日本结缕草ZjPPH2基因的克隆、亚细胞定位及表达分析  

Cloning,Subcellular Localization Analysis and Expression Characterization of ZjPPH2 Gene from Zoysia japonica

作　　者：滕珂 李俊[2] 柳吉芹 岳跃森 范希峰[1] 武菊英[1] TENG Ke;LI Jun;LIU Ji-qin;YUE Yue-sen;FAN Xi-feng;WU Ju-ying(Beijing Research and Development Center for Grass and Environment,Beijing 100097,China;Institute of Grassland Research,Chinese Academy of Agricultural Sciences,Hohhot 010010,China;Qinhuangdao Entry-Exit Inspection and Quarantine Bureau,Qinhuangdao 066004,China)

机构地区：[1]北京草业与环境研究发展中心,北京100097 [2]中国农业科学院草原研究所,内蒙古呼和浩特010010 [3]秦皇岛出入境检验检疫局,河北秦皇岛066004

出　　处：《中国草地学报》2018年第4期1-7,共7页Chinese Journal of Grassland

基　　金：中国博士后基金(2017M620677);北京市博士后基金(2017-ZZ-087);北京市农林科学院博士后基金;北京市农林科学院科技创新能力建设专项(KJCX20161502-1)

摘　　要：利用RACE技术从日本结缕草中克隆得到1个脱镁叶绿素脱镁叶绿酸水解酶基因ZjPPH2(GenBank登录号:MG870086)。其开放阅读框为1140bp,编码379个氨基酸。构建系统进化树发现,ZjPPH2蛋白与高粱亲缘关系最近。通过染色体步移的方法得到ZjPPH2基因ATG上游1699bp序列,PLACE在线预测结果表明,其启动子序列上具有响应ABA、MeJA、SA的调控元件及若干光响应元件。亚细胞定位结果显示,ZjPPH2定位于叶绿体。荧光定量数据表明,ZjPPH2基因在衰老叶片中的表达量最高,且受ABA、MeJA、SA及黑暗处理的调控。The pheophytin pheophorbide hydrolase gene,ZjPPH2（GenBank No.MG870086）,was isolated from Zoysia japonica using RACE method.The open reading frame of ZjPPH2 gene was 1140 bp,encoding 379 amino acids.Phylogenetic analysis demonstrated that the ZjPPH2 was most closely related to that in Sorghum bicolor.A 1699 bp sequence from ATG of the ZjPPH2 gene was cloned through Genome Walking method.PLACE analysis indicated that the promoter sequence contained several cis-elements involved in MeJA,SA,ABA responsiveness and several photo-responsive components.Subcellular localization results showed that the ZjPPH2 was localized in chloroplasts.Real-time quantitative PCR analysis proved that ZjPPH2 gene was expressed most abundantly in senescent leaves.Moreover,the expression of ZjPPH2 could be regulated by exogenous MeJA,SA,ABA or dark treatment.This study proved that ZjPPH2 could be a functional gene in controlling chlorophyll degradation and paves the way to further study the ZjPPH2 in Zoysia japonica.

关 键 词：日本结缕草 PPH2基因 启动子 亚细胞定位 表达分析 

分 类 号：S688.4[农业科学—观赏园艺]