Molecular marker closely linked with corn bacterial wilt resistance genes and primer and application thereof
A molecular marker and resistance gene technology, applied in the field of agricultural biology, can solve the problems of low selection efficiency and long breeding cycle, and achieve the effect of reducing labor land, shortening breeding cycle, and improving corn yield level.
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[0028] Example 1. Obtaining the nucleotide sequence shown in SEQ ID NO: 3 in the sequence table and the nucleotide sequence shown in SEQ ID NO: 4 in the sequence table of the primer pair dedicated for identification of bacterial wilt resistance traits
[0029] 1. Parental genome amplification
[0030] The parents used to create the corn recombinant inbred line (RIL) population are 1145 and 0908 selected by our company. 0908 is the female parent and is not resistant to bacterial wilt; 1145 is the male parent and highly resistant to bacterial wilt.
[0031] The genomic DNA of the parental leaves was extracted by the CTAB method, and the primers published by Qing Yang (Qing Yang, Zhi Li, et al. CACTA-like transposable element in ZmCCT attenuated photoperiod sensitivity and accelerated the postdomestication spread of maize.PNAS 2013,10.) Yes, it consists of the nucleotide sequence shown in SEQ ID NO: 5 in the sequence table and the nucleotide sequence shown in SEQ ID NO: 6 in the sequenc...
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[0038] Example 2. Correlation verification between the amplified product of QK1 and corn bacterial wilt resistance traits
[0039] Using high-resistance material 1145 and non-resistance material 0908 as parents, 96 F after the two crossed 2 Substitute materials are the test objects. The genomic DNA of corn endosperm was extracted by alkaline boiling method, and the specific steps were the same as in Example 1. The PCR amplification experiment was performed with QK1. The PCR reaction system is: the PCR reaction system is: 10X Buffer 2μ1 (containing Mg2+), dNTP 0.4μ1 (1OmM), the nucleotide sequence shown in SEQ ID NO: 3 in the sequence table and the nucleotide sequence shown in SEQ ID NO: 4 in the sequence table The nucleotide sequence is 2μ1 (5μM), the genomic DNA extracted from corn to be tested is 1μ1, the taq enzyme is 0.5μ1 (5U / μ1), the reaction volume is 20μ1, and a drop of mineral oil is added to cover. The PCR reaction program is: 94℃ 5min; 94℃ 60s, 53.5℃ 60s, 72℃ 60s, 35...
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