研究报告
菠萝花发育相关基因AcMADS1的克隆与组织表达特性分析
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- 1广西壮族自治区亚热带作物研究所, 南宁 530001;
2中国热带农业科学院南亚热带作物研究所, 湛江 524091
3广西壮族自治区农业科学院园艺研究所, 南宁 530007;
4海南大学园艺园林学院, 海口 570228
5海南省农业科学院, 海口 571100
收稿日期: 2013-05-07
修回日期: 2013-11-03
网络出版日期: 2014-11-21
基金资助
公益性行业（农业）科研专项
Cloning of Flowering-related Gene AcMADS1 and Characterization of Expression in Tissues of Pineapple (Ananas comosus)
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- 1Guangxi Subtropical Crops Research Institute, Nanning 530001, China;
2South Subtropical Crop Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China;
3Horticultural Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, China;
4College of Horticulture and Landscape Architecture, Hainan University, Haikou 570228, China;
5Hainan Academy of Agricultural Sciences, Haikou 571100, China
Received date: 2013-05-07
Revised date: 2013-11-03
Online published: 2014-11-21
摘要
MADS-box转录因子在植物的发育过程特别是控制花器官的诱导与发育中起关键作用。利用同源克隆结合RACE技术, 从菠萝(Ananas comosus)花中分离出1个新的菠萝MADS-box基因, 命名为AcMADS1(GenBank登录号为KC257408)。AcMADS1基因的编码区为726 bp, 编码241个氨基酸, 蛋白质分子量为27.50 kDa, 等电点为9.26。序列比对和系统进化树分析表明, AcMADS1具有保守的MADS-box及半保守的K区, 属于AGL6亚家族MADS-box蛋白。生物信息学分析表明, AcMADS1是亲水碱性蛋白, 二级结构主要以α-螺旋、无规则卷曲和折叠延伸链为蛋白质骨架, 三级结构中蛋白核心结构符合转录因子与DNA结合的常见功能域MADS-box, 而且作为转录因子定位于细胞核中。组织特异性表达分析表明, AcMADS1基因在菠萝果肉以及花器官的雌蕊、花瓣和萼片中均有表达, 但在雄蕊以及营养器官的根、茎和叶中几乎不表达; 且在花器官早期发育过程中大量表达, 后期呈下降趋势。因此推测这个基因可能在菠萝花器官发育和开花诱导过程中起重要作用。
本文引用格式
蔡元保, 杨祥燕, 孙光明, 黄强, 刘业强, 李绍鹏, 张治礼 . 菠萝花发育相关基因AcMADS1的克隆与组织表达特性分析[J]. 植物学报, 2014 , 49(6) : 692 -703 . DOI: 10.3724/SP.J.1259.2014.00692
Abstract
MADS-box gene is a key transcription factor that plays crucial roles in plant development, especially controlling the formation and development of floral organs. We cloned the flower-specific gene from floral organs of Ananas comosus, named AcMADS1 (GeneBank accession no. KC257408), was cloned by homology-based cloning and RACE. The open reading frame of AcMADS1 was 726 bp, encoding 241 amino acids with molecular weight 27.5 kDa and isoelectric point 9.26. Sequence alignment and phylogenetic tree analysis indicated that the deduced protein AcMADS1 contained a conservative MADS-box and semi-conservative K domain and belonged to the AGL6 clade of the MADS-box family. Bioinformatics analysis demonstrated that AcMADS1 was an alkaline and hydrophilic protein; the secondary structure comprised an α-helix, a random coil and an extended strand; the protein core structure agreed with the transcription factors and the function of the common DNA combining domain MADS-box in its protein 3D model and was located in the nucleus as a transcription factor. Tissue-specific analysis showed that AcMADS1 was expressed in floral organs (pistils, petals and sepals) and flesh of pineapple, with nearly no transcription detected in vegetative organs (roots, stems and leaves) and stamens. Furthermore, the expession of AcMADS1 was higher at the early phase of floral development, then reduced at the late phase. AcMADS1 may have important roles in floral induction and floral development of pineapple.
