Molecular markers linked to cucumber target spot disease resistance and their special primers and applications

A cucumber target spot disease, molecular marker technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. The control effect is not ideal and other problems, so as to improve the breeding efficiency and shorten the breeding years.

Active Publication Date: 2019-12-10
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 target spot disease is not ideal, which not only increases production investment but also causes environmental safety hazards, and stubble grafting makes technical difficulties and labor costs increase

Method used

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  • Molecular markers linked to cucumber target spot disease resistance and their special primers and applications
  • Molecular markers linked to cucumber target spot disease resistance and their special primers and applications
  • Molecular markers linked to cucumber target spot disease resistance and their special primers and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: Acquisition of Cucumber Target Spot Disease Resistance Gene Cca

[0024] 1. Preliminary and fine mapping of cucumber target spot disease resistance gene Cca

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

[0026] A. Mapping research on resistance genes of cucumber target spot disease Parents and genetic groups:

[0027] WF2757 (female parent) and Xintaimici (male parent) were selected as resistant parents, and a F2:3 population of 150 strains and a large F2 population of more than 2000 strains were constructed. Among them, the male parent Xintaimici and the female parent WF2757 were purchased from the Beijing Crop Germplasm Bank.

[0028] B. Investigation of Disease Index of Cucumber Target Spot Disease 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 were rai...

Embodiment 2

[0059] Example 2: Acquisition and functional verification of dCAPs marker CcaSNP1 co-segregated with cucumber target spot disease resistance gene Cca

[0060] 1. Acquisition of dCAPs marker CcaSNP1

[0061] It can be seen from Example 1 that there is a SNP in the 1481st position of the Cca gene between the resistant and susceptible materials, the base in the resistant material is G, and the base in the susceptible material is T, using this SNP site, according to dCAPs Primer online design website http: / / helix.wustl.edu / dcaps / dcaps.html, develop and design dCAPs molecular markers closely linked to resistant / susceptible target spot disease, named CcaSNP1, its upstream and downstream primers are shown in the sequence list as SEQ ID Shown in NO:1 and SEQ ID NO:2.

[0062] Through the above primer pair (SEQ ID NO: 1 and SEQ ID NO: 2 in the sequence listing), PCR amplification is carried out with the genomic DNA of the parental material (the disease-resistant material WF2757 and th...

Embodiment 3

[0068] Example 3 Application of dCAPs molecular marker CcaSNP1 to identify target spot disease resistance of cucumber materials

[0069] 1. Materials to be tested: WF2757 and Jinyan No. 2 were used as parents to construct a F2 genetic population, and 100 individual plants were randomly selected for the following experiments. Among them, the male parent Jinyan No. 2 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. Extract genomic DNA from the seedlings of the material to be tested, use each genomic DNA of the material to be tested as a template, utilize upstream and downstream primers SEQ ID NO: 1 and SEQ ID NO: 2 to carry out PCR amplification, and perform MaeII endonuclease to the PCR product...

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Abstract

The invention discloses a corynespora cassiicola disease-resistant linkage molecular marker as well as a special primer and application thereof. The molecular maker is one of nucleotide sequences amplified out of the total DNA of a 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 can be used for identifying the existence of a target gene namely a corynespora cassiicola disease-resistant gene,Cca to realize indirect selection of disease-resistant plants; and the molecular marker cannot be influenced by other gene effects and environmental factors, so that the molecular marker can be used for early generation selection, the breeding time is shortened, the breeding efficiency is improved, and a technical support is provided for performing corynespora cassiicola disease-resistant breeding.

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 with a cucumber target spot disease resistance gene Cca 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] Corynesporium leaf spot of cucumber, also known as target spot, is a worldwide distribution disease. At present, it has become one of the important diseases that endanger the cultivation of cucumbers in open field and protected areas, especially in protected areas such as overwintering greenhouses, winter-spring greenhouses, and spring greenhouses. It mainly harms the leaves. The lesions are light brown at the beginning and then turn brownish green. In severe cases, the leaves die. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6895
Inventor 温常龙董从娟毛爱军于拴仓许勇
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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