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园艺学报 ›› 2011, Vol. 38 ›› Issue (6): 1129-1138.

• 观赏植物 • 上一篇    下一篇

菊花花发育基因CmCO和CmFT的克隆与表达分析

田素波,林桂玉,郑成淑*,孙 霞,任洪艳,温立柱   

  1. (山东农业大学园艺科学与工程学院,作物生物学国家重点实验室,山东泰安 271018)  
  • 收稿日期:2011-03-09 修回日期:2011-04-30 出版日期:2011-06-25 发布日期:2011-06-25
  • 通讯作者: 郑成淑

Cloning and Expression of CmCO and CmFT of Floral Development Genes in Chrysanthemum

TIAN Su-bo,LIN Gui-yu,ZHENG Cheng-shu*,SUN Xia,REN Hong-yan,and WEN Li-zhu   

  1. (College of Horticulture Science and Engineering,Shandong Agricultural University,State Key Laboratory of Crop Biology,Tai’an,Shandong 271018,China)   
  • Received:2011-03-09 Revised:2011-04-30 Online:2011-06-25 Published:2011-06-25
  • Contact: ZHENG Cheng-shu

摘要:  利用同源序列法结合RACE技术从菊花‘神马’品种[Chrysanthemum morflorium(Ramat.)Kitam.‘Jinba’]中分离了开花时间相关的CO(CONSTANS)和FT(FLOWERING LOCUS T)同源基因,并命名为CmCO(基因登录号JF488070)和CmFT(基因登录号JF488071)。CmCO和CmFT分别编码382和174个氨基酸。蛋白比对发现,CmCO蛋白包含具有典型的CO同源蛋白结构,包含B-box1,B-box2,CCT结构域及COOH区域。CmFT所推测的氨基酸序列包含FT类蛋白保守基序和两个关键性氨基酸残基。同源性分析表明,CmCO与草莓(Fragaria × ananassa)FaCO同源性最高,为65.8%,与豌豆(Pisum sativum)PsCOL、拟南芥(Arabidopsis thaliana)AtCO同源性分别为62.0%和55.6%。CmFT与向日葵(Helianthus annuus)HaFT2基因同源性最高,为93.7%,与葡萄(Vitis vinifera)VvFT和拟南芥AtFT的同源性分别为85.1%和74.0%。进化树聚类分析表明,CmCO和CmFT蛋白分别与向日葵HaCO和HaFT2遗传距离最近。RT-PCR表明,长日照下的菊花叶片中几乎检测不到CmCO和CmFT,而在短日照下,CmCO在花芽分化启动期(Ⅰ)表达,总苞鳞片分化前期(Ⅱ)有所下降随后又迅速升高;CmFT在CmCO之后表达,之后持续高表达。选择小花原基分化前期(Ⅳ)对菊花叶片、花芽和茎等不同组织器官CmCO和CmFT表达进行分析,结果表明,CmCO在叶片中表达量最高,花芽次之,茎最低;CmFT在花芽中表达量最高,叶片次之,茎最低。由此推测CmCO和CmFT的表达与光周期诱导菊花成花密切相关。   

关键词: 菊花, 光周期, 开花, CO, FT

Abstract: The genes of CO(CONSTANS)and FT(FLOWERING LOCUS T),which related to flowering time were isolated from Chrysanthemum morflorium(Ramat.)Kitam.‘Jinba’by RT-PCR and rapid amplification of cDNA ends(RACE),and named it CmCO(GenBank accession No. JF488070)and CmFT(GenBank accession No. JF488071)respectively. CmCO and CmFT encoded 382 and 174 amino acids respectively. Alignment analysis showed that CmCO protein had typical structures of CO homologous protein,including B-box1,B-box2,CCT domain and COOH region. The deduced amino acid sequence of CmFT contained FT-like protein’s conserved motifs and two critical amino acid residues. Homologous analysis showed that the homology of CmCO compared with FaCO(Fragaria × ananassa)was the highest by 65.8%,and with PsCOL(Pisum sativum)and AtCO(Arabidopsis thaliana)were by 62.0% and 55.6% . The homology of CmFT compared with HaFT2(Helianthus annuus)was the highest by 93.7%,and with VvFT(Vitis vinifera)and with AtFT(Arabidopsis thaliana)were 85.1% and 74.0%. Evolutionary tree clustering analysis indicated that the genetic distances between CmCO protein and HaCOL(Helianthus annuus)and HaFT2 were the shortest respectively among others. RT-PCR analysis showed that expression of CmCO and CmFT in the leaves of chrysanthemum almost didn’t observed under long-day conditions,but under short-day conditions,CmCO expressed at initial stage of floral bud differentiation(Ⅰ),then decreased at the stage of involucre primordial diferentiation(Ⅱ)followed by a quick rise. CmFT expressed followed by the CmCO expression,and then kept to high expression. The expression of CmCO and CmFT at the leaves,floral buds and stems of chrysanthemum at initial stage of floret primordial differentiation(Ⅳ)indicated that the expression of CmCO in leaves of chrysanthemum was the highest,and it in the floral buds lower,and it in stems the lowest. The expression of CmFT in the floral buds was the highest,and in the leaves lower,and in stems the lowest. The results presumed that the expressions of CmCO and CmFT were related with flowering of chrysanthemum induced by photoperiods.    

Key words: chrysanthemum, photoperiod, flowering, CO, FT

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