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Piz-t是一个具有广谱稻瘟病抗性的基因,对于选育稻瘟病抗性品种具有重要的应用价值,但其所在位点多个不同的等位基因限制了对它的筛选及利用。本研究通过将Piz-t不同等位基因及其上下游测序,比对鉴定到一个Piz-t特异性SNP位点(Piz-t为A,其余均为G)。在两对交叉引物PCR(PCR with confronting two-pair primers,PCR-CTPP)方法的基础上加以改进开发了一套鉴定该SNP的标记,利用该标记及待检测水稻品种基因组DNA进行PCR扩增,通过不同PCR产物条带大小将Piz-t与其它等位基因区分开。该方法简便快速、成本低廉,可广泛应用于分子标记辅助选择育种或者水稻种质资源Piz-t基因型鉴定。
Piz-t is a broad-spectrum rice blast resistance gene, which has important application value for breeding rice blast-resistant varieties. However, many different alleles at its locus limit its selection and utilization. In this study, a Piz-t specific SNP was identified by sequencing different alleles of Piz-t and its upstream and downstream sequences (P of Piz-t was A, and the rest were G). Based on the PCR-CTPP method, a set of markers for identifying the SNP was developed. Using the marker and the genomic DNA of the rice varieties to be tested for PCR amplification, Piz-t is distinguished from other alleles by band sizes of different PCR products. The method is simple, rapid and low in cost, and can be widely applied to molecular marker-assisted selection breeding or Piz-t genotyping of rice germplasm resources.