明尼2761抗秆锈病相关基因的cDNA-AFLP分析

doi:10.3864/j.issn.0578-1752.2013.23.024

摘要/Abstract

摘要： 【目的】分析中国重要辅助鉴别寄主明尼2761经小麦秆锈菌诱导后的基因表达谱，寻找与其抗病基因表达相关的片段。【方法】运用cDNA-AFLP技术研究明尼2761和感病对照Thatcher分别接种小麦秆锈菌后的基因表达差异，对与明尼2761抗病相关的特异表达片段进行分析。【结果】140对AFLP引物组合能够检测到约7 000多条差异表达转录衍生片段（TDF），平均每一对引物可以获得50条条带，其中有86对引物能够在明尼2761和Thatcher之间扩增出差异条带。对可能与抗病相关的35条特异TDF进行克隆测序，经BLASTx比对，30条TDF在数据库中能够找到同源序列,其中24条TDF功能已知，6条TDF功能尚未明确。【结论】8条与编码激酶结合蛋白、NBS-LRR抗病蛋白、类肉桂酰-CoA还原酶2b、类肉桂酰辅酶A还原酶2c、蔗糖磷酸合成酶7、类蛋白酶metacaspase 1、类锌指蛋白、假定的类RGA4抗病蛋白基因有较高同源性的TDF应与明尼2761的抗秆锈性相关。

关键词: 小麦秆锈菌 , 明尼2761 , cDNA-AFLP , 抗病基因

Abstract: 【Objective】The objective of this study is to analyze the differential genes expression of the important Chinese supplementary differential Minn2761 induced by wheat stem rust, and to find out the fragments related to disease-resistant gene expression. 【Method】 The study was carried out by cDNA-AFLP analysis of differentially expressed resistant genes in Minn2761 and Thatcher inoculated with wheat stem rust.【Result】About 7 000 bands were amplified by 140 primer combinations and by average 50 bands appeared per pair of primers. Through cDNA-AFLP analysis, 86 primer combinations amplified specific bands between Minn2761 and Thatcher. Thirty-five possibly disease resistance-related transcript derived fragments (TDFs) were cloned and sequenced. BLASTx analysis showed that the sequences of 30 TDFs were found to be homologous with those in GenBank. Among them, 24 TDFs had known function and 6 TDFs had unknown function.【Conclusion】Eight TDFs having a highly homology with known genes which encoded kinase binding protein, NBS-LRR resistance protein, cinnamoyl CoA reductase-like 2b, cinnamoyl CoA reductase-like 2c, sucrose-phosphate synthase 7, metacaspase-1-like isoform 1, Zn-finger-like protein, and putative disease resistance protein RGA4-like were supposed to the resistance to stem rust of Minn2761.

Key words: wheat stem rust , Minn2761 , cDNA-AFLP , resistance gene

栾兆杰, 曹远银, 李天亚, 陈思, 陈秀梅, 朱桂清, 李伟华. 明尼2761抗秆锈病相关基因的cDNA-AFLP分析[J]. 中国农业科学, 2013, 46(23): 5058-5065.

LUAN Zhao-Jie, CAO Yuan-Yin, LI Tian-Ya, CHEN Si, CHEN Xiu-Mei, ZHU Gui-Qing, LI Wei-Hua. cDNA-AFLP Analysis of Differentially Expressed Resistant Genes of Minn2761[J]. Scientia Agricultura Sinica, 2013, 46(23): 5058-5065.

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