摘要:
Pik位点包含Pi-k、Pi-km、Pi-kp、Pi1、Pi-kh等抗稻瘟病等位基因，在我国水稻抗病育种中具有重要应用价值。本研究对229份水稻品种及重要育种材料的Pik位点进行基因型鉴定。利用K/N-单元型分子标记对水稻材料Pik位点进行多态性分析，鉴定了80份材料为K1K2功能性基因型。进一步通过Pi-k、Pi-kp和Pi1功能分子标记检测，结合功能多态位点序列分析，从80份K1K2型材料中，鉴定了黑稻、京虎B为Pi-k基因型；软米谷为Pi-kp基因型；恩恢99-64为Pi1基因型。研究还发现，高配合力强优恢复系闽恢3301的Pik位点为功能性K1K2基因型，其K1K2片段的序列与已克隆Pi-k、Pi-km、Pi-kp、Pi1、Pi-kh等位基因的序列存在差异，表明闽恢3301的Pik位点可能存在一个新等位基因。本研究明确了229份水稻材料Pik位点的K/N-基因型，结果可为水稻抗稻瘟病育种和品种合理布局提供参考。
Abstract:
The rice Pik locus that contains multiple blast resistance genes, such as Pi-k, Pi-km, Pi-kp, Pi1 and Pi-kh, has great utilization value in rice breeding for blast resistance in China. In the present study, we investigated the genotypes at the Pik locus in 229 rice cultivars and important breeding materials. Based on PCR amplification using K- and N-type specific primer sets, 80 accessions were identified harboring the K1K2 haplotypes. The 80 K1K2 accessions were further accessed for the presence of Pi-k, Pi-kp and Pi1 using allele-specific markers in combination with sequencing identification of allele-specific SNPs. Heidao and Jinghu B were identified harboring the Pi-k resistance allele, and Ruanmigu and Enhui 99-64 were identified harboring Pi-kp and Pi1, respectively. In addition, an elite indica restorer line Minhui 3301 was identified harboring the K1K2 haplotypes. Allele-specific marker analysis and sequencing data revealed that the sequence of Minhui 3301 at Pik locus differs from the sequences of Pi-k, Pi-km, Pi-kp, Pi1 and Pi-kh, suggesting that Minhui 3301 might possess a new allelic gene at the Pik locus. In summary, we validated the distribution of the K1K2 haplotypes at the Pik loucs in 229 rice accessions. Our results provide a wealthy of information for developing and deploying resistant cultivars to control rice blast disease.
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