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Molecular markers and primers for detecting soybean phytophthora root rot resistance gene, detection method and application of molecular markers and primers
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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
Active Publication Date: 2019-09-24
安阳市农业科学院
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Problems solved by technology
[0003] However, Phytophthora sojae is highly variable and resistance in soybean varieties can be easily overcome
Method used
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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] Polypropylenegel 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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Abstract
The invention belongs to the technical field of soybean molecular breeding, and particularly relates to molecular markers and primers for detecting a soybean phytophthoraroot rot resistance gene, a detection method and application of the molecular markers and primers. The primers are a primer pair composed of upstream and downstream primers of a molecular marker sat186 and a molecular marker satt683 in the sequence table. The invention provides the soybean phytophthoraroot rot resistance genes molecular marker sat186 and molecular marker satt683, the linkage distances are 5.1 cM and 9.3 cM respectively, and the disease resistance gene Rps1498 has disease resistance to an ultrastrong phytophthora sojae strain PsJS2. The molecular markers closely linked to the disease resistance gene can be used for performing marker-assisted selection on the soybean phytophthora root rot resistance gene to prolong the disease resistance of the variety. The soybean phytophthora root rot resistance gene molecular markers and the special primers thereof can play an important role in soybean disease resistance breeding.
Description
technical field [0001] The invention belongs to the technical field of soybean molecular breeding, and in particular relates to molecular markers, primers, detection methods and applications for detecting soybean Phytophthora root rot resistance genes. Background technique [0002] Phytophthora root rot is one of the devastating diseases that seriously affect soybean production caused by Phytophthora soybean. Since Phytophthora soybean was first reported in Northeast China in 1989, [2] Its harmful area continues to expand, and it also causes serious harm in the Yellow River, Huaihe River and Yangtze River Basin in my country. Studies at home and abroad have proved that cultivating and planting disease-resistant and tolerant soybean varieties is one of the most effective measures to control soybean Phytophthora root rot. A large number of soybean varieties and resources resistant to Phytophthora root rot have been screened in my country, and some of the resistant varieties ...
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