The SNP Loci of Wheat Leaf Rust Resistance Gene lr13 and Its Application
A leaf rust resistance gene and leaf rust technology, applied in the field of biogenetics, can solve problems such as changes and affect protein functions, and achieve the effects of improving selection efficiency, high throughput, and accelerating the process of disease resistance breeding.
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Embodiment 1
[0032] The invention provides a method for screening molecular markers of wheat leaf rust resistance, specifically as follows:
[0033] 1. Genetic analysis
[0034] F 2 Phenotypic identification of individual plants in the population, F 2 The population contained 3057 individuals, of which 2300 were resistant and 757 were susceptible. F 2 The population contained 851 individual plants, including 624 resistant individual plants and 227 susceptible individual plants. Use SPSS to carry out chi-square test (suitability test) to the phenotype results. When the expected ratio is 3:1, the chi-square values are 0.092 and 1.268, P>0.05 (Table 2), and the resistance of Liaochun 10 is obtained. Leaf rust traits are controlled by a dominant single gene. Liaochun 10 and F derived from RL4031(Lr13) 2 All 1395 individual plants in the population were resistant to the disease, and it was concluded that the leaf rust resistance gene in Liaochun 10 was Lr13.
[0035] Table 1 Leaf rust i...
Embodiment 2
[0044] In this example, KASP marker primer Lseq302 was used to screen 33 materials known to contain or not contain Lr13. The specific steps are as follows:
[0045] 1. Using the CTAB method to extract DNA from leaf tissues of 33 wheat materials.
[0046] 2. With the genomic DNA extracted in step 1 as a template, PCR amplification is carried out with a primer pair consisting of a single-stranded DNA molecule (upstream primer) and a reverse single-stranded DNA molecule (downstream primer) shown in the sequences FAM and HEX in Table 3 .
[0047] Table 3 The upstream and downstream primer sequences of KASP marker Lseq302
[0048]
[0049] Lseq302 KASP labeled primers were dissolved to make a primer mix (12 mM primer sequence FAM and HEX, 30 mM reverse primer sequence), the total volume of the 384 plate reaction system was 5 μL, and 2×KASP V4.0 Mastermix was 2.5 μL (product of LGC Company) ), the primer mix is 0.056μL, the DNA template is about 50-100ng, and ddH 2 O make up...
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