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.
Abstract
[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,LmERF17,LmERF18 and LmERF19.[Conclusion]Eight LmERFs(LmERF1,LmERF2,LmERF6,LmERF9,LmERF10,LmERF15,LmERF16 and LmERF19)were responded to organ development,cold stress,and ethylene-induced senescence,which provide the data for the subsequent excavation of target LmERFs for efficiently regulating secondary metabolism in L.macranthoides.
基金项目
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)
NETL
NSTL
万方数据