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Indel molecular marker co-segregating with cucumber ZymV resistance gene

A molecular marker and resistance gene technology, applied in the field of genetic engineering, can solve the problems of unsuitable large-scale population screening, the development of unfavorable molecular marker-assisted breeding, and the long genetic distance of linked markers, so as to achieve the effect of accelerating the process and large differences.

Inactive Publication Date: 2019-09-27
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The above studies show that ZYMV resistance in cucumber is controlled by a single recessive gene. Some linked markers obtained so far have a long genetic distance or are not suitable for large-scale population screening, which is not conducive to the development of molecular marker-assisted breeding.

Method used

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  • Indel molecular marker co-segregating with cucumber ZymV resistance gene
  • Indel molecular marker co-segregating with cucumber ZymV resistance gene
  • Indel molecular marker co-segregating with cucumber ZymV resistance gene

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Experimental program
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Embodiment

[0023] 1. Development and identification of InDel markers based on cucumber ZYMV resistance candidate genes

[0024] 1. Verification of multiple resistant and sensitive inbred lines

[0025] The sequence shown in SEQ ID NO.5 is obtained by sequencing the candidate gene in multiple cucumber disease-resistant inbred lines, and the sequence shown in SEQ ID NO.6 is obtained by sequencing the candidate gene in multiple cucumber disease-resistant inbred lines; both There is a 37bp insertion / deletion mutation between them. According to this mutation, the present invention uses Primer 5.0 primer design software to design forward and reverse primers on both sides of the mutation. The upstream primer is shown in SEQ ID NO.3, the downstream primer is shown in SEQ ID NO.4, the amplified band of the resistant material is 211bp, and the amplified band of the susceptible material is 174bp, through polyacrylamide gel Electrophoresis can quickly differentiate the genotypes of the materials us...

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Abstract

The invention relates to an InDel molecular marker co-segregated from a ZYMV resistance gene. The InDel molecular marker named InDel-zym1 and is composed of a nucleotide sequence fragment shown in SEQ ID NO. 1 and a nucleotide sequence fragment shown in SEQ ID NO. 2, wherein the nucleotide sequence fragment shown in SEQ ID NO. 1 is co-segregated from a resistance gene; the nucleotide sequence fragment shown in SEQ ID NO. 2 is co-segregated from a disease-susceptible gene. The InDel molecular marker provided by the invention has high stability and can be easily and rapidly applied to the auxiliary screening of ZYMV resistant individual plants in a cucumber seedling stage at high throughput; according to SEQ ID NO. 5 and SEQ ID No. 6, multiple pairs of co-dominant InDel markers can also be designed as molecular markers for ZYMV resistance screening on account of difference in DNA length of resistance candidate genes caused by insertion / deletion mutations. The InDel marker provided by the invention lays a foundation for ZYMV resistance molecular marker-assisted breeding, and greatly accelerates progress of breeding of ZYMV resistance molecules of cucumber.

Description

technical field [0001] The invention relates to genetic engineering technology, in particular to InDel molecular marker co-separated with cucumber ZYMV resistance gene. Background technique [0002] Cucumber (Cucumis sativus L.) is an annual herbaceous climbing plant in Cucurbitaceae. Originating in the northern part of India at the southern foot of the Himalayas, my country has a cultivation history of more than 2,000 years and is the secondary origin center of cucumbers. As one of the ten most important vegetable crops in the world, cucumber is also one of the main vegetable crops in my country. As an important vegetable crop, it has always been valued by breeders. [0003] In recent years, ZYMV (Zucchini Yellow Mosaic Virus, Cucumber Yellow Mosaic Virus) has become more and more harmful to cucumbers. After cucumbers are infected with ZYMV, the leaves appear yellowing, mosaic, and there are tumor-like protrusions on the surface of the leaves. When the disease is severe, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
Inventor 杨义何欢乐潘俊松蔡润朱文莹潘健杜慧
Owner SHANGHAI JIAOTONG UNIV
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