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利用黄盘诱捕器中桃蚜的一步法TaqMan® RT-qPCR检测甜菜感染的马铃薯卷叶病毒,为病毒黄化病的早期风险评估开辟了新的可能性。

One-step TaqMan® RT-qPCR detection of sugar beet-infecting poleroviruses in Myzus persicae from yellow water pan traps opens up new possibilities for early risk assessment of virus yellows disease.

作者信息

Borgolte Simon, Menzel Wulf, Varrelmann Mark

机构信息

Department of Phytopathology, Institute of Sugar Beet Research, Holtenser Landstraße 77, Göttingen D-37079, Germany.

Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Inhoffenstraße 7 B, Braunschweig D-38124, Germany.

出版信息

J Virol Methods. 2025 Jan;331:115052. doi: 10.1016/j.jviromet.2024.115052. Epub 2024 Oct 16.

Abstract

Virus yellows disease (VY) is a major threat to sugar beet production in Europe. Beet chlorosis virus (BChV) and beet mild yellowing virus (BMYV) are of particular economic importance and are both persistently transmitted by the aphid vector Myzus persicae. As part of integrated pest management strategies, M. persicae influx into sugar beet fields is recorded weekly using yellow water pan traps. To date, only ELISA and RT-PCR assays have been described for BChV and BMYV detection in individual aphids. In this study, we describe for the first time two one-step TaqMan® RT-qPCR assays designed for the specific detection of BChV and BMYV in M. persicae after 7d incubation in water pan trap medium. Both viruses were reproducibly detected in individual aphids. After 7d incubation in trap medium, both viruses were reproducibly detected in individual aphids, as well as in one viruliferous aphid in a pool of 99 non-viruliferous aphids. Significant correlations can be shown between different mixing ratios of viruliferous to non-viruliferous aphids and Ct values of total RNA templates, allowing the percentage of viruliferous aphids in yellow water pan traps to be estimated using a standard curve. The described methodology provides a high sensitivity combined with a high sample throughput and can be used, after evaluation in the field, for practical monitoring, risk modelling and development of decision support systems for VY.

摘要

病毒黄化病(VY)是欧洲甜菜生产的主要威胁。甜菜褪绿病毒(BChV)和甜菜轻度黄化病毒(BMYV)具有特别重要的经济意义,且均由蚜虫介体桃蚜持久性传播。作为综合虫害管理策略的一部分,每周使用黄色水盘诱捕器记录桃蚜流入甜菜田的情况。迄今为止,仅描述了用于检测单个蚜虫中BChV和BMYV的ELISA和RT-PCR检测方法。在本研究中,我们首次描述了两种一步法TaqMan® RT-qPCR检测方法,用于在水盘诱捕器培养基中孵育7天后,特异性检测桃蚜中的BChV和BMYV。两种病毒均可在单个蚜虫中被重复检测到。在诱捕器培养基中孵育7天后,两种病毒在单个蚜虫以及99只无毒蚜虫群体中的一只带毒蚜虫中均可被重复检测到。带毒蚜虫与无毒蚜虫的不同混合比例与总RNA模板的Ct值之间存在显著相关性,从而可以使用标准曲线估计黄色水盘诱捕器中带毒蚜虫的百分比。所描述的方法具有高灵敏度和高样本通量,经过田间评估后,可用于实际监测、风险建模以及开发VY的决策支持系统。

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