水稻悬浮细胞中稻瘟菌激发子诱导性受体类似激酶cDNA的克隆及特征分析  

CLONING AND CHARACTERIZATION OF AN ELICITOR INDUCIBLE RECEPTOR LIKE KINASE cDNA FROM RICE SUSPENSION CELLS

作　　者：范军[1] 彭友良[1] Qun Zhu ChrisLamb 

机构地区：[1]中国农业大学植物病理学系,北京100094 [2]The Salk Institute for Biological Studies

出　　处：《植物病理学报》1999年第3期235-241,共7页Acta Phytopathologica Sinica

基　　金：洛克非勒基金会水稻生物技术项目;国家杰出青年科学基金

摘　　要：采用改进的差异显示方法由稻瘟菌激发子处理的水稻悬浮培养细胞中得到７个诱导表达的蛋白激酶 ｃＤＮＡ 片段。为获得这些蛋白激酶的编码全长序列，构建了稻瘟菌激发子处理的水稻细胞ｃＤＮＡ 文库。以上述一个 ｃＤＮＡ 片段作为探针筛选文库，得到一个含有完整阅读框的 ｃＤＮＡ 克隆ＯｓＥＰＫ１。分析ＯｓＥＰＫ１的推定氨基酸序列的结果表明：它由Ｎ 端信号肽、胞外区域、跨膜结构和胞内蛋白激酶结构域等几部分组成，可能为一受体类似蛋白激酶。对比 ＯｓＥＰＫ１与其它植物受体类似蛋白激酶在胞外区域的相似性，结果显示ＯｓＥＰＫ１与小麦叶锈激酶家族同源性较高，推测它们可能属于同一类植物受体类似蛋白激酶。此外，ｎｏｒｔｈｅｒｎ 杂交显示，ＯｓＥＰＫ１可由稻瘟菌激发子迅速瞬时诱导，因而它的诱导可能也是水稻对稻瘟菌侵染做出的一种早期防卫反应。Using a modified protocol of mRNA differential display, the authors identified 7 induced cDNA fragments of protein kinase transcripts in rice suspension cells treated with blast fungus derived elicitor. A cDNA library of the elicited rice suspension cells was constructed to screen for the full coding region of these protein kinase transcripts. From the library, one cDNA clone named OsEPK1 was obtained with intact open reading frame. Computer analysis of its deduced amino acid sequence showed that it consists of a cleavable N terminal signal peptide, a large extracellular domain, a transmembrane domain and a cytoplasmic protein kinase domain, indicating that OsEPK1 might be a receptor like kinase. By comparing the putative extracellular domain of OsEPK1 with that of other plant receptor like kinases,we find that OsEPK1 is homologous to leaf rust kinases, which were recently identified in wheat, and hence speculate that they belong to the same type of plant receptor like kinases. In addition, result of northern blotting showed that OsEPK1 was transiently induced at transcription level in rice cells after treated with the elicitor, suggesting that induction of OsEPK1 might be one of early defence responses of rice to blast fungus infection.

关 键 词：水稻 激发子 基因表达 克隆 稻瘟苗 CDNA 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治] Q78[农业科学—植物保护]