杨树黑斑病感抗无性系cDNA文库的构建

Construction of cDNA Libraries with Leaves of Clones Susceptible and Resistant to Black Spot Disease in Poplar

为了分离与抗病相关的目的基因,以病原菌接种后对黑斑病具免疫力的美洲黑杨I 69及黑斑病的高感无性系欧美杨I 45的叶片为材料,在利用荧光差异显示反转录聚合酶链式反应(DDRT PCR)获得抗病相关cDNA片段的基础上,用试剂盒及同样的植物材料构建了两个cDNA文库L45 72及L69 72。两文库的滴度分别为3.44×109pfu/mL及3.94×109pfu/mL,大于500bp的片段分别占96.67%和73.33%。

In order to isolate target genes associated with disease resistance,two cDNA libraries viz.L45-72 and L69-72 were constructed on the basis of anti-disease cDNA fragments obtained by fluorescent DDRT-PCR with ready-to-use kits and pathogen-inoculated poplar leaves of I-69 and I-45,two clones either extremely resistant or susceptible to black spot disease in poplar.The library of I-45(L45-72) had a titer of 3.44×10~9 pfu/mL and the library of I-69(L69-72) had a titer of 3.94×10~9 pfu/mL,with inserts larger than 500 bp accounting for (96.67%) and 73.33%,respectively.The libraries constructed have laid a foundation for studies on the mechanism of disease resistance and disease susceptibility.