Molecular markers and primers for detecting soybean phytophthora root rot resistance gene, detection method and application of molecular markers and primers
A technology of anti-Phytophthora root rot and molecular markers, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc. sick effect
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Embodiment 1
[0027] The specific method for screening the disease-resistant gene Rps1498 using the primer sequence is as follows:
[0028] S1, extracting the genomic DNA of the soybean to be detected;
[0029] S2, using the genomic DNA of the soybean as a template, respectively carrying out PCR amplification with primers of the molecular marker satt186 and primers of the molecular marker satt683, the PCR reaction system is:
[0030] 15ul system: 0.75ul of upstream and downstream primers, 1.5ul of template DNA, 4.5ul of ddH2O, 7.5ul of PCR Mix (2×) (including Taq DNA polymerase, dNTP and all components required for PCR except DNA template and primers 2× concentrated solution);
[0031] PCR program: 94°C for 5min; 32 cycles: 94°C for 30s, 57°C for 30s, 72°C for 40s; 72°C for 5min.
[0032] Polypropylene gel electrophoresis was performed using the PCR product of soybeans to be tested. In this example, the DNA template of Andou 1498 was used as a control. If the displayed bands were consiste...
Embodiment 2
[0034] Acquisition of closely linked molecular markers sat186 and satt683 of Phytophthora root rot resistance gene
[0035] S1, with Andou 1498 as the male parent and Williams as the female parent. The soybean variety Andou 1498 was bred by the Anyang Academy of Agricultural Sciences. The soybean variety Williams was provided by Mr. Li Haichao from the Economics and Research Institute of the Henan Academy of Agricultural Sciences. F 2:3 The family was obtained by crossing Andou 1498 and Williams to obtain F1, and then the F2 produced by selfing was harvested from a single plant, and the resulting seeds were sown in divisions.
[0036] S2, F 2:3 The disease resistance identification was carried out on generations of families. The tested strains were Race1, Race3, Race4, NKI, USAR2, Ps41-1, PsMC1, PsJS2; 8 Phytophthora sojae strains were obtained from the Crop Resistance Center of Crop Research Institute, Chinese Academy of Agricultural Sciences. Provided by the Pest Identifica...
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