阮城城;年宇薇;胡福初;王祥和;范鸿雁;吴凤芝;陈哲;张治礼;

• 1:海南大学热带作物学院
• 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
• 3:农业部海口热带果树科学观测实验站
• 4:海南大学生命科学与药学院

摘要(Abstract)：

【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。

关键词(KeyWords)： 菠萝;FT基因;基因克隆;表达分析

Abstract:

Keywords:

基金项目(Foundation): 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

作者(Author): 阮城城;年宇薇;胡福初;王祥和;范鸿雁;吴凤芝;陈哲;张治礼;

Email:

DOI: 10.13925/j.cnki.gsxb.20190170

参考文献(References)：

• [1]刘海清，刘志颐.中国菠萝鲜果产业的国际竞争力分析[J].江苏农业科学，2017,45(17):334-337.LIU Haiqing,LIU Zhiyi. Analysis of international competitiveness of chinese pineapple fresh fruit industry[J]. Jiangsu Agricultural Sciences,2017,45(17):334-337.
• [2]张治礼，范鸿雁，华敏，何凡.菠萝开花诱导及其生理与分子基础[J].热带作物学报，2012,33(5):950-955.ZHANG Zhili,FAN Hongyan,HUA Min,HE Fan. Flowering induction of pineapple and its physiological and molecular basis[J]. Journal of Tropical Crops,2012,33(5):950-955.
• [3]周琴，张思思，包满珠，刘国锋.高等植物成花诱导的分子机理研究进展[J].分子植物育种，2018,16(11):3681-3692.ZHOU Qin,ZHANG Sisi,BAO Manzhu,LIU Guofeng. Advances in molecular mechanisms of flowering induction in higher plants[J]. Molecular Plant Breeding,2018,16(11):3681-3692.
• [4]李宪利，袁志友，高东升.高等植物成花分子机理研究现状及展望[J].西北植物学报，2002,22(1):173-183.LI Xianli,YUAN Zhiyou,GAO Dongsheng. Research status and prospect of molecular mechanisms of flowering in higher plants[J]. Journal of Northwest Botany,2002,22(1):173-183.
• [5] THOMAS J. Molecular and Genetic Mechanisms of Floral Control[J]. The Plant Cell,2004,16:S1-S17.
• [6] YANT L,MATHIEU J,DINH T T,OTT F,LANZ C,WOLLMANN H,CHEN X M,SCHMID M. Orchestration of the floral transition and floral development in Arabidopsis by the bifunctional transcription factor APETALA2[J]. The Plant Cell,2010,22(7):2156-2170.
• [7] CAZZONELLI C I,CAVALLARO A S,BOTELLA J R. Cloning and characterisation of ripening-induced ethylene biosynthetic genes from non-climacteric pineapple(Ananas comosus)fruits[J]. Functional Plant Biology,1998,25(5):513-518.
• [8] CAZZONELLI C I,CAVALLARO A S,BOTELLA J R. Towards the production of transgenic pineapple to control flowering and ripening[J]. Acta Horticulturae,2000,529:115-122.
• [9] WOHRMANN T,WEISING K. In silico mining for simple sequence repeat loci in a pineapple expressed sequence tag database and cross-species amplification of EST-SSR markers across Bromeliaceae[J]. Theoretical&Applied Genetics,2011,123(4):635-647.
• [10] ONG W D,VOO C L Y,KUMAR S V. Development of ESTs and data mining of pineapple EST-SSRs[J]. Molecular Biology Reports,2012,39(5):5889-5896.
• [11] LV L L,DUAN J,XIE J H,LIU Y G,WEI C B,LIU S H,ZHANG J X,SUN G M. Cloning and expression analysis of a PISTILLATA homologous gene from pineapple(Ananas comosus L. Merr.)[J]. International Journal of Molecular Sciences,2012,13(12):1039-1053.
