Indel Molecular Marker of Wheat Leaf Rust Resistance Gene lr13 and Its Application
A leaf rust resistance gene and molecular marker technology, applied in the field of biogenetics, can solve the problems of easy loss of resistance and difficulty in the existence of disease resistance genes, and achieve the effect of accelerating the process and improving the selection efficiency.
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Embodiment 1
[0029] The invention provides a method for screening molecular markers of wheat leaf rust resistance, specifically as follows:
[0030] 1. Genetic analysis
[0031] 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.
[0032] Table 1 Leaf rust i...
Embodiment 2
[0042] In this example, the indel molecular marker Lseq102 was used to screen 33 materials known to contain or not contain Lr13. The specific steps are as follows:
[0043] 1. Using the CTAB method to extract DNA from leaf tissues of 33 wheat materials.
[0044] 2. Using the genomic DNA extracted in step 1 as a template, a primer pair consisting of a single-stranded DNA molecule (upstream primer) shown in sequence 1 and a single-stranded DNA molecule (downstream primer) shown in sequence 2 in table 3 is used for PCR amplification. increase.
[0045] PCR amplification reaction system (10μl system): template DNA (50ngμL -1 ) 2.0 μL, 2×PCR Mix 5.0 μL (GenStar company product), upstream primer 1 μL, downstream primer 1 μL, ddH 2 O 1.0 μL.
[0046] The reaction program of PCR amplification: 94°C for 5min; 35 cycles of 94°C for 30s, 56°C for 30s, and 72°C for 30s; 72°C for 5min.
[0047] Table 3 The upstream and downstream primer sequences of Indel molecular marker Lseq102
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