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[30]Theissen G,Becker A,Di Rosa A,Kanno A,Kim JT,Münster T,Winter KU,Saedler H.A short history of MADS-box genes in plants[J].Plant Mol Biol,2000,42:115-149
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[32]Thompson BE,Bartling L,Whipple C,Hall DH,Sakai H,Schmidt R,Hake S.Bearded-ear encodes a MADS box transcription factor critical for maize floral development[J].Plant Cell,2009,21:2578-2590
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[34]Viaene T,Vekemans D,Becker A,Melzer S,Geuten K.Expression divergence of the AGL6 MADS domain transcription factor lineage after a core eudicot duplication suggests functional diversification.[J].BMC Plant Biol,2010,10:148-
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[36]Yang Y,Fanning L,Jack T.The K domain mediates heterodimerization of the Arabidopsis floral organ identity proteins,APETALA3 and PISTILLATA[J].Plant J,2003,33:47-59
[2]王珍华,胡立霞,钟丹,韩雪,庞基良.AGAMOUS like 6亚家族基因研究进展[J].西北植物学报,2012,32:1480-1487
[3]杨祥燕,蔡元保,吴青松,孙光明.菠萝锌指蛋白基因AcRCHY1的克隆与表达分析[J].园艺学报,2009,36:1589-1596
[4]Angenent GC,Franken J,Busscher M,van Dijken A,van Went JL,Dons HJ,van Tunen AJ.A novel class of MADS box genes is involved in ovule development in petunia[J].Plant Cell,1995,7:1569-1582
[5]Becker A,Thei?en G.The major clades of MADS-box genes and their role in the development and evolution of flowering plants[J].Mol Phylogenet Evol,2003,29:464-489
[6]Boss PK,Sensi E,Hua C,Davies C,Thomas MR.Cloning and characterisation of grapevine (Vitis vinifera L[J].) MADS-box genes expressed during inflorescence and berry development. Plant Sci,2002,162:887-895
[7]Coen ES,Meyerowitz EM.The war of the whorls: genetic interactions controlling flower development[J].Nature,1991,353:31-37
[8]de Folter S,Immink RGH,Kieffer M,Parenicova L,Henz SR,Weigel D,Busscher M,Koolker M,Colombo L,Kater MM,Davies B,Angenent GC.Comprehensive interaction map of the Arabidopsis MADS-box transcription factors[J].Plant Cell,2005,17:1424-1433
[9]Ferrario S,Immink RG,Angenent GC.Conservation and diversity in flower land[J].Curr Opin Plant Biol,2004,7:84-91
[10]Gramzow L,Ritz MS,Thei?en G.On the origin of MADS-domain transcription factors[J].Trends Genet,2010,26:149-153
[11]Hsu HF,Huang CH,Chou LT,Yang CH.Ectopic expression of an orchid (Oncidium Gower Ramsey) AGL6-like gene promotes flowering by activating flowering time genes in Arabidopsis thaliana[J].Plant Cell Physiol,2003,44:783-794
[12]Immink RG,Gadella TWJ,Ferrario S,Busscher M,Angenent GC.Analysis of MADS box protein-protein interactions in living plant cells[J].P Natl Acad Sci USA,2002,99:2416-2421
[13]Irish VF (2003).The evolution of floral homeotic gene function. Bioessays,,25(7): 637–645
[14]Jack T.Molecular and genetic mechanisms of floral control[J].Plant Cell,2004,16:1-17
[15]Kaufmann K,Melzer R,Thei?en G.MIKC-type MADS-domain proteins: structural modularity,protein interactions and network evolution in land plants[J].Gene,2005,347:183-198
[16]Koo SC,Bracko O,Park MS,Schwab R,Chun HJ,Park KM,Seo JS,Grbic V,Balasubramanian S,Schmid M,Godard F,Yun DJ,Lee SY,Cho MJ,Weigel D,Kim MC.Control of lateral organ development and flowering time by the Arabidopsis thaliana MADS-box Gene AGAMOUS-LIKE6[J].Plant J,2010,62:807-816
[17]Li H,Liang W,Jia R,Yin C,Zong J,Kong H,Zhang D.The AGL6-like gene OsMADS6 regulates floral organ and meristem identities in rice[J].Cell Res,2010,20:299-313
