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园艺学报 ›› 2020, Vol. 47 ›› Issue (1): 120-126.doi: 10.16420/j.issn.0513-353x.2019-0166

• 研究报告 • 上一篇    下一篇

四川地区猕猴桃病毒1的RT-PCR检测及外壳蛋白基因序列分析

彭期定,吕 蕊,杨 婷,林宏辉,席德慧*   

  1. 四川大学生命科学学院,生物资源与生态环境教育部重点实验室,成都 610065
  • 出版日期:2020-01-25 发布日期:2020-01-25
  • 基金资助:
    国家自然科学基金项目(31270290,31772131);四川省应用基础项目(2019YJ0135);中央高校基础研究经费项目(SCU2019D013)

RT-PCR Detection and Sequence Analysis of Coat Protein Gene of Actinidia virus 1 from Sichuan Province

PENG Qiding,Lü Rui,YANG Ting,LIN Honghui,and XI Dehui*   

  1. Key Laboratory of Bio-resource and Eco-environment of Ministry of Education,College of Life Sciences,Sichuan University,Chengdu 610065,China
  • Online:2020-01-25 Published:2020-01-25

摘要: 为了明确近期新西兰报道的侵染猕猴桃的新病毒——猕猴桃病毒1(Actinidia virus 1,AcV-1)在四川地区猕猴桃上的发生情况及其分子特性,采用RT-PCR对来自四川6个地区疑似感染病毒的90份猕猴桃主栽品种‘红阳’和‘金果’的叶片样品进行检测。结果表明,22份样品为AcV-1阳性,检出率为24.4%。将获得的5个AcV-1分离物和已报道的新西兰分离物K75的外壳蛋白(coat protein,CP)基因序列进行比对,结果表明,分离物间核苷酸序列和氨基酸序列的相似性分别为84.8% ~ 97.1%和89.7% ~ 99.6%,其中除邛崃分离物HYH5与新西兰分离物K75的核苷酸序列相似性较高(97.1%)外,其余4个分离物均低于91%。系统进化分析结果显示,这些分离物主要聚集在3个分支上,分离物HYH5与新西兰分离物K75位于同一分支,其余分离物位于另外2个分支。

关键词: 猕猴桃, 猕猴桃病毒1, RT-PCR, CP基因, 分子特征

Abstract: Recently,a new virus that infected kiwifruit,Actinidia virus 1(AcV-1),has been reported in New Zealand. In order to determine the occurrence and molecular characterization of AcV-1 in kiwifruit cultivars(‘Hongyang’and‘Jinguo’)in Sichuan Province,90 kiwifruit leaves samples of suspected viral infection were tested for AcV-1 by reverse transcription polymerase chain reaction(RT-PCR). The results showed that 22 samples were positive to AcV-1,and the infection rate was 24.4%. The coat protein gene(CP)sequences of the obtained five AcV-1 isolates were compared with isolate K75 which was reported in New Zealand. Results showed that the sequences identities of CP nucleotides and amino acid among these isolates were 84.8%–97.1% and 89.7%–99.6%,respectively. Among them,the sequence identities of CP nucleotides were less than 91% between New Zealand isolate K75 and four of these isolates except Qionglai isolate HYH5,which was 97.1%. Phylogenetic analysis based on CP gene of each isolate showed that these isolates were mainly clustered into three branches,HYH5 and New Zealand isolate K75 were located in the same branch,and the other isolates were located in the other two branches.

Key words: kiwifruit, Actinidia virus 1, RT-PCR, CP gene, molecular characterization

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