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Molecular marker of cucumber fusarium wilt anti-disease gene Foc-4 as well as special primer and application thereof

A cucumber fusarium wilt, molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of adding technical difficulties and labor costs, increasing production investment, and causing environmental safety hazards and other problems to achieve the effect of improving breeding efficiency and shortening breeding years.

Active Publication Date: 2014-06-25
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effect of chemical control of Fusarium wilt is not ideal, which not only increases production input but also causes environmental safety hazards. Although stubble grafting is often used for prevention and control, it increases technical difficulties and labor costs

Method used

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  • Molecular marker of cucumber fusarium wilt anti-disease gene Foc-4 as well as special primer and application thereof
  • Molecular marker of cucumber fusarium wilt anti-disease gene Foc-4 as well as special primer and application thereof
  • Molecular marker of cucumber fusarium wilt anti-disease gene Foc-4 as well as special primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Acquisition of Cucumber Fusarium Wilt Resistance Gene Foc-4

[0024] 1. Preliminary and fine mapping of the resistance gene Foc-4 of cucumber wilt physiological race 4

[0025] The specific positioning method includes the following steps:

[0026] A. Cucumber Fusarium wilt resistance gene mapping research Parents and genetic groups:

[0027] Select WF2757 (male parent) and Jinyan No. 2 (female parent) as anti-susceptible parents, and construct 90 RIL-F 8 strains, 130 strains of F 2:3 populations and more than 2000 strains of F 2 large groups. Among them, the male parent WF2757 and the female parent Jinyan No. 2 were purchased from the Beijing Crop Germplasm Bank.

[0028] B. Investigation of Disease Index of Cucumber Fusarium Wilt Resistance Gene Mapping Research:

[0029] The seeds of parents and each group were wrapped with gauze, soaked in warm soup, and then sowed in 50-hole hole trays after germination at a constant temperature of 28°C. Seedlings we...

Embodiment 2

[0059] Example 2: Acquisition and functional verification of dCAPs marker FWSNP1 co-segregated with cucumber wilt resistance gene Foc-4

[0060] 1. Acquisition of dCAPs marker FWSNP1

[0061] It can be seen from Example 1 that there is a SNP in the Foc-4 gene between the resistant and susceptible materials, the base in the resistant material is A, and the base in the susceptible material is G, using this SNP site, according to the online design of dCAPs primers The website http: / / helix.wustl.edu / dcaps / dcaps.html develops and designs a dCAPs molecular marker closely linked to resistance / susceptibility to Fusarium wilt, named FWSNP1, and its upstream and downstream primers are as shown in SEQ ID NO: 1 and Shown in SEQ ID NO:2.

[0062] Through the above primer pair (SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence table), the genomic DNA of the parental material (the disease-resistant material WF2757 and the susceptible material Jinyan No. 2) was used as a template for PCR amplifi...

Embodiment 3

[0068] Example 3 Identification of Fusarium Wilt Resistance of Cucumber Materials Using dCAPs Molecular Marker FWSNP1

[0069] 1. Materials to be tested: F 2 Genetic population, from which 100 individual plants were randomly selected for the following experiments. Among them, the male parent Changchun Mici and the female parent WF2757 were purchased from the Beijing Crop Germplasm Bank.

[0070] 2. Wrap the seeds of the above-mentioned materials to be tested with gauze, soak them in warm water, sow them in 50-hole trays after accelerating germination at a constant temperature of 28°C, and raise seedlings in an air-conditioned greenhouse. Genomic DNA is extracted from the seedlings of the material to be tested, and each genomic DNA of the material to be tested is used as a template, and the upstream and downstream primers SEQ ID NO: 1 and SEQ ID NO: 2 are used to perform PCR amplification, and the PCR product is subjected to SalI endolysis Enzyme digestion identification, the...

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Abstract

The invention discloses a molecular marker of a cucumber fusarium wilt anti-disease gene Foc-4 as well as a special primer and an application thereof. The molecular marker is one of nucleotide sequences amplified from total DNA of cucumber by using the following primers: SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence table, and SEQ ID NO: 3 and SEQ ID NO: 4 in the sequence table. The molecular marker is capable of identifying the existence of target gene of cucumber fusarium wilt anti-disease gene Foc-4 and realizing the indirect selection to the anti-disease plant, the molecular marker is free from the influence of other gene effects and environment factors, so that the molecular maker can be used for early generation selection, shortening breeding age limit and improving breeding efficiency.

Description

technical field [0001] The invention relates to a molecular marker in the technical field of genetic engineering, in particular to a molecular marker linked to the cucumber wilt disease resistance gene Foc-4 and its application in the selection of cucumber germplasm resources. Background technique [0002] Cucumber (Cucumis Sativus.L) is widely cultivated in the world, and China is the country with the largest cultivation area and the highest total output of cucumber in the world. [0003] Cucumber wilt (Fusarium Wilt), also known as wilt disease, vine cut disease, dead seedling disease, is caused by the infection of Fusarium oxysporum f.sp.cucumerinum Owen. Soil infection, invading from roots or root necks, and parasitic systemic diseases in vascular bundles (catheter type fusarium wilt), is one of the three major diseases that are difficult to control in cucumber production. and other major soil-borne diseases of cucumbers in protected areas, which can cause a loss of 25%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 温常龙毛爱军董从娟张海英王永勤王永健于拴仓许勇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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