栽培小麦Brock白粉菌诱导早期应答基因SSH文库构建及分析

Constructing and Analyzing SSH Library for the Early Response Genes in Wheat Brock after Blumeria graminis f. sp. tritici Stress

以接种白粉菌的Brock叶片c DNA作为实验组(tester),不接菌Brock叶片c DNA为驱动组(driver),利用抑制性差减杂交技术,构建Brock白粉菌诱导早期应答基因消减杂交文库。随机挑选50个阳性克隆测序,获得38条非重复序列。经与Gen Bank进行同源比对,发现其中28条非重复序列与已知序列同源性较高。利用荧光定量PCR对部分基因分析后,发现ABC转运蛋白、丝/苏氨酸蛋白激酶、Lr1基因编码蛋白、核酮糖二磷酸羧基酶和叶绿素a/b结合蛋白参与了小麦抗白粉菌早期应答反应。该工作为我们在整体水平上揭示Brock抗白粉病分子机理提供重要依据。

We constructed a suppression subtractive hybridization （SSH） library for the early response genes in wheat Brock under Blumeria graminis f. sp. tritici （Bgt） stress by using the eDNA from Bgt-inoculated Brock leaf as the tester and the cDNA from Bgt-uninoculated Brock leaf as the driver. Fifty positive clones were picked randomly and sequenced, 38 redundant sequences were acquired. By sequence comparison against the GenBank database, 28 sequences was homolog of known genes. The transcriptional levels were assayed by fluorescence quantitative PCR, and the ABC transporter, serine/threonine protein kinase, Lrl gene-related protein, Ribulose bisphosphate carboxylase and Chlorophyll a/b binding protein were involved in the early response to Bgt stress in wheat cultivar Brock. Our work was useful for the powdery mildew resistance mechanism in wheat.