put into the special thin-walled tube for PCR amplification: the Described cucumber genomic DNA 20ng, the nucleotide sequence 25ng described in SEQIDNo1 in the sequence listing, the nucleotide sequence 25ng described in SEQIDNo2 in the sequence listing, dNTP0.20mM, Mg 2+ 1.5mM, 1 times PCR buffer, 1.0 units of TaqDNA polymerase, add sterile double distilled water to 20μl, put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, the amplification conditions are: 94°C pre- Denaturation for 180 seconds, denaturation at 94°C for 60 seconds, annealing at...

Embodiment 3

[0032] Identification of Cucumber Germplasm Resources Corynesporium Leaf Spot Resistance

[0033] Cucumber germplasm resources XL6-1-2, 09L4, Q6, LX-11-1, P62-1-1, D507-1-1, Q5, JY118, etc. were selected from the breeding room of Tianjin Kerun Cucumber Research Institute. The method in Example 2 was used to operate: after testing, the amplification products of XL6-1-2, 09L4, Q6, LX-11-1, and Q5 all had the nucleotide sequence described in SEQIDNo. Leaf spot disease resources; P62-1-1, D507-1-1 amplification products appear the nucleotide sequence described in SEQIDNo.4, which is the resource of Corynesporium leaf spot disease; JY118 amplification products appear SEQIDNo.3 and SEQIDNo. . The nucleotide sequence described in 4 is the resource of impoverished Corynesporium leaf spot disease.

[0034]

[0035]

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Abstract

The invention discloses an SNP (single-nucleotide polymorphism) marker method for detecting a cucumber Corynespora-cassiicola-resistant site. The method comprises the following steps: (1) extracting genome DNA (deoxyribonucleic acid) of a detected sample cucumber; (2) by using the DNA as a template and sequences disclosed as SEQ ID No.1 and SEQ ID No.2 as forward and reverse primers, carrying out PCR (polymerase chain reaction) amplification to obtain a 319bp nucleotide sequence; and (3) carrying out sequencing on the nucleotide sequence: if the nucleotide sequence is disclosed as SEQ ID No.3, the sample is disease-resistant, and if the nucleotide sequence is disclosed as SEQ ID No.4 or simultaneously has the sequences disclosed as SEQ ID No.3 and SEQ ID No.4, the sample is disease-infected. The method is simple, and has the advantages of high efficiency and reliable result. The molecular marker can be used for molecule-assisted selective breeding of cucumber Corynespora cassiicola resistance, and can lay a foundation for improving the disease resistance of the cucumber varieties and completing the cucumber molecular breeding technique platform by gene engineering.

Description

technical field [0001] The invention relates to the fields of biogenetic engineering technology and molecular biology, and provides a SNP marker of cucumber corynesporium leaf spot resistance and an identification method thereof. Can be used for early molecular-assisted selection of cucumber Corynesporium leaf spot resistance to improve breeding efficiency. Background technique [0002] Cucumber (Cucumis Sativus L.) is an annual vine herbaceous plant of Cucurbitaceae (Cucurbitaceae) and the genus Cucumis (Cucumis). Cucumber target leaf spot [Cucumbertargetleafspot], also known as "brown spot", "target spot", "yellow spot disease", is a worldwide distribution of fungal diseases, serious outbreaks in my country in recent years, is recognized A disease that harms cucumber production is second only to downy mildew, and has become a key disease that restricts cucumber production. Corynesporium leaf spot of cucumber occurs in both greenhouses and open fields, and has a tendency t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 王惠哲曹明明邓强李淑菊杨瑞环刘耕春
Owner TIANJIN RES INST OF VEGETABLE
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