灰毡毛忍冬ERF基因家族鉴定及其在衰老过程中的表达分析  

Identification of the ERF gene family in Lonicera macranthoides and its expression profiles during senescence

作　　者：刘燕妮 曹正艳 吴佩荫 李清清 陈泽雄[2,3] 唐宁 LIU Yanni;CAO Zhengyan;WU Peiyin;LI Qingqing;CHEN Zexiong;TANG Ning(College of Biology and Food Engineering,Chongqing Three Georges University,Wanzhou 404100,Chongqing,China;College of Smart Agriculture,Chongqing University of Arts and Sciences,Yongchuan 402160,Chongqing,China;Institute of Special Plants,Chongqing University of Arts and Sciences,Yongchuan 402160,Chongqing,China)

机构地区：[1]重庆三峡学院生物与食品工程学院,重庆万州404100 [2]重庆文理学院智慧农业学院,重庆永川402160 [3]重庆文理学院特色植物研究院,重庆永川402160

出　　处：《经济林研究》2024年第2期13-28,共16页Non-wood Forest Research

基　　金：国家自然科学基金项目(32270401);重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX1196);重庆高校创新研究群体项目(CXQT21028)。

摘　　要：【目的】灰毡毛忍冬Lonicera macranthoides为我国南方地区大宗道地中药材和重要经济植物,绿原酸和木犀草苷等药效物质在器官衰老过程中积累且受转录因子调控。乙烯信号转导相关ERF基因家族广泛参与植物的衰老、胁迫响应和次生代谢。鉴定灰毡毛忍冬ERF家族成员并探究其表达模式,可为该类基因调控次生代谢的功能研究提供参考。【方法】基于转录组数据鉴定灰毡毛忍冬ERF转录因子家族,通过系统发育树分析其进化关系,利用MEME在线网站分析序列的结构基序。QRT-PCR分析LmERFs基因在不同器官衰老过程以及冷胁迫和乙烯处理后的表达模式。【结果】在灰毡毛忍冬中共鉴定出65个具AP2保守结构域的LmERFs基因,可将其分为3个亚组。利用RNA-Seq技术筛选了在幼嫩和衰老叶片之间差异表达的20个LmERFs。进一步qRT-PCR分析显示,分别有18个、13个和11个LmERFs基因在叶片、茎和花的衰老过程显著差异表达,其中LmERF2、LmERF9、LmERF16、LmERF17和LmERF19基因表达在不同器官衰老过程中均具显著差异。冷胁迫后,叶和茎中7个LmERFs(LmERF1、LmERF2、LmERF6、LmERF7、LmERF8、LmERF11和LmERF12)显著上调,LmERF3、LmERF9、LmERF16和LmERF19显著下调;而乙烯处理诱导了LmERF1、LmERF8和LmERF12基因的表达,抑制了LmERF2、LmERF4、LmERF9、LmERF10、LmERF14、LmERF15、LmERF16、LmERF17、LmERF18和LmERF19基因的表达。【结论】8个LmERFs(LmERF1、LmERF2、LmERF6、LmERF9、LmERF10、LmERF15、LmERF16和LmERF19)响应了器官发育、冷胁迫和乙烯诱导的衰老过程,这为后续挖掘高效调控灰毡毛忍冬次生代谢的靶点LmERFs提供了数据支持。【Objective】Lonicera macranthoides is a major authentic Chinese herb and an important economic plant in southern China.Its secondary metabolites,such as chlorogenic acid and luteoloside,accumulate during organ senescence and are regulated by transcription factors.Ethylene signaling related ERF gene family plays an important role in plant senescence,stress responses and secondary metabolism.Identification of ERF family members and exploration of their expression patterns can provide a reference for functional studies on the regulation of secondary metabolism by this class of genes.A comprehensive study of the ERF gene family in L.macranthoides has not yet been reported.【Method】The ERF transcription factor family was identified based on transcriptomic data.A phylogenetic tree was constructed to analyze their evolutionary relationships,and the structural motifs of these sequences were analyzed using the MEME Suite tools.QRT-PCR was used to analyze the expression patterns of LmERF genes in different organs during senescence as well as under cold stress and ethylene treatment.【Result】A total of 65 LmERF genes with conserved structural domains of AP2 were identified in L.macranthoides,which were divided into three subgroups.20 LmERFs were observed to be differentially expressed between young and senescent leaves using RNA-Seq.Further qRT-PCR analysis showed that 18,13 and 11 LmERF genes were significantly differentially expressed during senescence in leaves,stems,and flowers,respectively.Of them,LmERF2,LmERF9,LmERF16,LmERF17 and LmERF19 were remarkably differentially expressed during senescence in all tissues.After low temperature treatment,seven LmERFs,including LmERF1,LmERF2,LmERF6,LmERF7,LmERF8,LmERF11 and LmERF12,were strongly up-regulated,while LmERF3,LmERF9,LmERF16 and LmERF19 were outstandingly down-regulated in leaves and stems.Ethylene treatment induced the expression levels of LmERF1,LmERF8 and LmERF12,while suppressed the expression levels of LmERF2,LmERF4,LmERF9,LmERF10,LmERF14,LmERF15,LmERF16,Lm

关 键 词：灰毡毛忍冬 ERF基因家族 器官衰老 乙烯 冷害胁迫 

分 类 号：S567.79[农业科学—中草药栽培]