摘要
为探索在干旱后复水条件下糜子抗旱种质的基因表达特点,寻找与其复水后快速恢复生长相关的基因,利用抑制消减杂交(Suppressive subtraction hybridization,SSH)技术,以糜子干旱后复水处理叶片的cDNA为tester,干旱胁迫处理叶片的cDNA为driver,构建了一个抑制消减杂交文库。在文库中随机选取100个克隆(插入片段≥250 bp)测序,所获序列去除载体、引物及接头序列后,利用NCBI的BLAST序列比对分析软件分别对GenBank的dbEST数据库和非冗余蛋白数据库进行序列比对分析。序列同源性分析显示,文库中序列的同源基因主要以与新陈代谢、能量代谢、转录、蛋白加工、细胞信号转导及细胞组分生物合成相关的基因为主,其中以新陈代谢及细胞组分生物合成相关的基因数最多,分别占到与已知蛋白同源的EST的22.4%和10.3%。表明植物在干旱后复水条件下产生一系列的信号传导,同时启动相应的转录机制,激活响应基因的表达,以适应逆境的改变。
In order to investigate the gene expression profile of broomcorn millet （Panicum miliaceum L. ） under the condition of rehydration after serious drought and find out the key genes related to the recovering growth,a subtracted cDNA library of broomcorn millet was constructed by means of Suppressive Subtraction Hybridization （SSH）. The cDNA of broomcorn millet seedling leaves during rehydration after serious drought was used as the tester and cDNA from serious drought was used as the driver. One hundred clones with insertion equal or over 250 bp were randomly picked and sequenced. Then the sequences of vector,primers and adapters were removed from the EST sequences. The EST sequences were subjected to conduct similarity searches with Blast of NCBI to dbEST and non-redundance protein database in NCBI. The Blast result showed that the genes expressed during rehydration of serious drought in broomcorn millet mainly involved genes related to metabolism ,energy transformation, transcription, protein processing, cellular communication and signal transduction, as as well biogenesis of cellular components, the most genes were those related to metabolism and biogenesis of cellular components,holding 22.4% and 10. 3% in the known EST of protein homology respectively. Those genes were supposed to play important roles during the rehydration progress after serious drought and should be further tested. The research will facilitate the isolation and functional analysis of the key genes,so as to be used in the improvement of drought tolerance of other cereal crops.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2007年第3期81-86,共6页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家"863"计划项目(2006AA100201)
西北农林科技大学科研专项(20062R013)
关键词
抑制消减杂交(SSH)
糜子
复水
基因表达谱
SSH （suppressive subtraction hybridization）
broomcorn millet （Panicurn miliaceum L. ）
rehydration
gene expression profile
