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园艺学报 ›› 2015, Vol. 42 ›› Issue (7): 1347-1355.doi: 10.16420/j.issn.0513-353x.2015-0106

• 观赏植物 • 上一篇    下一篇

菊花开花抑制基因CmFLC-like1的克隆及表达特性分析

展妍丽,王萃铂,亓钰莹,陈发棣,蒋甲福   

  1. 南京农业大学园艺学院,南京 210095
  • 出版日期:2015-07-25 发布日期:2015-07-25
  • 基金资助:
    国家自然科学基金项目(31372100);教育部新世纪优秀人才支持计划项目(NCET-12-0890);江苏省“青蓝工程”和“双创计划”项目

Cloning and Expression Analysis of Flowering Inhibitor CmFLC-like1 Gene in Chrysanthemum

ZHAN Yan-Li, WANG Cui-Bo, QI Yu-Ying, CHEN Fa-Di, JIANG Jia-Fu   

  1. College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
  • Online:2015-07-25 Published:2015-07-25

摘要: 为深入研究夏菊的春化机理,利用多聚酶链式反应(PCR)结合5′ RACE、3′ RACE技术克隆夏菊品种‘优香’[Chrysanthemum morflorium(Ramat.)Kitam.‘Yuuka’]开花抑制基因CmFLC-like1的cDNA全长序列,获得其全长为945 bp,开放阅读框(ORF)为636 bp,编码1条211个氨基酸残基的多肽,且具有典型的MADS结构域。同源性分析表明,CmFLC-like1与葡萄(Vitis vinifera)VvFLC和龙眼(Dimocarpus longan)DlFLC的同源性最高,分别为55%和52%。进化树聚类分析表明,CmFLC-like1蛋白与拟南芥和核桃的FLC遗传距离最近。亚细胞定位表明,CmFLC-like1基因定位在核上。在酵母体系中发现CmFLC-like1没有转录激活活性,拟南芥原生质体转化发现具有转录抑制活性。菊花植株营养生长期不同组织器官的RT-PCR表明CmFLC-like1在叶片中表达量最高,茎和茎尖中次之,根中最少。低温(4 ℃)可抑制CmFLC-like1表达,且处理时间越长,抑制表达越明显。

关键词: 菊花, 春化作用, FLC, RT-PCR

Abstract: The full-length cDNA sequence of Chrysanthemum morflorium(Ramat.)Kitam.‘Yuuka’CmFLC-like1 gene was obtained using PCR and RACE. The cDNA was 945 bp in length with a 636 bp open reading frame which encodes a peptide of 211 residues. CmFLC-like1 has a high homology to VvFLC(Vitis vinifera)and DlFLC(Dimocarpus longan)contained a typical MADS domain,with the homology of 55% and 52%,respectively. CmFLC-like1 was located in the nuclear genome by subcellular localization. And it has no transcription activation activity in yeast system,while it show inhibitory activity in the plastid transformation system. CmFLC-like1 has the highest expression levels in leaves,while the lowest expression in stems and shoot tip,followed by root. The expression of CmFLC-like1 was inhibited by low temperature,especially at 4 ℃. Furthermore,the longer the cold treatment time,the more suppresser ofCmFLC-like1 expression.

Key words: chrysanthemum, vernalization, FLC, RT-PCR

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