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真菌激发子和创伤处理的野生稻(大颖野生稻)差异表达转录本的分析

Analysis of differentially expressed transcripts of fungal elicitor- and wound-treated wild rice (Oryza grandiglumis).

作者信息

Kim Kyung Mi, Cho Sung Ki, Shin Sang Hyun, Kim Gyung-Tae, Lee Jai Heon, Oh Boung-Jun, Kang Kyung Ho, Hong Jong Chan, Choi Jun Young, Shin Jeong Sheop, Chung Young Soo

机构信息

Department of Genetic Engineering, Dong-A University, Pusan, South Korea.

出版信息

J Plant Res. 2005 Oct;118(5):347-54. doi: 10.1007/s10265-005-0228-0. Epub 2005 Sep 1.

Abstract

Suppression subtractive hybridization was used to construct a subtracted library (ogfw) from plants of a wild rice species, Oryza grandiglumis, subjected to a fungal elicitor and physical wounding. To screen the differentially expressed transcripts in the library, we applied a reverse Northern blot analysis to a cDNA microarray containing 1,152 random clones. Based on the average expression ratio, we selected 156 clones showing an elevated expression level. The elevated expression levels and overall expression profiles over time were verified by Northern blot analysis. A comparative functional categorization of the subtracted expressed sequence tags (ESTs) of the ogfw library against ESTs isolated from blast-infected O. sativa showed that the functional categories of cell rescue, defense and virulence, transcription, and cellular transport and transport mechanism of the ogfw library were threefold higher in the former than in the latter. These subtracted ESTs can be presumed to be related to the defense/resistance system and will be used to investigate the defense mechanisms of wild rice and to provide new insights into the genome of wild rice, which in turn will assist molecular breeding strategies of cultivated rice.

摘要

抑制性消减杂交技术被用于构建一个消减文库(ogfw),该文库来自受真菌激发子和物理创伤处理的野生稻物种大颖野生稻(Oryza grandiglumis)的植株。为了筛选文库中差异表达的转录本,我们对包含1152个随机克隆的cDNA微阵列进行了反向Northern印迹分析。基于平均表达率,我们选择了156个表达水平升高的克隆。通过Northern印迹分析验证了这些克隆表达水平的升高以及随时间的整体表达谱。将ogfw文库的消减表达序列标签(EST)与从稻瘟病感染的栽培稻中分离的EST进行比较功能分类,结果表明,ogfw文库在细胞拯救、防御和毒力、转录以及细胞运输和运输机制等功能类别上,前者比后者高出三倍。这些消减的EST可以推测与防御/抗性系统相关,并将用于研究野生稻的防御机制,为野生稻基因组提供新的见解,进而有助于栽培稻的分子育种策略。

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