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Molecular marker-assisted selection primer and method for wilt resistance and gynoecious line of cucumis melon

A molecular marker-assisted, melon fusarium wilt technology, applied in the field of molecular genetics, can solve the problems of high cost of seed production, high labor intensity, and increased difficulty in emasculation, and achieve the effects of improving efficiency, reducing workload and saving costs.

Inactive Publication Date: 2014-06-18
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only the labor intensity is high, the cost of seed production is high, but also the purity of seeds is difficult to guarantee
In addition, thin-skinned muskmelon is the main cultivar of muskmelon in my country, and its fully flowered stamens are relatively small, which makes it more difficult to detassell, and it is even more difficult to guarantee the purity of the seeds.

Method used

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  • Molecular marker-assisted selection primer and method for wilt resistance and gynoecious line of cucumis melon
  • Molecular marker-assisted selection primer and method for wilt resistance and gynoecious line of cucumis melon
  • Molecular marker-assisted selection primer and method for wilt resistance and gynoecious line of cucumis melon

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Experimental program
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Effect test

Embodiment 1

[0066] Tested melon and its DNA extraction

[0067] (1) Analysis of the test melon and its resistance to Fusarium wilt

[0068] (1) Tested melon materials: WI998 and MR-1, F1 generation, F2 generation, disease-resistant control materials: 3-1-3, PI1446928, PI1446931, PI1446930, PI1446929, PI618848, PI508448, Taitian 2-2-4 , Taitian 3-1, No.30, susceptible control materials: Tianshuai, BT3, MRIL7-165, No.26, Taitian 1-5-1, TN, 6-1-4-4, S3, M -021, 3-2-2.

[0069] (2) Fusarium wilt resistance analysis at the seedling stage: The disease resistance was identified by inoculating the fusarium wilt pathogen by the method of root wounding at the seedling stage. When the test melon seedling grew to 1 true leaf, it was dug out, washed with water, and placed in spores. Suspension (the number of spores in the spore suspension is generally 5.5 × 10 5 Soak for 15min in pcs / mL), then replanted back to the nutrient cup for culture, culture conditions: temperature is 28°C, light duration is...

Embodiment 2

[0074] Target gene molecular marker screening

[0075] (1) Primer design for molecular marker screening of the target gene: traditional breeding methods were adopted, and the all-female Fusarium wilt-susceptible material WI998 was used as the female parent, and the androgenetic Fusarium wilt-resistant material MR-1 was used as the male parent. According to the difference in the 56th amino acid between the transcribed protein sequences of the A and a genes, that is, there is a single gene mutation between the 6045th base of the DNA sequence of the A gene and the corresponding site of the a gene, and this base site is found. CAPS primers were designed for the Alu I restriction site. Between ORF2 and ORF3 of the G gene, a transposon named Gyno-hAT was inserted into the ORF31.3kb region, so that the G gene could not be expressed normally and then inactivated. Therefore, SCAR primers were designed on the G and g genes respectively to distinguish between dominant and recessive gene...

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Abstract

The invention discloses a molecular marker-assisted selection primer and method for the wilt resistance and gynoecious line of cucumis melon. According to the method, CAPS (cleaved amplified polymorphic sequence) primers A-as and A-s are designed according to single-gene mutation between a 6045th basic group on the DNA (deoxyribonucleic acid) sequence of a gene A of the cucumis melon and a corresponding locus of a gene a, and an SCAR (sequence characterized amplified region) primer pair G-as and G-s and an SCAR primer pair g-as and g-s are respectively designed according to genes G and g of the cucumis melon and are used for differentiating and labeling sex-determining genes of the cucumis melon; and meanwhile, a comparison result of genes MR-1 and WI998Fom-2 at an SCOP functional domain shows that only a 481th basic group can be developed as a CAPS-marked site, and a primer pair F-s and F-as is designed according to the comparison result. The polymorphism that the digestion is carried out after amplification by virtue of a PCR (Polymerase Chain Reaction) can be applied to the molecular marker-assisted breeding of the wilt resistance and gynoecious line of the cucumis melon. The primer and the method are used for carrying out assisted breeding and have the advantages of definite target selection, cost conservation and the like.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a primer and a method for resistance to fusarium wilt of muskmelon and molecular marker-assisted selection of all female lines. Background technique [0002] Melon (Cucumis melon L.) is an important economic crop in my country. At present, androgynous is the main variety in commercial cultivation. In hybrid seed production, the complete corolla must be removed at the bud stage and emasculated. During this period, operation procedures such as bagging, pollination, marking, and seed purity identification increase labor intensity, high cost of seed production, and difficulty in ensuring seed purity. The stamens of the thin-skinned muskmelon are smaller than those of the thick-skinned muskmelon, and the purity is more difficult to guarantee. Moreover, the thin-skinned muskmelon is the main cultivar of muskmelon in my country. [0003] In the traditional melon seed production proc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 栾非时高鹏王学征王凤娇马鸿艳朱子成
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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