春兰组培苗镁原卟啉IX甲基转移酶基因克隆及功能分析  

Molecular Cloning and Functional Analysis of Magnesium ProtoporphyrinⅨMethyltransferase Gene in Tissue Culture Seedlings of Cymbidium goeringii

作　　者：蒋彧[1] 涂勋良 何俊蓉[1] JIANG Yu;TU Xunliang;HE Junrong(Institute of Horticulture,Sichuan Academy of Agricultural Sciences,Chengdu 610000,China)

机构地区：[1]四川省农业科学院园艺研究所,四川成都610000

出　　处：《四川林业科技》2022年第4期83-89,共7页Journal of Sichuan Forestry Science and Technology

基　　金：四川省重点研发项目(2021YFN0118);省财政自主创新专项(2022ZZCX047);成都市技术创新研发项目(2022-YF05-00707-SN);现代农业产业技术体系专项资金资助(CARS-23-G58);四川省育种攻关(2021YFYZ0006);科技成果中试熟化(2021ZSSFKJ51)。

摘　　要：镁原卟啉IX甲基转移酶(ChlM)是叶绿素合成途径中的关键限速酶之一,在植株生长发育过程中发挥着重要作用。为了了解春兰中ChlM的功能,从春兰‘宋梅’组培苗中克隆了ClChlM1基因。该基因开放阅读框全长945 bp,编码314个氨基酸序列。序列比对显示ClChlM1含有Mg-por_mtran_C结构域,亚细胞定位结果显示该基因定位在叶绿体上。在烟草中过量表达ClChlM1,叶片叶绿素含量及ALA比野生型增加。另外,过量表达烟草叶片面积也大于野生型。在过量表达转基因株系中,编码谷氨酸酯-1-半醛2,1-氨基变位酶(Gsa),叶绿素a/b结合蛋白(LHCB),镁离子螯合酶CHLI和镁离子螯合酶CHLH的基因表达上调。The magnesium protoporphyrin IX methyltransferase(ChlM)is one of the key rate-limiting enzymes in chlorophyll synthesis and development of chloroplasts,which plays an important role in plant growth and development.To better understand the role of ChlM in Cymbidium goeringii,ChlM gene was cloned from the tissue culture seedling of Cymbidium goeringii‘Song Mei’.The full-length open reading frame of ClChlM1 was 945 bp and encoded 314amino acids.The sequence alignment showed that ClChlM1 contained an Mg-por_mtran_C domain,and subcellular location analysis indicated ClChlM1 was located in chloroplast.Furthermore,the overexpression of ClChlM1 in tobacco leaves could significantly increase the content of chlorophyll and ALA compared with wild type tobacco leaves.In addition,the leaf area of overexpressed tobacco was larger than that of wild type.In the over-expressed transgenic lines,the gene expression of glutamate-1-semialdehyde 2,1-aminomutase(Gsa),chlorophyll a/b binding protein(LHCB),magnesium chelatase CHLI and magnesium chelatase CHLH were up-regulated.

关 键 词：兰属春兰 组培 叶绿素 镁原卟啉IX甲基转移酶基因 

分 类 号：S682.31[农业科学—观赏园艺]