利用新的分子标记研究俄罗斯联邦不同地区常见的马铃薯病毒 Y 型分离株

A. A. Stakheev, A. I. Uskov, Yu. A. Varitsev, P. A. Galushka, L. B. Uskova, S. V. Zhevora, S. K. Zavriev
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摘要

摘要 本文报告了一项利用新标记序列研究马铃薯病毒 Y 分离物遗传多态性的研究。这项工作于 2022-2023 年进行。分析使用了 2017 年从生长在俄罗斯联邦不同地区的国内外品种中收集的 PVY 分离物,并对其进行了血清型预选。选择了三个位点作为系统发育信息含量较高的潜在标记,即衣壳蛋白基因(Cp)、NTR的5'-非翻译区和VPg蛋白基因,后者与RNA的5'-末端共价结合并作为转录调节因子。NTR 片段的扩增效率最高(100% 扩增测试的 cDNA 样品)。它在系统发育方面的信息量也最大:对所研究序列的比较分析表明,可变位置和系统发育信息量分别为 23.8%和 15%。在对 NTR 标记序列进行比较分析的基础上构建的系统发生树包括几个由高引导值支持的聚类。这些聚类包括来自以下亚群的分离物:PVYNTN-Red Scarlett (III)、Red Scarlett (5-4) 和 Dunya (7-12);PVYN:O-Pomdor、Zagadka 和 Nida;PVYO/C-Picasso、Favorit (III)、CH991 131 (8-2)、Floris (7-14) 和 Romano (5-3);PVYWilga-Red Scarlett (6-10) 和 Kenza,以及属于这些亚群的模式株,其标记序列已存入 NCBI 数据库。从马铃薯品种 Zhukovsky rannii(5-2)中分离出的一个菌株处于坏死群和序群之间的中间位置,在基于 NTR 位点标记核苷酸序列的系统发生树上形成了一个单独的分支,没有与任何类型菌株归为一类。测序分析基本证实了测试样本的初步血清学鉴定,表明坏死和重组菌株群占优势。
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Study of Potato Virus Y Isolates Commonly Occurring in Different Regions of the Russian Federation Using New Molecular Markers

Abstract

The paper reports the study which was carried out to investigate the genetic polymorphism of potato virus Y isolates using new marker sequences. The work was performed in 2022–2023. PVY isolates collected in 2017 from domestic and foreign varieties grown in different regions of the Russian Federation and preselected for serotype were used for analysis. Three loci were selected as potential markers with high phylogenetic information content, namely, the coat protein gene (Cp), the 5'-untranslated region of NTR, and the VPg protein gene, which covalently binds to the 5'-end of RNA and acts as a transcription regulator. Highest amplification efficiency (100% of the cDNA samples tested) was achieved using the NTR fragment. It was also the most phylogenetically informative: a comparative analysis of the studied sequences showed the presence of 23.8 and 15% variable and phylogenetically informative positions, respectively. The phylogenetic tree constructed on the basis of the comparative analysis of the NTR marker sequences included several clusters supported by high bootstrap values. The clusters encompassed the isolates from the following subgroups: PVYNTN—Red Scarlett (III), Red Scarlett (5-4), and Dunya (7-12); PVYN:O—Pomdor, Zagadka, and Nida; PVYO/C—Picasso, Favorit (III), CH991 131 (8-2), Floris (7-14), and Romano (5-3), and PVYWilga—Red Scarlett (6-10) and Kenza, as well as the type strains belonging to these subgroups, the marker sequences of which being deposited in the NCBI database. An isolate from the potato variety Zhukovsky rannii (5-2) showed an intermediate position between the necrotic and ordinal clusters, forming a separate branch and not grouping with any of the type strains on the phylogenetic tree based on the marker nucleotide sequences of the NTR locus. Sequencing analysis generally confirmed the initial serological identification of the tested samples demonstrating the predominance of the necrotic and recombinant groups of strains.

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