中国农业科学 ›› 2008, Vol. 41 ›› Issue (5): 1267-1273 .doi: 10.3864/j.issn.0578-1752.2008.05.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦与条锈菌亲和互作的差减文库构建及初步分析

喻修道,屈志鹏,郭 军,于秀梅,黄雪玲,韩青梅,黄丽丽,康振生   

  1. 西北农林科技大学植物保护学院
  • 收稿日期:2007-03-11 修回日期:1900-01-01 出版日期:2008-05-10 发布日期:2008-05-10
  • 通讯作者: 康振生

Screening Differential Profile of Genes Expressed during Compatible Interactions between Puccinia striiformis and Wheat

  

  1. 西北农林科技大学植物保护学院
  • Received:2007-03-11 Revised:1900-01-01 Online:2008-05-10 Published:2008-05-10

摘要: 【目的】构建小麦与条锈菌亲和互作的差减文库,分离条锈菌侵染小麦过程中的差异表达基因。【方法】以小麦品种水源11和条锈菌相应毒性小种CY31号为材料,构建条锈菌侵染阶段的SSH-cDNA文库,挑选250个阳性克隆测序并进行生物信息学分析。【结果】聚类后得到149条非冗余EST(unigene)。经Blast X比对和功能分类分析,其中50条unigene(33.6%)未找到同源性匹配,25条(16.8%)与未知功能蛋白同源性较高;其余74条功能已知的unigene中,与初级代谢、能量相关的基因分别有13条和10个,占8.7%和6.7%,感病及防御相关的基因有6个约占4.0%;此外,获得两个与病原菌有较高同源性的基因。随后,进一步利用RT-PCR对6个基因的表达模式进行了分析。【结论】成功构建了小麦与条锈菌亲和互作的差减文库,分离出一部分与小麦条锈病发病相关的基因,可用于进一步研究条锈菌侵染过程中特异基因的功能。

关键词: 小麦, 条锈菌, 抑制差减杂交, 亲和互作, 反转录PCR

Abstract: 【Objective】Isolation of differentially expressed genes induced by Puccinia striiformis is the first step to reveal the molecular mechanism of interaction between wheat and yellow rust fungus. 【Method】A SSH-cDNA library was constructed with wheat variety shuiyuan-11 and a Chinese virulent race CY31 of Puccinia striiformis, 250 positive clones were selected for sequencing.【Result】In total 149 nonredundant ESTs were assembled with CAP3, then function annotation and classification were performed. There were 50 unigenes (33.6%) without or with weak similarities to the existent proteins in the nr database from NCBI and 25 unigenes (16.8%) significantly matched with hypothetical protein. Whilst the unigenes assigned to primary metabolism, energy and disease/defence accounted for 8.7%, 6.7%, 4.0%, respectively. Only 2 unigenes, however, had higher similarity with pathogenic protein. Four genes were randomly selected to investigate their expression profiles by reverse transcription PCR (RT-PCR). 【Conclusion】Some genes which had higher similarity with catalase, elongation factor and cell death suppressor protein were supposed to have important roles during compatible interaction between Puccinia striiformis and wheat.