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基于mRNA差异显示技术的红球姜防御转录组分析

Defence transcriptome profiling of Zingiber zerumbet (L.) Smith by mRNA differential display.

作者信息

Kavitha P G, Thomas George

机构信息

Plant Molecular Biology Group, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram 695 014, India.

出版信息

J Biosci. 2008 Mar;33(1):81-90. doi: 10.1007/s12038-008-0002-2.

Abstract

Soft rot is a serious disease in ginger (Zingiber of?cinale Roscoe),imposing a considerable economic loss annually in all ginger-producing countries. In this study,mRNA differential display was employed to identify genes whose expression was altered in a soft rot-resistant accession of Zingiber zerumbet (L.) Smith,a wild relative of ginger, in response to Pythium aphanidermatum (Edson) Fitzp.,which is the principal causative agent of soft-rot disease in ginger. Analysis using 68 primer combinations identified 70 differentially expressed transcript-derived fragments (TDFs),of which 34 TDFs were selected for further analysis following reverse northern screening. Cloning and sequence characterization of the 34 TDFs yielded a total of 54 distinct clones. Functional categorization of these clones revealed seven categories,of which the defence/stress/signalling group was the largest,with clones homologous to genes known to be actively involved in various pathogenesis-related functions in other plant species.The significance of these genes in relation to the resistance response in Z.zerumbet is discussed. This study has provided a pool of candidate genes for detailed molecular dissection of the defence mechanisms in Z.zerumbet and for accessing wild genetic resources for the transgenic improvement of ginger.

摘要

软腐病是生姜(姜科姜属植物姜Zingiber officinale Roscoe)的一种严重病害,在所有生姜生产国都造成了相当大的经济损失。在本研究中,利用mRNA差异显示技术来鉴定在姜的野生近缘种红球姜(Zingiber zerumbet (L.) Smith)的一个抗软腐病材料中,响应生姜软腐病的主要病原菌瓜果腐霉(Pythium aphanidermatum (Edson) Fitzp.)而表达发生改变的基因。使用68对引物组合进行分析,鉴定出70个差异表达的转录本衍生片段(TDFs),其中34个TDFs在反向Northern杂交筛选后被选择用于进一步分析。对这34个TDFs进行克隆和序列特征分析,共获得54个不同克隆。对这些克隆进行功能分类,揭示出七个类别,其中防御/应激/信号传导组最大,有与已知在其他植物物种中积极参与各种病程相关功能的基因同源的克隆。讨论了这些基因与红球姜抗性反应的关系。本研究为深入分子剖析红球姜的防御机制以及为转基因改良生姜获取野生遗传资源提供了一批候选基因。

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