棉花亲环素基因GhCYP2的克隆及实时定量表达分析  

Cloning of Cyclophilin(GhCyp2)in Gossypium hirsutum and Its Quantitative Analysis by Real-Time RT-PCR

作　　者：吴莉[1] 王雅琴[1] 张新宇[2] 孙杰[2] 李艳军[2] 

机构地区：[1]石河子大学生命科学学院,新疆石河子832003 [2]石河子大学农学院,新疆石河子832003

出　　处：《新疆农业科学》2011年第9期1569-1575,共7页Xinjiang Agricultural Sciences

基　　金：农业部转基因生物新品种培育重大专项(2008Zx08005)

摘　　要：【目的】分离棉花中的亲环素基因。亲环素基因是一类多基因蛋白质家族,广泛存在于高等植物各个组织和器官中,参与多种重要的生理生化过程。【方法】以小麦亲环素wCyP3基因蛋白序列(AF262984)为探针序列,在GenBank中与棉花EST序列进行tBLASTn比对,将同源性高的EST序列拼接后获得一条长912 bp的cDNA序列。【结果】序列分析表明,该cDNA含有一个编码174个氨基酸的开放阅读框,基因的编码产物为亲环素蛋白,将该基因命名为GhCYP2(GenBank登录号:JN032296)。采用Clustal X将GhCYP2的氨基酸序列与其他生物亲环素氨基酸序列进行相似性比对,发现与水稻、拟南芥等生物的相似性较高,达到80%以上。利用实时荧光定量PCR技术分析了GhCYP2在棉花各个组织器官中的表达模式,检测结果显示:GhCYP2在棉花叶及18DPA棉纤维中的表达量最高,在根、茎、花、3～15DPA、21～30DPA棉纤维中均有适量表达。【结论】推测GhCYP2可能在棉花纤维伸长至次生壁合成的重叠期发挥着信号传导作用。[ Objective] Cyclophilins（CyPs） are supper - family members and ubiquitously distributed in diverse organs and tissues of higher plant, and involved in numerous physiological and biochemical processes. [ Method ] Possible CYP sequences in Gossypium hirsutum were identified among plant expressed sequence tag （EST） by querying the EST database with the wCyP3 protein sequence using the tBLASTn procedure. ESTs similar to wCyP3 were identified, aligned and assembled into a 912 bp cDNA sequence. The sequence analysis indicated the cDNA contained a putative ORF encoding a cyclophilin protein of 174 amino acids, and the cDNA was named as GhCYP2 （GenBank accession：JN032296）. [ Result ] CLUSTAL X procedure analysis showed that the similarity was greater than 80% between cyclophilin genes of cotton and Arabidopsis, Oryza etc. The expression pattern was analyzed in cotton to determine its physiological function. QRT - PCR analysis showed that GhCYP2 had high expression in leaves and 18DPA fiber, and had moderate expression in root, stem, flower, and 3 -15DPA, 21 -30 DPA cotton fiber. [ Conclusion]The results presumed that GhCYP2 may play an important role for signal transduction in the period of overlap between fiber elongation and secondary wall synthesis.

关 键 词：棉花 亲环素 实时荧光定量PCR 

分 类 号：S562[农业科学—作物学] S188.2