Cloning of genes encoding nonhost hypersensitive response-inducing elicitors from Phytophthora boehmeriae  

作　　者：LI Jun ZHANG HaiFeng ZHANG ZhengGuang WANG YuanChao ZHENG XiaoBo 

机构地区：[1]Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, China [2]Suzhou Plant Protection and Quarantine Station, Suzhou 215006, China

出　　处：《Chinese Science Bulletin》2007年第2期231-237,共7页

基　　金：the Nation Natural Science Foundation of China (Grant No. 30300228);the National Research Foundation for the Doctoral Program of Higher Education of China (Grant No. 20020307035);the Student Research Training of Nanjing Agricultural University (Grant No. 0402A04)

摘　　要：We have devised a high-throughput functional cloning method to isolate cDNAs from Phytophthora boehmeriae of which the products elicit a hypersensitive response (HR) in tobacco. The cDNAs were cloned into a binary potato virus X (PVX)-based expression vector and transformed into Agrobacterium tumefeciens (Mog101). 4100 colonies were individually toothpick-inoculated onto leaflets of Nicotiana benthamiana. 12 cDNAs were identified whose expression induced formation of a necrotic lesion around the inoculation site. 7 of these clones have different sequences. One of these clones PBC43 encodes specific elicitin. Clone PBC163 encodes a protein highly homologous to Rab; PBC241 en-codes a prohibitin protein; PBN62 encodes a Heat Shock Protein 60 (HSP60). The other five cDNAs reveal no homology to known protein and are thus considered novel. These observations suggest that this functional screening method is a versatile strategy to identify cDNAs of pathogens that encode elicitors and other HR-inducing proteins.We have devised a high-throughput functional cloning method to isolate cDNAs from Phylophthora boehmeriae of which the products elicit a hypersensitive response （HR） in tobacco. The cDNAs were cloned into a binary potato virus X （PVX）-based expression vector and transformed into Agrobacterium tumefeciens （Mog101）. 4100 colonies were individually toothpick-inoculated onto leaflets of Nicotiana benthamiana. 12 cDNAs were identified whose expression induced formation of a necrotic lesion around the inoculation site. 7 of these clones have different sequences. One of these clones PBC43 encodes specific elicitin. Clone PBC163 encodes a protein highly homologous to Rab; PBC241 encodes a prohibitin protein; PBN62 encodes a Heat Shock Protein 60 （HSP60）. The other five cDNAs reveal no homology to known protein and are thus considered novel. These observations suggest that this functional screening method is a versatile strategy to identify cDNAs of pathogens that encode elicitors and other HR-inducing proteins.

关 键 词：基因工程 克隆技术 敏感机制 烟草 基因编码 

分 类 号：S572[农业科学—烟草工业] Q785[农业科学—作物学]