[18]Lin EP,Peng HZ,Jin QY,Deng MJ,Li T,Xiao XC,Hua XQ,Wang KH,Bian HW,Han N,Zhu MY.Identification and characterization of two bamboo (Phyllostachys praecox) AP1/SQUA-like MADS-box genes during floral transition[J].Planta,2009,231:109-120
[19]Liu C,Zhang J,Zhang N,Shan H,Su K,Zhang J,Meng Z,Kong H,Chen Z.Interactions among proteins of floral MADS-box genes in basal eudicots: implications for evolution of the regulatory network for flower development[J].Mol Biol Evol,2010,27:1598-1611
[20]Ma H,Yanofsky MF,Meyerowitz EM.AGL1-AGL6,an Arabidopsis gene family with similarity to floral homeotic and transcription factor genes[J].Gene Dev,1991,5:484-495
[21]Melzer R,Wang YQ,Thei?en G.The naked and the dead: The ABC's of gymnosperm reproduction and the origin of the angiosperm flower[J].Semin Cell Dev Biol,2009,21:118-128
[22]Ohmori S,Kimizu M,Sugita M,Miyao A,Hirochika H,Uchida E,Nagato Y,Yoshida H.MOSAIC FLORAL ORGANS1,an AGL6-Like MADS box gene,regulates floral organ identity and meristem fate in rice[J].Plant Cell,2009,21:3008-3025
[23]Parenicová L,de Folter S,Kieffer M,Horner DS,Favalli C,Busscher J,Cook HE,Ingram RM,Kater MM,Davies B,Angenent GC,Colombo L.Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis: new openings to the MADS world[J].Plant Cell,2003,15:1538-1551
[24]Pelaz S,Ditta GS,Baumann E,Wisman E,Yanofsky MF.B and C floral organ identity functions require SEPALLATA MADS-box genes[J].Nature,2000,405:200-203
[25]Reinheimer R,Kellogg EA.Evolution of AGL6-like MADS box genes in grasses (Poaceae): ovule expression is ancient and palea expression is new[J].Plant Cell,2009,21:2591-2605
[26]Rounsley SD,Ditta GS,Yanofsky MF.Diverse roles for MADS box genes in Arabidopsis development[J].Plant Cell,1995,7:1259-1269
[27]Schwarz-Sommer Z,Huijser P,Nacken W,Saedler H,Sommer H.Genetic control of flower development by homeotic genes in Antirrhinum majus[J].Science,1990,250:931-936
[28]Smaczniak C,Immink RG,Angenent GC,Kaufmann K.Developmental and evolutionary diversity of plant MADS-domain factors: insights from recent studies[J].Development,2012,139:3081-3098
[29]Tapia-López R,García-Ponce B,Dubrovsky JG,Garay-Arroyo A,Pérez-Ruíz RV,Kim SH,Acevedo F,Pelaz S,Alvarez-Buylla ER.An AGAMOUS-related MADS-box gene,XAL1 (AGL12),regulates root meristem cell proliferation and flowering transition in Arabidopsis[J].Plant Physiol,2008,146:1182-1192
[30]Theissen G,Becker A,Di Rosa A,Kanno A,Kim JT,Münster T,Winter KU,Saedler H.A short history of MADS-box genes in plants[J].Plant Mol Biol,2000,42:115-149
[31]Thei?en G,Saedler H.Plant biology: Floral quartets[J].Nature,2001,409:469-471
[32]Thompson BE,Bartling L,Whipple C,Hall DH,Sakai H,Schmidt R,Hake S.Bearded-ear encodes a MADS box transcription factor critical for maize floral development[J].Plant Cell,2009,21:2578-2590
[33]Tian B,Chen YY,Yan YX,Li DZ.Isolation and ectopic expression of a bamboo MADS-box gene[J].Chinese Sci Bull,2005,50:217-224
[34]Viaene T,Vekemans D,Becker A,Melzer S,Geuten K.Expression divergence of the AGL6 MADS domain transcription factor lineage after a core eudicot duplication suggests functional diversification.[J].BMC Plant Biol,2010,10:148-
[35]Weigel D,Meyerowitz EM.The ABCs of floral homeotic genes. [J].Cell,1994,78:203-209
[36]Yang Y,Fanning L,Jack T.The K domain mediates heterodimerization of the Arabidopsis floral organ identity proteins,APETALA3 and PISTILLATA[J].Plant J,2003,33:47-59