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.
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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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