中国水仙NtTFL1-1和NtTFL1-2基因的克隆和表达  

Cloning and expression of the NtTFL1-1 and NtTFL1-2 genes from Narcissus tazetta var.chinensis

作　　者：张璐 吴菁华[1] 姚馨婷 王革伏 张志忠[1] ZHANG Lu;WU Jinghua;YAO Xinting;Gefu WANG-PRUSKI;ZHANG Zhizhong(Joint FAFU-Dalhousie Lab,College of Horticulture,Fujian Agriculture and Forestry University(FAFU),Fuzhou 350002,China;Faculty of Agriculture,Dalhousie University,Truro B2N 5E3,Canada)

机构地区：[1]福建农林大学园艺学院,福建农林大学-戴尔豪西联合实验室,福州350002 [2]加拿大戴尔豪西大学农学院,加拿大特鲁罗B2N5E3

出　　处：《应用与环境生物学报》2020年第2期264-271,共8页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(31601783);福建省级扶贫开发工作重点县人才支持计划专项(K1519060A);福建农林大学创新基金项目(CXZX2017533)资助。

摘　　要：中国水仙(Narcissus tazetta var. chinensis)是十大传统名花之一,一般在冬季开花,为典型的年宵花卉,开花时间的局限使其应用价值受到一定限制.水仙花球在夏季的休眠期进行花芽分化,是花期调控研究的重点阶段,但目前对其开花机制和调控的研究尚处于起步阶段. TFL1(TERMINAL FLOWER 1)是植物重要的开花相关基因,为了解该基因在中国水仙花芽分化和开花过程中的作用,以中国水仙‘金盏银台’为材料,利用逆转录聚合酶链式反应(RT-PCR,Reverse Transcription-Polymerase Chain Reaction)技术克隆中国水仙的TFL1基因NtTFL1-1和NtTFL1-2,对获得的基因序列进行相关生物信息学分析,对该基因编码蛋白进行亚细胞定位;利用荧光定量PCR技术对其在中国水仙花芽分化不同时期、不同器官类型中的差异表达情况进行测定.结果显示NtTFL1-1开放阅读框(open reading frame,ORF)共501个碱基对(bp,base pair),编码166个氨基酸,NtTFL1-2的ORF共525 bp,编码174个氨基酸;二者均具有TFL1家族的保守结构域;NtTFL1-1和NtTFL1-2均定位在细胞质中. NtTFL1-1和NtTFL1-2在中国水仙花芽分化初期和花序原基形成期的表达量较低,在花原基形成期表达量较高;NtTFL1-1和NtTFL1-2在根中的表达量高于茎、叶和成熟的花.本研究显示NtTFL1-1和NtTFL1-2和中国水仙花芽分化密切相关,结果可为中国水仙花期调控研究提供理论参考.(图6表2参30)Narcissus tazetta var.chinensis(Nt)is one of the ten most famous traditional flowers of China.It usually blooms in winter and is a typical flower of the Spring Festival.However,fixed florescence limits its applications.The flower bud differentiation of Nt occurs during the dormancy period of the bulb in summer.This period represents the key stage for flowering regulation research.Nonetheless,flowering mechanism and regulation research is still in its infancy.TFL1 is an important flowering-related gene in plants.In order to reveal the role of the TFL1 gene in flower bud differentiation and flowering in Nt,we cloned the TFL1 gene from Nt and carried out bioinformatics analysis.Both the TFL1 protein subcellular localization and TFL1 gene expression level were analyzed.The results will provide references for florescence regulation and the breeding of Nt varieties with different fluorescence.The TFL1 gene of Nt was cloned by reverse transcription-polymerase chain reaction(RT-PCR)using Nt cv.‘Jinzhanyintai’.Two clones were obtained in this experiment and named NtTFL1-1 and NtTFL1-2.The obtained gene sequence underwent bioinformatics analysis,and a subcellular localization of the gene encoding protein was carried out.The differential expression of the gene during the different stages of flower bud differentiation and the different organ types of Nt were determined by qPCR.Bioinformatics analysis showed that the open reading frame(ORF)of NtTFL1-1 was 501 bp,encoding 166 amino acids,and the ORF of NtTFL1-2 was 525 bp,encoding 174 amino acids,all of which had conserved domains of the TFL1 family.NtTFL1-1 and NtTFL1-2 were localized in the cytoplasm.The results of the qRT-PCR analysis showed that the expression of the NtTFL1-1 and NtTFL1-2 genes was lower during the early stages of flower bud differentiation and inflorescence primordium formation and higher during flower primordium formation.The expression of NtTFL1-1 and NtTFL1-2 in the roots was significantly higher than that in the stems,leaves,or mature flowers.The

关 键 词：中国水仙 TFL1基因 花芽分化 荧光定量PCR 亚细胞定位 

分 类 号：S682.21[农业科学—观赏园艺] Q943.2[农业科学—园艺学]