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中国农学通报 ›› 2018, Vol. 34 ›› Issue (25): 26-31.doi: 10.11924/j.issn.1000-6850.casb18050155

• 生物技术科学 • 上一篇    下一篇

乌头花叶病病原物的分子鉴定

何静,侯大斌   

  1. 西南科技大学生命科学与工程学院东七E座203,西南科技大学生命科学与工程学院东七E座203
  • 收稿日期:2018-05-30 修回日期:2018-06-14 接受日期:2018-07-03 出版日期:2018-09-07 发布日期:2018-09-07
  • 通讯作者: 侯大斌
  • 基金资助:
    国家自然科学基金“柴胡Aux/IAA 基因对侧根形成的影响”(81603223);四川省科技支撑计划“四川道地中药材附子高产高效栽培关键技 术研究与集成示范”(2015NZ0083) 。

Pathogen Molecular Identification of Mosaic Disease Aconitum

  • Received:2018-05-30 Revised:2018-06-14 Accepted:2018-07-03 Online:2018-09-07 Published:2018-09-07

摘要: 研究旨在通过逆转录聚合酶链式反应(RT-PCR)从分子方面鉴定附子地道产区四川江油乌头花叶病植株的病原物种类及分析其特性。通过对黄瓜花叶病毒(Cucumber mosaic virus, CMV)的外壳蛋白基因(Coat Protein,CP)设计特异引物,对江油中坝附子GAP生产基地的乌头花叶病植株进行克隆、转化并测序。结果表明:从江油花叶病乌头植株中克隆得到基因大小为375 bp的片段,鉴定为黄瓜花叶病毒,命名为CMV-FZ,该序列为一段有效的长为375 nt的氨基酸编码区,推测编码125个氨基酸残基。依据外壳蛋白核苷酸序列建立了CMV不同分离物的系统进化树并对该序列进行蛋白组学分析,CMV-FZ与国内外已报道代表性CMV亚组分离物CP基因的核苷酸与氨基酸的序列相似性为74.8%~98.2%和73.4%~99.2%,其中与与CMV亚组II的核苷酸和氨基酸序列相似性为96.1%-98.2%和96.0%-99.2%,该分离物与中国分离物(CMV-PCT2)氨基酸序列相似性最高,达到99.2%。遗传进化分析显示,CMV-FZ与中国分离物(Zin和Hubei-1)和美国分离物(DKRD)亲缘关系较近,聚在同一分支,属于CMV亚组II。以上结果表明伴有花叶症状的乌头受到黄瓜花叶病毒的侵染。

关键词: 两系不育系, 两系不育系, 强分蘖, 有效穗, 产量

Abstract: The study aims to identify the pathogenic species of the Acoitum mosaic plant from Jiangyou, Sichuan Province, which is the local production area of Aconitum, and analyze the characteristics of the pathogenic species by polymerase chain reaction (RT-PCR) in respect of molecular. Specific primers of coat protein (cp) of cucumber mosaic virus (CMV) from Zhongba Jiangyou were designed, then cloned, transformed and sequenced. A 375 bp gene fragment was identified as CMV and named CMV- FZ. The sequence of 375 gene fragment was an effective code region of amino acid, coding 125 amino acid residues. The phylogenetic tree of different CMV isolates was established by the coat protein nucleotide sequence, and then the proteomics was analyzed for this sequence. Compared with the study result of CMV virus subgroup isolates cp gene, the sequence similarity was 74.8%-98.2% and 73.4%-99.2% for nucleotide and amino acid respectively, and the similarity to CMV virus subgroup II was 96.1%-98.2% and 96.0%-99.2% respectively. CMV- FZ embraced the highest amino sequence similarity with CMV-PCT2 from China, up to 99.2%. Genetic evolution analysis showed that the affinity of CMV-FZ, Chinese isolates (Zin and Hubei-1) and American isolates (DKRD) was the closest, they were together in the same branch of the phylogenetic tree, belonged to the CMV subgroup II. The above results exhibited that Acontium accompanied with mosaic disease was infected with CMV.