Molecular marker primer for rapidly and efficiently identifying rice salt tolerant gene SKC1 and application
A salt-tolerant gene and molecular marker technology, applied in the field of plant biology, can solve the problems such as the inability to completely avoid reading errors of electrophoresis bands, prolonged test time, and the existence of toxic reagents.
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Embodiment 1
[0027] Example 1: Establishment of a molecular marker for rapid and efficient identification of rice salt tolerance gene SKC1
[0028] (1) Primer design
[0029] We extracted the genome sequence of SKC1 from the rice 3K database, including 7 SKC1 salt-tolerant genotype rice varieties (Nona, HD86, Pokkali, IR42, Zhenshan 97, Minghui 63, Gui 630), SKC1 salt-sensitive genotype rice varieties 7 (Koshihikari, Nip, Shengdao 19, Huaidao 5, Guangluai 4, Suxiu 867, Runnong 11). Through multiple sequence alignment, it is found that there is a G-C base-specific difference site at 994bp after the initiation codon ATG, such as figure 1 shown. This site is consistent with the existing reporter molecular marker design site (Jiang Yiwei et al., 2018), has the specificity and representativeness of the SKC1 genotype, and can be used as a molecular marker development site.
[0030] The nucleotide sequence related to the detection of the salt-tolerant type (Nona type) site is as follows (SeqNo...
Embodiment 2
[0043] Example 2: Using SKC1-Allele molecular markers to analyze Shengdao 19, HD86 and BC carrying HD86 fragments with Shengdao 19 as the recurrent parent 4 f 2 Genotypes of segregating populations
[0044] (1) Extraction of genomic DNA from rice leaves
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