叶锈菌诱导的小麦叶片cDNA文库的构建及其质量评价

Construction and Quality Identification of cDNA Library from Wheat Leaves Challenged by Puccinia recondita

由小麦叶锈菌(Puccinia triticina)引起的小麦叶锈病在世界各地产麦区均有发生,利用抗病品种是防治该病害最经济、安全、有效的方法。小麦抗叶锈基因Lr19是一个十分有效的抗叶锈基因,自1966年首次将该基因从长穗偃麦草转到普通小麦中,至今仍是一个应用潜力很大的抗病基因。本研究以小麦抗叶锈病近等基因系TcLr19和叶锈菌04-15-8①(生理小种类型THTS)为材料,构建叶锈病菌诱导的小麦叶片cDNA文库。研究结果表明文库的克隆数为4.1×106,插入片段大小在0.5～3.0kb,平均插入片段大小1.0kb。因此该文库可用于基因克隆与分析,为研究小麦抗叶锈病相关基因提供条件。

Wheat leaf rust caused by Puccinia triticina is a serious and widely distributed disease of wheat. Resistant cultivars are the most economic, safety and environmental friendly way for minimizing the losses caused by the disease. As an important leaf rust resistance gene, Lr19 was firstly transferred into the wheat genome from Thinopyrum sp. in the form of the substitution line ＂Agrus＂ in 1966. At present, Lr19 was a resistance gene with great potential to be used for wheat production. A cDNA library was constructed with wheat leaves challenged by Puccinia recondita is 04-15-8（1） by using cDNA library construction kit. The results showed that the number of library clones was 4.1 × 10^6. The clones of the library had an average insert size of 1.0 kb, which ranged 0.5-3.0 kb. Therefore this cDNA library is qualified for wheat gene cloning and function analysis.