• [12] MOYLE R L,KOIA J H,VREBALOV J,GIOVANNONI J,BOTELLA J R. The pineapple AcMADS1 promoter confers high level expression in tomato and Arabidopsis flowering and fruiting tissues,but AcMADS1 does not complement the tomato LeMADS-RIN(rin)mutant[J]. Plant Molecular Biology,2014,86(4/5):395-407.
• [13]杨祥燕，蔡元保，张治礼，孙光明，孙伟生，吴青松，王运儒.菠萝MADS-box基因AcMADS2的克隆与表达分析[J].植物生理学报，2016,52(9):1406-1412.YANG Xiangyan,CAI Yuanbao,ZHANG Zhili,SUN Guangming,SUN Weisheng,WU Qingsong,WANG Yunru. Cloning and expression analysis of pineapple MADS-box gene AcMADS2[J]. Journal of Plant Physiology,2016,52(9):1406-1412.
• [14]年宇薇，陈哲，胡福初，范鸿雁，何凡，陈惠萍，张治礼.菠萝LEAFY基因克隆与表达模式研究[J].分子植物育种，2018,16(7):2107-2115.NIAN Yuwei,CHEN Zhe,HU Fuchu,FAN Hongyan,HE Fan,CHEN Huiping,ZHANG Zhili. Cloning and expression pattern of LEAFY gene in pineapple[J]. Molecular Plant Breeding,2018,16(7):2107-2115.
• [15] TANAKA N,URESHINO A,SHIGETA N,MIMIDA N,KOMORI S,TAKAHASHI S,TANAKA-MORIYA Y,WADA M.Overexpression of Arabidopsis FT gene in apple leads to perpetual flowering[J]. Plant Biotechnology,2014,31(1):11-20.
• [16]李球红，费元，庞基良.植物开花整合子基因FT的研究进展[J].北方园艺，2014(18):210-213.LI Qiuhong,FEI Yuan,PANG Jiliang. Advances in the study of flowering integron gene FT in plants[J]. Northern Horticulture,2014(18):210-213.
• [17] SRIKANTH A,SCHMID M. Regulation of flowering time:all roads lead to Rome[J]. Cellular&Molecular Life Sciences,2011,68(12):2013-2037.
• [18]华敏，王祥和，何凡，范鸿雁，何舒，罗革彬.台农16号菠萝催花试验[J].中国南方果树，2009,38(4):49-51.HUA Min,WANG Xianghe,HE Fan,FAN Hongyan,HE Shu,LUO Gebin. Experiments on flowering promotion of pineapple'Tainong 16'[J]. South China Fruits,2009,38(4):49-51.
• [19]杨艺，王娜，王奎玲，刘庆华，刘庆超.铁线莲属4种植物的花芽分化研究[J].园艺学报，2019,46(1):87-95.YANG Yi,WANG Na,WANG Kuiling,LIU Qinghua,LIU Qingchao. Flower bud differentiation of four species of Clematis[J].Acta Horticulturae Sinica,2019,46(1):87-95.
• [20]李瑞雪，余三淼，李夏，熊颖媛，蔺珍，唐琰琪，马均.红苞凤梨实时荧光定量PCR分析中内参基因的筛选[J].热带亚热带植物学报，2017,25(3):250-256.LI Ruixue,YU Sanmiao,LI Xia,XIONG Yingyuan,LIN Zhen,TANG Yanqi,MA Jun. Screening of internal reference genes in real-time fluorescence quantitative PCR analysis of red-bracted pineapple[J]. Journal of Tropical and Subtropical Botany,2017,25(3):250-256.
• [21] LIVAK K J,SCHMITTGEN T D. Analysis of relative gene expression data using realtime quantitative PCR and the 2-ΔΔCtmethod[J]. Methods,2001,25(4):402-408.
• [22] IMAIZUMI T,KAY S A. Photoperiodic control of flowering:not only by coincidence[J]. Trends in Plant Science,2006,11(11):550-558.
• [23] JAEGER K E,GRAF A,WIGGE P A. The control of flowering in time and space[J]. Journal of Experimental Botany,2006,57(13):3415-3418.
• [24] LV L L,DUAN J,XIE J H,WEI C B,LIU Y G,LIU S H,SUN G M. Isolation and characterization of a FLOWERING LOCUS T homolog from pineapple[Ananas comosus(L.)Merr.][J].Gene,2012,505(2):368-373.
• [25] CUNHA G A P D. Applied aspects of pineapple flowering[J].Bragantia,2005,64(4):499-516.
