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园艺学报 ›› 2020, Vol. 47 ›› Issue (5): 817-826.doi: 10.16420/j.issn.0513-353x.2019-0625

• 研究论文 •    下一篇

柑橘超量表达CsNBS-LRR通过SA信号途径增强对溃疡病抗性

李 强,祁静静,窦万福,秦秀娟,何永睿*,陈善春*   

  1. 西南大学/中国农业科学院柑桔研究所,国家柑桔工程技术研究中心,重庆 400712
  • 出版日期:2020-05-25 发布日期:2020-05-25
  • 基金资助:
    国家重点研发计划项目(2018YFD1000306);中央高校基本科研业务费项目(SWU115025);广西科技重大专项(桂科AA18118046-6);国家现代农业产业技术体系建设专项资金项目(CARS-26);重庆市社会事业与民生体系保障科技创新专项(cstc2016shms-ztzx80001,cstc2017shms-xdny80051)

Overexpression of CsNBS-LRR in Citrus Confers Bacterial Canker Resistance by Regulating SA Signaling Pathway

LI Qiang,QI Jingjing,DOU Wanfu,QIN Xiujuan,HE Yongrui*,and CHEN Shanchun*   

  1. Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,National Citrus Engineering Technology Research Center,Chongqing 400712,China
  • Online:2020-05-25 Published:2020-05-25

摘要: 克隆了柑橘CsNBS-LRR基因,分析该基因在柑橘溃疡病菌(Xcc)、水杨酸、茉莉酸甲酯诱导下的表达特征,通过在晚锦橙中超量表达验证其功能。结果表明,CsNBS-LRR的开放阅读框为3 633 bp,编码1 210个氨基酸。其在感病品种晚锦橙中受Xcc诱导下调表达,而在低感品种四季橘中上调表达;在四季橘中受水杨酸诱导明显上调表达,而晚锦橙中上调幅度较小;两品种CsNBS-LRR受茉莉酸甲酯诱导均不明显;超量表达载体转化晚锦橙获得4个阳性植株;抗性评价表明超量表达CsNBS-LRR明显提升了转基因植株对柑橘溃疡病的相对抗性(病情指数为野生型的75% ~ 88%);转基因植株中水杨酸含量和水杨酸合成途径中关键基因转录水平明显提高;水杨酸信号途径中的PR基因转录水平也升高。综上所述,CsNBS-LRR是柑橘抗溃疡病的1个抗病基因,它可能通过对水杨酸途径的调控一定程度上增强柑橘对溃疡病抗性。

关键词: 甜橙, 柑橘溃疡病, CsNBS-LRR, 水杨酸, PR基因

Abstract: The citrus CsNBS-LRR gene was cloned and analyzed for its expression profiles induced by citrus canker and phytohormones. The function was verified by over-expression in Wanjincheng(Citrus sinensis),and further analysis was carried out for the mechanisms. The results showed that the open reading frame of CsNBS-LRR was 3 633 bp,encoding 1 210 amino acids. It was down-regulated by Xanthomonas citri subsp. citri(Xcc)in the susceptible variety Wanjincheng,and up-regulated in the resistant variety Calamondin(Citrus madurensis). In Calamondin,CsNBS-LRR was significantly up-regulated by salicylic acid(SA),while in Wanjincheng it was slightly increased. In the two varieties,CsNBS-LRR was not significantly induced by methyl jasmonate(MeJA). Overexpression(OE)of CsNBS-LRR significantly results in significantly enhanced resistance to citrus bacterial canker(CBC)in the transgenic lines,as revealed by the disease severity which was 75%–88% of the wild type. The salicylicacid content in the transgenic plants and the transcription level of key genes in the salicylic acid biosynthesis were significantly increased. Transcriptional levels of key genes in salicylic acid signaling pathway are also enhanced and more sensitive to the Xcc infection. Over all,CsNBS-LRR is a potential disease resistance gene of CBC,which can improve CBC resistance through the regulation of salicylic acid signaling pathway.

Key words: sweet orange, citrus bacterial canker(CBC), CsNBS-LRR, salicylic acid, PR gene

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