蝴蝶兰亲环素基因PhCyP的克隆及表达分析  

Cloning and Expression of Cyclophilin Gene PhCyP from Phalaenopsis

作　　者：袁秀云[1] 许申平[1] 张燕[1] 王默霏[1] 蒋素华[1] 梁芳[1] 崔波[1] YUAN Xiuyun;XU Shenping;ZHANG Yan;WANG Mofei;JIANG Suhua;LIANG Fang;CUI Bo(Institute of Bioengineering,Zhengzhou Normal University,Zhengzhou,Henan 450044,China)

机构地区：[1]郑州师范学院生物工程研究所,河南郑州450044

出　　处：《热带作物学报》2020年第2期315-322,共8页Chinese Journal of Tropical Crops

基　　金：郑州师范学院科技创新团队支持计划项目(No.ZS180110);河南省高等学校重点科研项目(No.18A210005)。

摘　　要：为研究蝴蝶兰对低温胁迫分子调控的机理,采用RT-PCR和RACE方法,从蝴蝶兰叶片中克隆到一个亲环素基因PhCyP(登录号为MH992514),其基因cDNA全长序列为829 bp,开放阅读框长度为522 bp,编码173个氨基酸,其编码蛋白含有一个类亲环素(CLD)保守结构域,为碱性亲水性蛋白。系统进化分析显示,蝴蝶兰PhCyP蛋白与兰科植物小兰屿蝴蝶兰、万带兰、深圳拟兰、铁皮石斛等亲环素蛋白亲缘关系较近。转录表达分析表明,PhCyP基因在蝴蝶兰不同发育时期的根、叶、花梗、花器官、子房、种子等组织中有高丰度表达。在13℃/8℃(昼/夜)温度条件下,PhCyP基因的表达水平先降低,处理6 d时降至最低,随后逐渐升高至恢复培养。在4℃低温条件下,PhCyP基因的表达水平升高,在处理4 h升至最高,然后逐渐下降,在处理48 h其表达低于处理前水平。以上结果表明,PhCyP基因参与蝴蝶兰对低温胁迫的响应,且对不同低温胁迫具有不同的分子调控机制。To study the molecular regulation mechanism of Phalaenopsis in response to low temperature stress,a cyclophilin gene PhCyP(GenBank accession number:MH992514)was cloned from the leaf of Phalaenopsis through the RT-PCR and RACE method.The full-length cDNA sequence was 829 bp with an open reading frame(ORF)of 522 bp encoding 173 amino acids.PhCyP contained a cyclophilin-like domain(CLD)with alkaline and hydrophilic properties.The phylogenetic tree showed that PhCyP was closely related to the cyclophilin from Phalaenopsis equestris,Vanda hybrid cultivar,Apostasia shenzhenica and Dendrobium catenatum in Orchidaceae.The expression analysis showed that PhCyP was highly expressed in different tissues including root,leaf,flower,ovary and seed in different development stage.Under 13℃/8℃ day/night temperature,the expression level of PhCyP reduced gradually with treatment time and was the lowest at 6 d,then increased gradually from 9 d to restoring.Under 4℃treatment,the expression level of PhCyP increased gradually,and achieved the highest level at 4 h,then its expression level decreased gradually with a lower level than that before treatment at 48 h.The results suggested that PhCyP was involved in the response to low temperature stress with different molecular regulation mechanism to different low temperature stress.

关 键 词：蝴蝶兰 低温 亲环素 基因表达 

分 类 号：S682.31[农业科学—观赏园艺] Q786[农业科学—园艺学]