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Marking method of CAPS related to cucumber sterility gen gene and application

A male sterility gene and marker technology is applied in the field of CAPS molecular markers for identifying cucumber fertility, and can solve problems such as background detection of cucumber fertility

Inactive Publication Date: 2019-02-22
TIANJIN RES INST OF VEGETABLE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purpose of the invention is to solve the problem of cucumber fertility background detection

Method used

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  • Marking method of CAPS related to cucumber sterility gen gene and application

Examples

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Embodiment 1

[0060] This example shoots to identify the cucumber male sterility gene Csa3M006660 wild type, heterozygous mutant, and homozygous mutant specific CAPS marker establishment

[0061] (1) Primer design:

[0062] Fine mapping of the male sterility gene ms-3 in cucumber (Cucumis sativus L.) mutants by Tianjin Cucumber Research Institute (Han, Y., et al. (2017). "Fine mapping of a male sterility gene ms-3 Ina novel cucumber (Cucumis sativus L.) mutant. "Theor Appl Genet.) research shows that if the 1258th base of Csa3M006660 gene is T, then the plant is sterile. Using Primer premier 6.0 software to design the upstream and downstream primers JD-CAPS2.0 containing the site on both sides of the 1bp mutation region.

[0063] Upstream primer JD-CAPS2.0-F: 5’-AGGAAGAGAAGTGAAGCAA-3’

[0064] Downstream primer JD-CAPS2.0-R: 5’-CCGTAACGTAATATGTGTCT-3’

[0065] (2) Theoretical analysis of amplification and restriction fragments:

[0066] Wild-type genomic DNA can amplify a 618 bp fragment (listed in ...

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Abstract

The invention discloses a specific CAPS marker for identifying a wild type, a heterozygous mutant and a homozygous mutant of a cucumber male sterility gene Csa3M006660. The specific CAPS marker is characterized in that the specific CAPS marker comprises a marker JD-CsaMS-CAPS2.0 for detecting a wild type locus, and a the nucleotide sequence of the marker JD-CsaMS-CAPS2.0 is shown as SEQ ID NO:1; and wild type genome DNA is shown as SEQ ID NO:2, and a mutant genome is shown as SEQ ID NO:3. By designing a method of combination of primer amplification, enzyme digestion and agarose gel electrophoresis, two characters of male sterility and male fertility in a cucumber 'YL-5' strain can be efficiently screened at the seedling stage. A method has the characteristics of short period, low cost andhigh efficiency, identifying can be conducted during on crossbreeding and, backcross segregation of colonies and the like, and a heterozygous genotype and a homozygous genotype are distinguished accurately, and the breeding efficiency is improved.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and particularly relates to a CAPS molecular marker method and application for identifying cucumber fertility. Background technique [0002] In the spring of 2012, we discovered a cucumber male sterile mutant in a high-generation inbred line at Tianjin Wuqing Experimental Site. After crossing, selfing and backcrossing methods, we identified that the sterile trait is a nuclear male controlled by a single gene. Infertility, and the infertility trait is recessive relative to the fertility trait. Through the construction of F2 mixed pool, re-sequencing technology and the method of constructing genetic map were used to finely map the sterility gene, and the final result was determined to be Csa3M006660 (Han, Y., et al. 2017.). [0003] The existing patent CN107058552A is an identification method based on the level of sequencing, by extracting mixed pool genomic DNA, combined with resequencing technology, sequen...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 韩毅科杜胜利宋卓然魏爱民刘楠
Owner TIANJIN RES INST OF VEGETABLE
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