Complete genome sequencing and infectious cDNA clone construction of a Chinese isolate of grapevine Pinot gris virus (GPGV)

IF 2.2 4区农林科学 Q2 PLANT SCIENCES

Journal of Plant Pathology Pub Date : 2024-08-13 DOI:10.1007/s42161-024-01737-6

Qingyun Yuan, Ying Zhang, Fang Ren, Guojun Hu, Xudong Fan, Yafeng Dong

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Abstract

The complete nucleotide sequence of a Chinese isolate C12B1 of grapevine Pinot gris virus (GPGV) from a ‘Beta’ grapevine was determined by small RNA sequencing. The size of the C12B1 genome was 7259 nucleotides (nt) without the poly (A) tail. Sequence identities of C12B1 with other GPGV isolates ranged from 79.13 to 97.52%. Phylogenetic analysis based on complete genome sequences of the global GPGV isolates indicated GPGV isolates are divided into three major groups and five subgroups within group 3, with the C12B1 isolate felled within subgroup 3a of group 3. Moreover, an infectious cDNA clone of the GPGV isolate C12B1 was generated using the seamless assembly approach. The infectious clone was then inoculated to Nicotiana occidentalis 37B and grapevine plants. Inoculated Nicotiana plants showed leaf chlorotic mottling symptoms and RT-PCR and transmission electron microscopy analysis confirmed successful infection. Additionally, the inoculated Beta grapevine plants also exhibited mild chlorotic mottling and ring spot symptoms. This study represents the first complete genome sequence of GPGV in China and the successful production of an infectious cDNA clone of a divergent GPGV isolate. These findings are a foundation for further research on the interaction between GPGV and its plant hosts.

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葡萄灰比诺病毒（GPGV）中国分离株的全基因组测序和感染性 cDNA 克隆构建

通过小核糖核酸（small RNA）测序，确定了从 "Beta "葡萄树上分离的中国葡萄灰比诺病毒（GPGV）C12B1的完整核苷酸序列。C12B1 基因组的大小为 7259 个核苷酸（nt），不含聚 (A) 尾。C12B1 与其他 GPGV 分离物的序列相同度为 79.13% 至 97.52%。根据全球 GPGV 分离物的完整基因组序列进行的系统进化分析表明，GPGV 分离物分为 3 大类，在第 3 类中有 5 个亚类，C12B1 分离物属于第 3 类中的 3a 亚类。此外，利用无缝组装法生成了 GPGV C12B1 分离物的感染性 cDNA 克隆。然后将该感染性克隆接种到西洋烟草 37B 和葡萄植株上。接种的烟草植株出现叶片萎黄斑驳症状，RT-PCR 和透射电子显微镜分析证实感染成功。此外，接种的 Beta 葡萄植株也出现了轻微的叶片萎黄斑驳和环斑症状。本研究首次在中国完成了 GPGV 的基因组测序，并成功制备了一个具有传染性的 GPGV 分离物 cDNA 克隆。这些发现为进一步研究 GPGV 与植物宿主之间的相互作用奠定了基础。

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来源期刊

Journal of Plant Pathology 生物-植物科学

CiteScore

3.10

自引率

4.50%

发文量

218

审稿时长

6-12 weeks

期刊介绍： The Journal of Plant Pathology (JPP or JPPY) is the main publication of the Italian Society of Plant Pathology (SiPAV), and publishes original contributions in the form of full-length papers, short communications, disease notes, and review articles on mycology, bacteriology, virology, phytoplasmatology, physiological plant pathology, plant-pathogeninteractions, post-harvest diseases, non-infectious diseases, and plant protection. In vivo results are required for plant protection submissions. Varietal trials for disease resistance and gene mapping are not published in the journal unless such findings are already employed in the context of strategic approaches for disease management. However, studies identifying actual genes involved in virulence are pertinent to thescope of the Journal and may be submitted. The journal highlights particularly timely or novel contributions in its Editors’ choice section, to appear at the beginning of each volume. Surveys for diseases or pathogens should be submitted as "Short communications".