水稻不育系系谱抗稻瘟病基因同源序列的克隆与序列分析

Cloning and sequences analyzing of resistance genes from rice blast sterile line lineage

根据已知NBS-LRR类抗病基因结构中氨基酸的保守区域设计简并引物,对水稻不育系系谱NBS-LRR类抗病基因同源序列进行克隆、测序和聚类。同源序列克隆与分析表明,7个水稻不育系系谱亲本中共获得14个阳性克隆,其中11个含有NBS-LRR类抗病基因所特有的保守氨基酸结构域KinaseⅠ、KinaseⅡ、KinaseⅢ及跨膜区域。这些片段间同源性最高达98.3%,最低仅为29.7%;11个氨基酸片段与抗病基因Xal(AB002266)的同源性较高,42%-45%的氨基酸序列相同,59%-64%的氨基酸序列相似,且与水稻其他NBS-LRR类抗性蛋白相似性很高。通过聚类分析,可以将其分为3类,分别记为G1、G2、G3,并发现抗源谷农13所含有的G3类基因,通过天谷、福伊传给后代谷丰、全丰。

According to the known conserved domain of the amino acid in the NBS-LRR structure genes, we could design the degenerate primer and clone, sequence and cluster the resistance gene analogs （RGAs） by PCR in order to study the passing rules of resistance genes and analyze the comparability of blast phenotype and RGAs, offering the scientific reference for utility and clone of rice blast genes. The cloning and analysis of the homologous sequence of resistant genes of the rice sterial line lineage showed that 14 masculine clones had been obtained from 7 rice sterial line lineage. And there were only 11 NBS-LRR resistant genes which had particular conservative domain, Kinase Ⅰ , Kinase Ⅱ , Kinase Ⅲ and transmembrance domain. In the amino acid segments of the NBS-LRR type, the maximal homologous quality was 98.3%, the minimum was 29.7%. These 11 NBS-LRR type sequences were highly homologic to resistant gene Xal （AB002266）, which had 42% -45% identical amino acids and 59% -64% similar amino acids by the BLAST in NCBI. These NBS-LRR type resistant sequences had been separated into three types, which were named G1, G2, G3 by clustering analysis. And it showed that the G3 type genes of Gunongl3 had been passed to generation of Gufeng and Quangfeng through the generation Tiangu and Fuyi.