Novel cucumber SNP molecular marker

A molecular marker, cucumber technology, applied in the biological field, can solve the problems of complex, long genetic distance between disease resistance genes and molecular markers

Inactive Publication Date: 2015-10-21
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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Problems solved by technology

Researchers have used molecular marker techniques such as AFLP and SSR to initially locate disease-resistant genes, but the genetic distance between disease-resistant genes and molecular markers is generally far away, indicating that the molecular mechanism of resistance to cucumber powdery mildew is very complicated. The problem needs to be studied and explored. Fully excavating and utilizing these resistance genes will enrich the resistance gene resources in cucumber powdery mildew resistance breeding and provide an important basis for introducing excellent resistance genes in cucumber powdery mildew resistance breeding.

Method used

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Embodiment

[0024] The young leaves of the material at the stage of two leaves and one heart were taken, and the genomic DNA was extracted by the improved SDS method, diluted to 60 ng / μL, and the concentration was detected by 1% agarose electrophoresis. The enzyme digestion prediction software SLAF_Predict system was used to analyze the cucumber genome, and the enzyme digestion scheme, gel cutting range and sequencing volume were determined according to its GC content, repeat sequence and gene characteristics, so as to ensure the density and uniformity of molecular markers.

[0025]Take 500ng of genomic DNA, add 0.6U Mse I (NEB, Hitchin, Herts, UK), T4 DNA ligase (NEB), ATP (NEB) and Mse I linker (NEB), react at 37°C for 15h, anneal at 65°C 1h, then add restriction endonucleases Hae III and Bfa I and react at 37°C for 3h. After the reaction, the digested product was purified with Quick Spin column (Qiagen), and redissolved with 33 μL EB. Use the recovered digested product as a template f...

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Abstract

The invention discloses a novel cucumber SNP molecular marker. The mark number of the novel cucumber SNP molecular marker is SLAF7695. According to the novel cucumber SNP molecular marker, the simplified genome depth sequencing technology is utilized, 73100 SLAF labels are obtained totally, 5355 polymorphic markers of three types including SNP, EPSNP and INDEL are obtained through polymorphic analysis, and 140 different markers closely related to cucumber powdery mildew resistance are obtained and mainly concentrated on the first chromosome and the sixth chromosome. Two candidate areas related to powdery mildew resistance characters are successfully located, seven special markers closely linked with cucumber powdery mildew resistance are obtained, 28 genes and related annotation information are obtained through gene annotation, and five genes are related to the functions of defense response, harm response, toxin catabolism, cell stress response and the like. The novel cucumber SNP molecular marker provides a theoretical foundation for exploration of cucumber powdery mildew resisting mechanism and molecular marker assisted selective breeding.

Description

technical field [0001] The invention relates to the field of biology, in particular to a novel cucumber SNP molecular marker. Background technique [0002] Cucumber powdery mildew is a widespread worldwide disease and one of the main cucumber leaf diseases. Its incubation period is short, its prevalence is strong, and it occurs every year, which seriously affects the yield and quality of cucumber. At present, the main control measures for cucumber powdery mildew are chemical control, but the pesticide residues in the product are a safety issue that cannot be ignored, especially after using facility cultivation, the problem of continuous cropping obstacles in cucumber production is becoming more and more serious. Therefore, breeding disease-resistant varieties is still an effective way to fundamentally control powdery mildew. [0003] The emergence of molecular marker technology has made the accurate detection of genes and the accurate transfer of target genes a reality. R...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 张鹏周胜军朱育强陈丽萍
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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