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园艺学报 ›› 2021, Vol. 48 ›› Issue (5): 1023-1030.doi: 10.16420/j.issn.0513-353x.2020-1004

• 研究报告 • 上一篇    下一篇

橘蚜传播率不同的柑橘衰退病毒分离株CPm的表达特性和分子特征

刘金香1, 王欢欢1, 王洪苏1,2, 周彦1, 李中安1, 周常勇1,*()   

  1. 1西南大学/中国农业科学院柑桔研究所,重庆 400712
    2四川省中医药科学院,成都 610041
  • 收稿日期:2021-01-26 修回日期:2021-03-05 出版日期:2021-05-25 发布日期:2021-06-07
  • 通讯作者: 周常勇 E-mail:zhoucy@cric.cn
  • 基金资助:
    国家自然科学基金项目(31972237);国家重点研发计划项目(2018YFD0201500);中央高校基本科研业务费项目(XDJK2019B017)

Molecular Characteristic and Expression Level of Minor Coat Protein of Citrus Tristeza Virus Isolates with Different Transmissibility by Toxoptera citricida

LIU Jinxiang1, WANG Huanhuan1, WANG Hongsu1,2, ZHOU Yan1, LI Zhongan1, ZHOU Changyong1,*()   

  1. 1Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,Chongqing 400712,China
    2Sichuan Academy of Chinese Medicine Sciences,Chengdu 610041,China
  • Received:2021-01-26 Revised:2021-03-05 Online:2021-05-25 Published:2021-06-07
  • Contact: ZHOU Changyong E-mail:zhoucy@cric.cn

摘要:

柑橘衰退病毒(citrus tristeza virus,CTV)通过媒介昆虫传播必需要有病毒小外壳蛋白(minor coat protein,CPm)的参与。为解析褐色橘蚜传播率不同的CTV分离株CPm的分子特征和表达特性,通过克隆、测序获得其CPm基因序列,将对应的氨基酸序列运用DNAStar软件进行相似性和序列比对分析,并应用Western blot技术分析了3个传播率不同的中国CTV分离株CPm的表达特性。结果表明,低传播率的分离株CTLJ与其他分离株CPm氨基酸序列差异较大,同源性仅为93.3% ~ 94.2%,氨基酸序列中有14个位点发生变异,其中9个位点是该分离株特有的。尤其有些位点的氨基酸极性和电荷发生了变化,引起了蛋白质二级结构的改变。然而,传播率不同的CTV分离株CPm的表达水平却无显著差异。这些结果揭示了CTV被褐色橘蚜传播的效率与其CPm的表达特性无关,可能与其氨基端序列变异有关。

关键词: 柑橘衰退病毒, 小外壳蛋白, 表达特性, 分子特征

Abstract:

The minor coat protein(CPm)of citrus tristeza virus(CTV)is necessary in virus transmission by the brown citrus aphid[Toxoptera citricida(Kirkaldy)]. The expression levels of CPm in three isolates with high,middle and low aphid transmissibility were conducted by Western blot technique. Then,the CPm sequences of CTLJ and CT16-2 isolates were acquired by RT-PCR,cloning and sequencing. Finally,these amino acid sequences were used to analyze the multiple alignments by the DNAStar program. The result showed that the identity of amino acid sequence of CPm for CTLJ with low aphid transmissibility was only 93.3%-94.2%,when compared with those of other CTV isolates. Comparison of the CPm of these isolates showed that nine of fourteen amino acid mutation site were unique in the CTLJ isolate. Particularly,some mutations led to substitution of new amino acids with a different charge and polarity,which resulted in changes of the secondary structure of the CPm protein in CTLJ. However,the expression of CPm for CTLJ,CT16-2 and CT11A did not show a significant difference. The results suggest that the aphid transmission efficiency of CTV might be not related to the expression level of CPm,but might be associated with amino acid site variation.

Key words: citrus tristeza virus(CTV), minor coat protein(CPm), expression level, molecular characteristics

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