摘要
以晚疫病病原菌混合小种接种处理 4 8h的马铃薯水平抗性材料 (R gene free)叶片为目的材料 ,以未处理材料作为对照 ,用抑制差减杂交技术构建了一个富集晚疫病抗性相关基因的差减文库。应用反向Northern技术对84 0个克隆进行斑点杂交筛选 ,筛选出 15 0个病原诱导后信号明显增强的克隆。 2 6个片段测序结果表明 :部分片段基因功能与抗病性明显相关。 7个差异表达片段与GenBankEST数据库中已有晚疫病原诱导马铃薯叶片得到的EST有很高同源性 (达 95 %～ 10 0 % ) ;部分片段核苷酸或氨基酸序列分别与番茄、烟草、拟南芥等的EST序列或氨基酸序列有较高同源性 ;另有 4个基因片段在GenBankEST数据库中未找到明显的同源序列 ,可能为新发现的基因片段。
Suppression subtractive hybridization (SSH) was applied in the search of the genes induced during the interaction between Phytophthora infestans and horizontal resistance potato (R gene free).A subtractive library of potato,which highly enriches for resistance related genes to P.infestans ,was constructed.The potato leaves after inoculating 48 h with mixture of P.infestans races were used as 'tester' and uninoculated leaves as 'control' in the experiments.Total 150 of 840 randomly chosen cDNA clones were identified with the reveres Northern dot blot screening method.The sequencing results of 26 clones indicated that several fragments were previously shown to be activated during resistance responses.Seven differentially expressed fragments had significant homologous with EST of potato leaves inoculated with P.infestans .The sequences of nucleotide or amino acids of partial cDNA fragments shared identity with tomato,tobacco and Arabidopsis while four cDNA fragments that had no significant homology with EST in GenBank were expected to be new genes involved in the disease response.
基金
国家自然科学基金项目 (编号 :3 9970 464 )
国家"863"基金项目 (编号 :2 0 0 2AA2 41181)资助~~