• [26] CHARDON F,DAMERVAL C. Phylogenomic analysis of the PEBP gene family in cereals[J]. Journal of Molecular Evolution,2005,61(5):579-590.
• [27] HEDMAN H,K?LLMAN T,LAGERCRANTZ U. Early evolution of the MFT-like gene family in plants[J]. Plant Molecular Biology,2009,70(4):359-369.
• [28] KARLGREN A,GYLLENSTRAND N,K?LLMAN T,SUNDSTR?M J F,DAVID M,MARTIN L,LAGERCRANTZ U. Evolution of the PEBP gene family in plants:functional diversification in seed plant evolution[J]. Plant Physiology,2011,156:1967-1977.
• [29]朱岩，彭振英，张斌，毕玉平. PEBP家族基因在植物中功能的研究进展[J].山东农业科学，2013,45(2):139-145.ZHU Yan,PENG Zhenying,ZHANG Bin,BI Yuping. Advances in the functions of PEBP family genes in plants[J]. Shandong Agricultural Sciences,2013,45(2):139-145.
• [30]孙洪波，贾贞，韩天富. PEBP家族基因在植物发育调控中的作用[J].植物生理学报，2009,45(8):739-747.SUN Hongbo,JIA Zhen,HAN Tianfu. The role of PEBP family genes in plant development regulation[J]. Journal of Plant Physiology,2009,5(8):739-747.
• [31] YAN L,FU D,LI C,BLECHL A,TRANQUILLI G,BONAFEDE M,SANCHEZ A,VALARIK M,YASUDA S,DUBCOVSKY J. The wheat and barley vernalization gene VRN3 is an orthologue of FT[J]. PNAS,2006,103(51):19581-19586.
• [32] CAI Y P,CHEN L,LIU X J,GUO C,SUN S,WU C X,JIANG B J,HAN T F,HOU W S. CRISPR/Cas9-mediated targeted mutagenesis of GmFT2a delays flowering time in soybean[J]. Plant Biotechnology Journal,2017,16(1):176-185.
• [33] LV B,NITCHER R,HAN X L,WANG S Y,NI F,LI K,PEARCE S,WU J J,DUBCOVSKY J,FU D L. Characterization of FLOWERING LOCUS T1(FT1)gene in Brachypodium and wheat[J]. PLoS One,2014,9(4):e94171.
• [34] KOBAYASHI Y,KAYA H,GOTO K,IWABUCHI M,ARAKI T. A pair of related genes with antagonistic roles in mediating flowering signals[J]. Science,1999,286(5446):1960-1962.
• [35] PIN P A,NILSSON O. The multifaceted roles of FLOWERING LOCUS T in plant development[J]. Plant Cell&Environment,2012,35(10):1742-1755.
• [36] HARIG L,BEINECKE F A,OLTMANNS J,MUTH J,MüLLER O,RüPING B,TWYMAN R M,FISCHER R,PRüFER D,NOLL G. Proteins from the FLOWERING LOCUS T-like subclade of the PEBP family act antagonistically to regulate floral initiation in tobacco[J]. The Plant Journal,2012,72(6):908-921.
• [37] D'ALOIA M,BONHOMME D,BOUCHéF,TAMSEDDAK K,ORMENESE S,TORTI S,COUPLAND G,PéRILLEUX C.Cytokinin promotes flowering of Arabidopsis via transcriptional activation of the FT paralogue TSF[J]. Plant Journal,2011,65(6):972-979.
• [38] NAVARRO C,CRUZ-ORóE,PRAT S. Conserved function of FLOWERING LOCUS T(FT)homologues as signals for storage organ differentiation[J]. Current Opinion in Plant Biology,2015,23:44-53.
• [39]李翔，侯璐，康亚璇，庞晓明，李颖岳.'冬枣'FT同源基因克隆及表达分析[J].果树学报，2017,34(11):1374-1384.LI Xiang,HOU Lu,KANG Yaxuan,PANG Xiaoming,LI Yingyue. Cloning and expression analysis of FT homologous gene of'Dongzao'[J]. Journal of Fruit Science,2017,34(11):1374-1384.