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园艺学报 ›› 2020, Vol. 47 ›› Issue (12): 2405-2414.doi: 10.16420/j.issn.0513-353x.2020-0170

• 研究报告 • 上一篇    下一篇

柑橘CsNCED3-2的克隆及其响应溃疡病菌侵染的表达分析

谢 宇,张庆雯,祁静静,邹修平,何永睿,许兰珍,雷天刚,彭爱红,李 强,姚利晓,陈善春*,龙 琴*   

  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:2020-12-25 发布日期:2021-01-06
  • 基金资助:
    国家重点研发计划项目(2018YFD1000300);重庆市自然科学基金—博士后基金项目(cstc2019jcyj-bshX0024);中央高校基本科研业务费项目(XDJK2019C027);国家现代农业产业技术体系建设专项资金项目(CARS-26)

Cloning and Expression Analysis of CsNCED3-2 in Responding to Citrus Canker Disease

XIE Yu,ZHANG Qingwen,QI Jingjing,ZOU Xiuping,HE Yongrui,XU Lanzhen,LEI Tiangang,PENG Aihong,LI Qiang,YAO Lixiao,CHEN Shanchun*,and LONG Qin*   

  1. Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,National Citrus Engineering Research Center,Chongqing 400712,China
  • Online:2020-12-25 Published:2021-01-06

摘要: 柑橘溃疡病是世界柑橘的一种重要病害,对柑橘产业造成严重的经济损失。对柑橘中9–顺式–环氧类胡萝卜素双加氧酶(NCED)基因家族进行注释,并克隆出受柑橘溃疡病菌诱导表达的CsNCED3-2,确定了其表达模式。从柑橘全基因组公共数据库共注释出14个NCED基因家族成员,根据系统发育树和功能结构域将其分为4个亚类;CsNCED3-2其序列全长2 377 bp,开放阅读框1 830 bp,编码609个氨基酸,属于RPE65超家族成员和类胡萝卜素双加氧酶家族成员,是非分泌性蛋白;在‘晚锦橙’和‘金弹’叶片注射溃疡病菌后的各个时间点,‘晚锦橙’叶片中CsNCED3-2的相对表达量总体高于‘金弹’,并且两品种的相对表达变化呈相反趋势;与茎、叶和种子相比,CsNCED3-2在两品种果皮中表达量较高;两品种的CsNCED3-2上游启动子均含有参与植物激素和逆境应答相关的顺式作用元件ABRE(脱落酸响应)、TCA-element(水杨酸响应)、MYC/MYB(干旱响应)等,但数量和类型存在差异。

关键词: 柑橘, 溃疡病, NCED, 基因克隆, 表达量, 启动子

Abstract: Citrus canker is an important disease of citrus in the world and causes severe economic losses to the citrus industry. In this study,the 9-cis-epoxycarotenoid dioxygenase gene(NCED)family of Citrus sinensis were annotated,and the citrus canker related CsNCED3-2 was cloned and its expression pattern was analyzed. A total of 14 NCED family members were annotated from the public genomic databases of C. sinensis. These 14 NCED family members can be divided into 4 subclasses based on the phylogenetic tree and functional domain. The full-length of CsNCED3-2 is 2 377 bp with a 1 830 bp open reading frame which codes a protein containing 609 amino acids. The CsNCED3-2 gene belongs to the members of the RPE65 superfamily and the carotenoid dioxygenase family,and it is a non-secretory protein. After inoculated with Xanthomonas citri subsp. citri(Xcc),the expression of CsNCED3-2 in Jindan(Fortunella crassifolia Swingle)was always higher than that in Wanjincheng orange(Citrus sinensis Osbeck)at different time points,and the variation trends of expression in the two varieties were opposite. The expression of CsNCED3-2 in the peel of Wanjincheng Orange and Jindan were higher than that in the stem,leaf and seed. The CsNCED3-2 promoters of Wanjincheng Orange and Jindan contain multiple cis-acting elements involved in plant adversity and hormone responses,such as ABRE(abscisic acid response),TCA-element(salicylic acid response),MYC/MYB(drought response)and so on,but there are differences in their number and type.

Key words: citrus, canker disease, NCED, gene cloning, expression level, promoter

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