Molecular Characterization and Genomic Function of Grapevine Geminivirus A
Molecular Characterization and Genomic Function of Grapevine Geminivirus A
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葡萄双生病毒 A 的分子特征和基因组功能
DOI：
10.3389/fmicb.2020.555194
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发表时间：
2020-09
影响因子：
5.2
通讯作者：
Yajuan Qian
中科院分区：
文献类型：
--
作者：
Suwei Sun;Ya Hu;Guangzhuang Jiang;Yimin Tian;Ming Ding;Cui Yu;Xueping Zhou;Yajuan Qian
采用sRNA高通量测序和传统桑格测序技术，从云南元谋葡萄上分离到一个新的葡萄藤双生病毒A（GGVA）分离物（命名为GGVA-17 YM 1）及其缺陷基因组（GGVA-D）。为了研究GGVA和GGVA-D的致病性，构建了GGVA-17 YM 1和GGVA-D-17 YM 1的感染性克隆。侵染试验表明，GGVA单独或GGVA与GGVA-D组合接种的本氏烟草植株表现出顶叶向上卷曲和矮化。Southern杂交和实时荧光定量聚合酶链反应分析表明，GGVA-D增加了GGVA DNA的积累水平。利用PVX衍生的重组载体进行瞬时表达表明，GGVA编码的C2和C4参与了N.本萨米亚纳此外，V2蛋白在GFP转基因N.本萨米亚那植物随后，全长基因组测序鉴定出11种不同的GGVA分离株和9种相关的缺陷DNA分子。基于全基因组序列的系统发育分析表明，所有GGVA分离株，包括我们的序列，聚类成两个不同的分支，没有地理分组。单核苷酸多态性（SNPs）的分子变异分析表明，转换（55.97%）多于颠换（44.03%）。此外，ORF C1、C3或V1的主要变体是同义突变，ORF C2、C4和V2的主要变体是非同义突变。遗传选择分析表明，V1、C1、C2和C3四个ORF受到负选择作用，V2和C4受到正选择作用。我们的研究结果有助于GGVA的遗传多样性的表征，并提供其致病性的见解。
A new grapevine geminivirus A (GGVA) isolate (named as GGVA-17YM1) and its associated defective genome (GGVA-D) were identified from a grapevine sample collected in Yuanmou, Yunnan Province, using sRNA high throughput sequencing and traditional Sanger sequencing. To explore the pathogenicity of GGVA and GGVA-D, infectious clones of GGVA-17YM1 and GGVA-D-17YM1 were constructed. Infection assays indicated that Nicotiana benthamiana plants inoculated with GGVA alone or a combination of GGVA and GGVA-D exhibited upward curled apical leaves and dwarfism. Southern blotting and quantitative real-time polymerase chain reaction analysis revealed that GGVA-D increased the accumulation level of GGVA DNA. Transient expression using a PVX-derived recombinant vector indicated that C2 and C4 encoded by GGVA are involved in symptom induction in N. benthamiana. Furthermore, the V2 protein inhibited local RNA silencing in co-infiltration assays in GFP transgenic N. benthamiana plants. Subsequently, full-length genome sequencing resulted in the identification of 11 different isolates of GGVA and 9 associated defective DNA molecules. Phylogenetic analysis based on whole genome sequences showed that all GGVA isolates, including our sequences, clustered into two distinct branches with no geographical grouping. Analyses of molecular variation indicated single nucleotide polymorphisms (SNPs) with more transitions (55.97%) than transversions (44.03%). Furthermore, the main variants for ORF C1, C3, or V1 were synonymous mutations, and non-synonymous mutations for ORF C2, C4, and V2. Genetic selection analysis indicated that negative selection acted on four ORFs (V1, C1, C2, and C3), while V2 and C4 were under positive selection. Our results contribute to the characterization of the genetic diversity of GGVA and provide insights into its pathogenicity.
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影响因子：
5.4
作者：
Cui, XF;Tao, XR;Zhou, XP
通讯作者：
Zhou, XP
影响因子：
5.3
作者：
Lima ATM;Silva JCF;Silva FN;Castillo-Urquiza GP;Silva FF;Seah YM;Mizubuti ESG;Duffy S;Zerbini FM
通讯作者：
Zerbini FM
影响因子：
1.4
作者：
L. Mostert;E. Abeln;F. Halleen;P. Crous
通讯作者：
L. Mostert;E. Abeln;F. Halleen;P. Crous
影响因子：
3.8
作者：
Mar, Talita Bernardon;Diniz Xavier, Cesar Augusto;Murilo Zerbini, F.
通讯作者：
Murilo Zerbini, F.
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影响因子：
20.5
作者：
Zeng Runxiu;Liu Xiaoshi;Yang Chengwei;Lai Jianbin
通讯作者：
Lai Jianbin