富集小麦磷胁迫响应基因的cDNA差减文库构建和部分ESTs的功能鉴定  

Construction of a cDNA Subtractive Library Enriched the Response Genes of Deficient-Pi Stress and Functional Identification of Some ESTs in the Library

作　　者：龙素霞[1] 路文静[2] 谷俊涛[2] 郭程瑾[1] 肖凯[1] 

机构地区：[1]河北农业大学农学院,河北保定071001 [2]河北农业大学生命科学学院,河北保定071001

出　　处：《华北农学报》2009年第2期12-16,共5页Acta Agriculturae Boreali-Sinica

基　　金：国家"973"项目(2007CB116209);河北省自然科学基金项目(C2007000476)

摘　　要：植物在形态学和生化代谢水平上对低磷胁迫逆境的响应，是特异表达基因在时空上精细协同作用的结果。以前期工作中鉴定的磷高效小麦品种石新828为材料，构建了富集不同低磷处理时间点特异表达基因的根系cDNA差减抑制杂交文库。获得的克隆总数为2682个。对随机选取的文库克隆研究发现，克隆中插入的片段长度为250．750bp。测序结果和功能比对发现，具有功能比对结果的克隆比例为70％，其中部分分别与小麦、大麦、水稻、玉米和拟南芥等植物种属高度同源，参与转录调控、蛋白质合成和代谢等多个生物学过程。该富集低磷胁迫响应基因文库为进一步鉴定小麦响应磷胁迫逆境的基因调控网络和克隆重要的磷高效相关基因奠定了坚实的基础。The responses of plant to deficient-Pi stress on phenotype and biochemical levels are the distinctly cooperative results resulting from the specific expressed genes at the spatiotemporal range. In this study, using Shixin828, the wheat eultivar with high phosphorus utilization efficiency identified previously to be the experimental material, a root cDNA selected-subtractive suppression library enriched the response genes of deficient-Pi stress at various time-points was constructed. The clone number in total was 2 682 in this library. It was found that the insertion lengths changed from 350 bp to 750 bp, by analyzing the randomly selected clones in the library. The clones with putative functions and the clones with unknown functions were comprised of the percentage of 70 % and 30 %, respectively. The former, being homogenous to wheat, barley, rice, maize, and Arabidopsis, owned the functions in the plant by involving in the transcriptional regula- tion, protein synthesis, and biochemical metabolism. Therefore, the root cDNA selected-subtractive suppression library will be useful for further identification of the gene-regulatory network to respond to the deficient-Pi and cloning of important genes related to forming the high-phosphorus utilization efficiency.

关 键 词：小麦 低磷胁迫 特异表达基因 cDNA差减抑制杂交文库 

分 类 号：S512.03[农业科学—作物学]