https://www.ahs.ac.cn/images/0513-353X/images/top-banner1.jpg|#|苹果
https://www.ahs.ac.cn/images/0513-353X/images/top-banner2.jpg|#|甘蓝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner3.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner4.jpg|#|灵芝
https://www.ahs.ac.cn/images/0513-353X/images/top-banner5.jpg|#|桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner6.jpg|#|黄瓜
https://www.ahs.ac.cn/images/0513-353X/images/top-banner7.jpg|#|蝴蝶兰
https://www.ahs.ac.cn/images/0513-353X/images/top-banner8.jpg|#|樱桃
https://www.ahs.ac.cn/images/0513-353X/images/top-banner9.jpg|#|观赏荷花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner10.jpg|#|菊花
https://www.ahs.ac.cn/images/0513-353X/images/top-banner11.jpg|#|月季
https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2017, Vol. 44 ›› Issue (8): 1579-1588.doi: 10.16420/j.issn.0513-353x.2017-0165

• 研究报告 • 上一篇    下一篇

柑橘黄化脉明病毒的TGB基因生物信息学分析及亚细胞定位

崔甜甜 王艳娇 李中安 周常勇 宋 震   

  1. (西南大学/中国农业科学院柑桔研究所,国家柑橘工程技术研究中心,重庆 400712)
  • 出版日期:2017-08-25 发布日期:2017-08-25

Bioinformatics and Subcellular Localization on Citrus yellow vein clearing virus-Triple Gene Block

CUI Tiantian,WANG Yanjiao,LI Zhongan,ZHOU Changyong,and SONG Zhen*   

  1. (Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences;National Citrus Engineering Research Center,Chongqing 400712,China)
  • Online:2017-08-25 Published:2017-08-25

摘要: 柑橘黄化脉明病毒(Citrus yellow vein clearing virus,CYVCV)是一种正义单链RNA病毒。其基因组含有6个开放阅读框(ORF),其中ORF2、ORF3和ORF4组成了三基因连锁结构(triple gene block,TGB)。以感染CYVCV的尤力克柠檬[Citrus limon(L.)Burm. f.]植株的总RNA为模板,扩增和克隆了CYVCV的TGB基因,并开展了其编码蛋白的理化性质和分子特性等生物信息学分析;进而通过In-Fusion?技术构建了TGB各基因的亚细胞定位载体,经农杆菌介导转化洋葱表皮细胞并在荧光显微镜下进行亚细胞定位观察。生物信息学分析结果表明,CYVCV-TGB具有与Potexvirus属病毒TGB相似的理化性质和分子特性,可能参与病毒在寄主中的运动且需要外壳蛋白的参与。亚细胞定位结果显示,TGBp1定位于细胞壁(膜)上,TGBp2和TGBp3主要定位于细胞壁(膜),少量呈星点状定位于细胞内。研究结果为揭示CYVCV在寄主中的运动机理奠定了基础。

关键词: 柑橘, 柑橘黄化脉明病毒, 生物信息学分析, 三基因连锁结构, 亚细胞定位

Abstract: Citrus yellow vein cleansing virus(CYVCV)is a simple positive-sense RNA virus. Its genome contains six open reading frames(ORFs),among which ORF2,ORF3 and ORF4 constitute a triple gene block(TGB). The TGB gene of CYVCV was amplified and cloned by using the total RNA of infected Eureka lemon[Citrus limon(L.)Burm. F.]. Bioinformatic analysis of TGB encoded proteins were carried out with the physicochemical property and molecular characteristics. The TGB gene subcellular localization vector were constructed through the In-Fusion? technology and transformated to onion epidermal cells mediated by Agrobacterium tumefaciens. Subcellular localization was then observed under fluorescence microscope. Bioinformatics analysis results showed that the physicochemical properties and molecular characteristics of CYVCV-TGB were similar to that of the Potexvirus,and the TGB may involved in virus movement in the host. Subcellular localization results showed that TGBp1 was located on the cell wall(cell membrane),while TGBp2 and TGBp3 were mainly located on the cell wall(cell membrane),with a small amount located in the cell. These results laid the foundation of revealing the CYVCV movement mechanism.

Key words: citrus, Citrus yellow vein clearing virus, bioinformatics analysis, triple gene block;subcellular localization

中图分类号: