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

doi:10.7668/hbnxb.2009.02.003

华北农学报 ›› 2009, Vol. 24 ›› Issue (2): 12-16. doi: 10.7668/hbnxb.2009.02.003

所属专题： 小麦；生物技术；

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

龙素霞¹, 路文静², 谷俊涛², 郭程瑾¹, 肖凯¹

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

收稿日期:2008-10-10 出版日期:2009-04-28
通讯作者: 肖凯( 1963- ), 男, 河北秦皇岛人, 教授, 博士生导师, 主要从事作物栽培和生理的研究。
作者简介:龙素霞( 1984- ), 女, 河北邯郸人, 在读硕士, 主要从事作物高产与分子生物学研究.
基金资助:
国家"973"项目(2007CB116209);河北省自然科学基金项目(C2007000476)

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

LONG Su-xia¹, LU Wen-jing², GU Jun-tao², GUO Cheng-jin¹, XIAO Kai¹

1. College of Agronomy, Agricultural University of Hebei, Baoding 071001, China;
2. College of Life Science, Agricultural University of Hebei, Baoding 071001, China

Received:2008-10-10 Published:2009-04-28

摘要/Abstract

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2 682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20～70 bp.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.

关键词: 小麦, 低磷胁迫, 特异表达基因, cDNA差减抑制杂交文库

Abstract: The responses of plant to deficient-Pistress 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 cultivar with high phosphorus utilization efficiency identified previously to be the experimental material,a root cDNA selected2subtractive suppression library enriched the response genes of deficient2Pi stress at various time-points was constructed.The clone number in total was 2682 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 regulation,protein synthesis,and biochemical metabolism.Therefore,the root cDNA selected-subtractive suppression library will be useful for further identification of the gene2regulatory network to respond to the deficient-Pi and cloning of important genes related to forming the high-phosphorus utilization efficiency.

Key words: Wheat(Triticum aestivum L. ), Deficient-Pi stress, Specific expressed gene, cDNA selected-subtractive suppression library

中图分类号:

S12.03

龙素霞, 路文静, 谷俊涛, 郭程瑾, 肖凯. 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[J]. 华北农学报, 2009, 24(2): 12-16. doi: 10.7668/hbnxb.2009.02.003.

LONG Su-xia, LU Wen-jing, GU Jun-tao, GUO Cheng-jin, XIAO Kai. Construction of a cDNA Subtractive Library Enriched the Response Genes of Deficient-Pi Stress and Functional Identification of Some ESTs in the Library[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(2): 12-16. doi: 10.7668/hbnxb.2009.02.003.

http://www.hbnxb.net/CN/Y2009/V24/I2/12

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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

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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

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