栽培小麦Brock白粉菌诱导早期应答基因SSH文库构建及分析

doi:10.7525/j.issn.1673-5102.2015.01.011

摘要/Abstract

摘要： 以接种白粉菌的Brock叶片cDNA作为实验组（tester），不接菌Brock叶片cDNA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GenBank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。

关键词: 栽培小麦Brock, 白粉病, 抑制差减杂交, 表达序列标签

Abstract: 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 cDNA 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, Lr1 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.

Key words: cultivar wheat Brock, powdery mildew, suppression subtractive hybridization(SSH), expression sequence tag(EST)

中图分类号:

乔石瑶；刘晓颖；王艳红；肖莹；孙世南；王振英*. 栽培小麦Brock白粉菌诱导早期应答基因SSH文库构建及分析[J]. 植物研究, 2015, 35(1): 60-67.

QIAO Shi-Yao；LIU Xiao-Ying；WANG Yan-Hong；XIAO Ying；SUN Shi-Nan；WANG Zhen-Ying*. Constructing and Analyzing SSH Library for the Early Response Genes in Wheat Brock after Blumeria graminis f. sp. tritici Stress[J]. Bulletin of Botanical Research, 2015, 35(1): 60-67.

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