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小 RNA-NGS 技术揭示了匈牙利甜樱桃中存在樱桃僵化病毒 A 和李矮缩病毒 1。

Small RNA NGS Revealed the Presence of Cherry Virus A and Little Cherry Virus 1 on Apricots in Hungary.

机构信息

Agricultural Biotechnology Institute, NARIC, 2100 Gödöllő, Hungary.

Department of Biotechnology, Nanophagetherapy Center, Enviroinvest Corporation, 7632 Pécs, Hungary.

出版信息

Viruses. 2018 Jun 11;10(6):318. doi: 10.3390/v10060318.

Abstract

Fruit trees, such as apricot trees, are constantly exposed to the attack of viruses. As they are propagated in a vegetative way, this risk is present not only in the field, where they remain for decades, but also during their propagation. Metagenomic diagnostic methods, based on next generation sequencing (NGS), offer unique possibilities to reveal all the present pathogens in the investigated sample. Using NGS of small RNAs, a special field of these techniques, we tested leaf samples of different varieties of apricot originating from an isolator house or open field stock nursery. As a result, we identified Cherry virus A (CVA) and little cherry virus 1 (LChV-1) for the first time in Hungary. The NGS results were validated by RT-PCR and also by Northern blot in the case of CVA. Cloned and Sanger sequenced viral-specific PCR products enabled us to investigate their phylogenetic relationships. However, since these pathogens have not been described in our country before, their role in symptom development and modification during co-infection with other viruses requires further investigation.

摘要

果树,如杏树,会不断受到病毒的侵袭。由于它们是通过营养繁殖方式传播的,因此这种风险不仅存在于它们在田间保留几十年的过程中,也存在于其繁殖过程中。基于下一代测序(NGS)的宏基因组诊断方法为揭示研究样本中所有现有病原体提供了独特的可能性。我们使用小 RNA 的 NGS 技术,即这些技术的一个特殊领域,测试了来自隔离室或开放田间苗圃的不同品种杏叶的样本。结果,我们首次在匈牙利鉴定出樱桃病毒 A(CVA)和小樱桃病毒 1(LChV-1)。NGS 结果通过 RT-PCR 进行了验证,对于 CVA 还通过 Northern blot 进行了验证。克隆和 Sanger 测序的病毒特异性 PCR 产物使我们能够研究它们的系统发育关系。然而,由于这些病原体在我国以前没有被描述过,因此它们在与其他病毒共同感染时对症状发展和改变的作用需要进一步研究。

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