Wheat molecular marker and application thereof in identifying wheat powdery mildew resistance
A technology of wheat powdery mildew and molecular markers, which can be applied to the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problems of frequent loss of resistance and variable pathogenic species
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Embodiment 1
[0065] Example 1. Obtaining the molecular marker IWB41105 of the powdery mildew resistance gene and its special primer set K-IWB41105 and Str-IWB41105.
[0066] Field phenotype identification and data analysis: F 1 Generation, continuous selfing to get F 8 Substituting groups of RILs, the F 8 A total of 244 families of RILs were planted in Zhengzhou and Beijing in 2015 and 2016 for field identification of adult plant resistance to powdery mildew. The international general SAS statistical software PROC CORR model was used to calculate the Pearson correlation coefficient of MDS, and the PROC ANOVA command was used for variance analysis. The results of correlation analysis and variance analysis showed that the correlation coefficients of the four environments were between 0.53-073, and the correlation was good. There are extremely significant differences among different genes in powdery mildew resistance, which clarifies the validity of phenotypic data.
[0067] Genetic map co...
Embodiment 2
[0079] Embodiment 2, establishment of genotype detection method
[0080] 1. Use the K-IWB41105 primer set of Example 1 to detect Zhou 8425B / Chinese Spring F 8 The genotypes of 244 families in the generation RIL population, and the F 1 generation as a comparison.
[0081] 1. Extract Zhou 8425B / Chinese Spring F 8 The genomic DNA of 244 families in the RIL population was generated to obtain a template solution with a DNA concentration of 100 ng / μl.
[0082] 2. Using the genomic DNA extracted in step 1 as a template, use the primer set K-IWB41105 for PCR amplification to obtain PCR amplification products, and use pure water to replace the template as a control.
[0083] 5μl system for PCR amplification of K-IWB41105 primer set: KASP mix 2.5μl, MgCl 2 0.04μl, FAM primer 134nM, HEX primer 134nM, common reverse primer 336nM, genomic DNA 100ng, with ddH 2 O supplemented to 5 μl. KASPmix (KBS-1016-002, KASP V4.0 2X Master mix 96 / 384, Std Rox) and MgCl 2 All are products of LGC....
Embodiment 3
[0117] Example 3. Using the K-IWB41105 primer set and the Str-IWB41105 primer set to detect the genotypes of 103 Zhou 8425B derivatives.
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