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园艺学报 ›› 2014, Vol. 41 ›› Issue (5): 889-906.

• 蔬菜 • 上一篇    下一篇

番茄抗黄化曲叶病毒病相关基因ClNLR 的功能分析

李 楠1,*,杨玛丽1,*,陈天子2,张保龙2,赵统敏1,杨郁文2,刘廷利2,余文贵1,**   

  1. 1 江苏省农业科学院蔬菜研究所,南京 210014;2 江苏省农业科学院,江苏省生物学重点实验室,南京 210014
  • 出版日期:2014-05-25 发布日期:2014-05-25
  • 基金资助:

    江苏省自然科学基金项目(BK2011676);江苏省农业科技自主创新资金项目[CX(11)1003];国家自然科学基金项目(31301775)

Functional Characterization of ClNLR Gene Involved in Tomato Defense
Against Tomato Yellow Leaf Curl Disease

  1. 1Institute of Vegetable Crops,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2Provincial Key Laboratory of Agro-biology,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China
  • Online:2014-05-25 Published:2014-05-25

摘要: 在前期通过转录组研究发现1 个NBS-LRR 类抗病基因(Solyc05g009760.1)在抗TYLCV
番茄材料‘CLN2777A’中上调表达,推测可能参与抵抗TYLCV 侵染的防卫反应的基础上,以‘CLN2777A’
番茄为材料,通过RT-PCR 克隆该基因全长cDNA 序列,命名为ClNLR。荧光定量PCR 发现ClNLR 在番
茄‘CLN2777A’的根和叶片中高水平表达。ClNLR 在本氏烟叶片上的瞬时表达诱导了过敏性反应。病毒
诱导的基因沉默(VIGS)抑制ClNLR 基因在抗病番茄‘CLN2777A’中表达后,接种TYLCV,检测叶片
带毒量,发现VIGS 处理的番茄植株体内的TYLCV 积累量与其ClNLR 基因表达水平成反比。这些研究结
果表明ClNLR 可能为抗番茄黄化曲叶病的相关基因。

关键词: 番茄, 番茄黄化曲叶病毒, NBS-LRR, 基因克隆, 表达分析, 过敏性反应, 基因沉默

Abstract: The previous comparative transcriptome revealed the NBS-LRRs gene(Solyc05g009760.1)
was up-regulated in the TYLCV resistant tomato line CLN2777A and may involve in the resistance to
TYLCV. In this study,the cDNA of this NBS-LRRs gene was isolated from CLN2777A by RT-PCR and
named as ClNLR. Quantative RT-PCR revealed that ClNLR gene was strongly expressed in the root and
leaves of tomato. The transient expression of ClNLR gene in leaves of Nicotiana benthamiana triggered the
hypersensitive reaction(HR). And the virus induced gene silencing(VIGS)of ClNLR gene in the
CLN2777A plantlets facilitated the proliferation of TYLCV as shown by a negative relationship between
the DNA content of TYLCV and the expression levels of ClNLR gene in the VIGS treated plantlets. Taken together,ClNLR gene may be a TYLCV resistance-related gene in tomato.

Key words: tomato, Tomato yellow leaf curl virus, NBS-LRR, gene clone, expression analysis, hypersensitive reaction, gene silence

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