贡菊CmNAC78的克隆及启动子元件分析  

Cloning and Promoter Element Analysis of CmNAC78 in Chrysanthemum'Gongju'

作　　者：岳圆圆 赵丽 全英杰 任馒蓉 杨雯婷 何子涵 田鑫 高日[1] Yue Yuanyuan;Zhao Li;Quan Yingjie;Ren Manrong;Yang Wenting;He Zihan;Tian Xin;Gao Ri(Agricultural College of Yanbian University,Yanji,133400;Yanbian Academy of Forestry Sciences,Yanji,133002)

机构地区：[1]延边大学农学院,延吉133400 [2]延边林业科学研究院,延吉133002

出　　处：《分子植物育种》2021年第14期4618-4625,共8页Molecular Plant Breeding

基　　金：吉2018年“十三五”科学技术项目(JJKH20180902KJ);延大科合字(2017)第21号;延边大学博士科研启动基金(2018)共同资助

摘　　要：NAC是植物中特有的一类转录因子,对植物生长发育和抵御非生物胁迫起着重要的作用。本试验以贡菊为材料,克隆CmNAC78基因,其最大开放阅读框(ORF)为1641 bp,编码547个氨基酸,包含A、B、C、D、E五个结构域;CmNAC78分子量为1354.64 kD,理论等电点为5.01,不稳定系数为38.37,平均亲水性0.649,脂肪系数为27.24,亚细胞定位预测CmNAC78定位在细胞核内。进化树分析表明,贡菊CmNAC78属于NAC2亚家族,与拟南芥ANAC78亲缘关系较近;采用染色体步移试剂盒方法,克隆了CmNAC78启动子序列2034 bp,启动子序列中包括茉莉酸甲酯、组织器官发育和低温响应元件等;荧光定量PCR分析不同花期CmNAC78、CmFNS和CmTTG1的相对表达量,结果表明在花蕾期时CmNAC78、CmFNS和CmTTG1表达量最高,其次为初花期。这些研究结果为后续在菊花中CmNAC78的调控机制研究提供参考。NAC is a unique type of transcription factor in plants,which plays an important role in plant growth and development and resistance to abiotic stress.In this experiment,the CmNAC78 gene was cloned from Chrysanthemum'Gongju'.Its maximum open reading frame(ORF)is 1641 bp,encoding 547 amino acids,and it contains five domains:A,B,C,D,and E.The molecular weight of CmNAC78 is 1354.64 kD,heoretical pI is 5.01,the instability index is 38.37,the average hydrophilicity is 0.649,the aliphatic index is 27.24.The subcellular localization predicts that CmNAC78 is localized in the nucleus.Phylogenetic tree analysis showed that CmNAC78 belongs to the NAC2 subfamily and has a close relationship with ANAC78 of Arabidopsis thaliana.CmNAC78 promoter sequence was cloned 2034 bp by chromosome step kit method.The promoter sequence included methyl jasmonate,Tissue and organ development and low temperature response elements;The relative expression levels of CmNAC78,CmFNS and CmTTG1 at different flowering stages were analyzed by real-time quantitative PCR.The results showed that the expression levels of CmNAC78,CmFNS and CmTTG1 were highest at the budding stage,followed by the initial flowering stage.These results provide reference for the subsequent research on the regulation mechanism of CmNAC78 in chrysanthemum.

关 键 词：贡菊 启动子 相对表达量 CmNAC78 

分 类 号：S682.11[农业科学—观赏园艺]