广东地区冬瓜感染的小西葫芦黄花叶病毒检测及其外壳蛋白基因多样性分析

发布时间：2018-02-21 20:05

  本文关键词： 冬瓜 小西葫芦黄花叶病毒 外壳蛋白基因 遗传变异　出处：《园艺学报》2016年01期 　论文类型：期刊论文

【摘要】：小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)是危害广东省冬瓜的常见病毒。摸索了冬瓜叶片总RNA最佳提取方法,通过基于ZYMV病毒外壳蛋白基因(coat protein gene,cp)序列的RT-PCR扩增,检测采集自广东省9个冬瓜产区的76份病毒侵染样品的病毒田间发病率;将扩增产物克隆测序,利用MegAlign软件构建系统发育进化树分析cp基因遗传变异与系统进化。结果表明,24份样品检测为阳性,田间发病率为31.59%;所有参试病毒分离物cp基因全长840 bp,编码279个氨基酸的多肽;测序的16个分离物cp基因的核苷酸序列与推导蛋白的氨基酸序列相似性分别为98.1%~100%与96.1%~100%;所有广东分离物的推导氨基酸序列基序保守,除GD121-9增加2个人类白细胞抗原外,其他均包含5个人类白细胞抗原及1个N糖基化位点,均包含保守potyv_CP功能域及与病毒蚜传相关的结构域DAG三联盒"Asp-Ala-Gly";系统进化分析表明,ZYMV病毒划分为7个基因型,广东分离物均属于同一基因型Ⅰ,包含3个株系;广东地区的16个分离物分为3组,无明显地域选择性。总体上广东ZYMV-cp比较保守,分子变异较小。明确了广东省冬瓜上ZYMV病毒的侵染情况及外壳蛋白基因的变异特点,为对其致病性和抗病毒基因工程等研究提供依据。
[Abstract]:Zucchini yellow mosaic virus (ZYMV) is a common virus that harms winter melon in Guangdong province. The best method for extracting total RNA from leaves of Chinese winter melon was explored, and the RT-PCR amplification was carried out based on the coat protein gene gene of ZYMV virus. The incidence of virus infection from 76 samples collected from 9 areas of winter melon production in Guangdong Province was detected, and the amplified products were cloned and sequenced. Phylogenetic tree was constructed with MegAlign software to analyze the genetic variation and phylogenetic variation of CP gene. The results showed that 24 samples were positive and the incidence was 31.59.The total length of CP gene was 840 BP, encoding 279 amino acid peptides. The similarity of nucleotide sequence of cp gene and deduced protein was 98.1100% and 96.1%, respectively. The deduced amino acid motifs of all Guangdong isolates were conserved, except for the addition of 2 human leukocyte antigens to GD121-9. The rest contained five human leukocyte antigens and one N-glycosylation site, all of which contained conserved potyv_CP functional domain and DAG triple box "Asp-Ala-Gly" associated with virus aphid transmission. Phylogenetic analysis showed that the virus was divided into seven genotypes. Guangdong isolates belong to the same genotype 鈪，

本文编号：1522758