乙烯利诱导菠萝成花过程中FT基因的克隆与表达分析  

Expression and analysis of FT genes in pineapple flowering induced by ethephon

作　　者：阮城城 年宇薇 胡福初[2,3] 王祥和 范鸿雁[2,3] 吴凤芝 陈哲[2,3] 张治礼 RUAN Chengcheng;NIAN Yuwei;HU Fuchu;WANG Xianghe;FAN Hongyan;WU Fengzhi;CHEN Zhe;ZHANG Zhili(Institute of Tropical Crops,Hainan University,Haikou 570228,Hainan,China;Institute of Tropical Fruit Trees,Hainan Academy of Agricultural Sciences/Key Laboratory of Tropical Fruit Tree Biology of Hainan Province,Haikou 571100,Hainan,China;Investigation Station of Tropical Fruit Trees of Ministry of Agriculture,Haikou 571100,Hainan,China;Institute of Life Sciences and Pharmacy,Hainan University,Haikou 570228,Hainan,China)

机构地区：[1]海南大学热带作物学院,海口570228 [2]海南省农业科学院热带果树研究所·海南省热带果树生物学重点实验室,海口571100 [3]农业部海口热带果树科学观测实验站,海口571100 [4]海南大学生命科学与药学院,海口570228

出　　处：《果树学报》2019年第12期1648-1657,共10页Journal of Fruit Science

基　　金：海南省科协青年科技英才学术创新计划项目(QCXM201803);国家自然科学基金项目(31260460,31960589);海南省农业科学院农业科技创新专项

摘　　要：【目的】克隆菠萝AcFT基因并研究其表达模式,为深入研究该基因在乙烯利诱导菠萝成花中的功能奠定基础。【方法】从菠萝基因组数据库中获得AcFT基因全长序列,设计全长引物,克隆两个AcFT基因,分别命名为AcFT1和AcFT2,对基因序列进行生物信息学分析;通过qRT-PCR探究乙烯利处理后菠萝AcFT基因在不同组织、时间下的表达模式。【结果】AcFT1和AcFT2分别编码178和177个氨基酸,两者均含有PBP结构域,具有PEBP家族典型结构特征。实时荧光定量PCR分析结果显示,AcFT1和AcFT2都具有组织表达特异性,在茎和叶中相对表达量较高;乙烯利处理后,AcFT1和AcFT2在茎和叶中的表达具有相反的趋势,其中AcFT2明显受到乙烯利上调,呈现先上升后下降趋势,AcFT1则显示为先下降后上升。乙烯利处理后1 d,茎尖组织AcFT2的表达水平显著上升,相对表达量约为对照的178倍,而AcFT1则明显下调;乙烯利处理后31 d,AcFT2在茎尖中的表达水平达到最大值,约为对照的408倍,但AcFT1却处于极低水平。【结论】克隆得到2个AcFT基因,AcFT2基因在乙烯利处理后1 d及随后的花芽分化时期高度表达,表明AcFT2在响应外源乙烯利信号诱导菠萝成花过程中发挥重要作用。【Objective】The FT(Flowering Locus T)gene is a kind of flowering integron gene,which plays an important role in plant flowering.The current study on the FT gene does not distinguish between flower induction and flower development,probably because the two processes are tightly coupled.Therefore,the FT gene has been less studied in the flower induction stage.In order to further study the functional localization of FT genes during ethephon-induced pineapple flowering,in this study two genes were successfully cloned,AcFT1 and AcFT2,from‘Tainong 16’pineapple and their expression pattern was analyzed.【Methods】The pineapple hybrid variety‘Tainong 16’was cultivated in the Pineapple Greenhouse Planting Base of Meiting Village,Chengmai County,Hainan Province.40%ethephon(1600 mg·L^^-1)diluted with 300 times was used to induce pineapple flowering.Samples were taken at 1 h,3 h,6 h,1 d,6 d,10 d,15 d,22 d and 31 d after treatment,including roots,stems,mature and young leaves,shoot tips and other tissues.After the samples were frozen in the liquid nitrogen,they were stored in a refrigerator at-80℃until use.Stem tips or inflorescence treated with ethephon for 0,10,15,31 and 40 days were fixed in FAA fixative solution,and then dehydration,dyeing,transparency,wax dipping,embedding and slicing were carried out.The total RNA was extracted using the BioTeKe polysaccharide polyphenol plant RNA extraction kit,and the cDNA was reverse transcribed by the TaKaRa reverse transcription kit.By searching the pineapple genome database,the sequence information of two FT genes was obtained,and based on this information,primers were designed to amplify the pineapple FT gene cDNA.The sequencing results were searched using the BLAST online software program provided by NCBI;the AcFT gene coding region was analyzed by ORF Finder in NCBI;the protein amount and isoelectric point were predicted by online software ProtParam;the protein secondary structure was analyzed by GOV IV.The NCBI database Blastp was used to search the homologous p

关 键 词：菠萝 FT基因 基因克隆 表达分析 

分 类 号：S668.3[农业科学—果树学]