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比利时的甜菜土传苹果褪绿叶斑病毒及其与根肿病的关系。

The Beet soilborne pomovirus in Belgium and relationship with rhizomania.

作者信息

Stas A, Meunier A, Schmit J F, Marlier A, Steyer S, Bragard C

机构信息

Unité de phytopathologie, Faculté des sciences agronomiques, Université catholique de Louvain, Place Croix du Sud, 2bte 3, B-1348 Louvain-la-Neuve.

出版信息

Meded Rijksuniv Gent Fak Landbouwkd Toegep Biol Wet. 2001;66(2a):39-45.

Abstract

The objective of this study was to determine the extend of the Beet soilborne pomovirus (BSBV) and the Beet virus Q in sugar beet fields in Belgium. During the 2000 sugar beet growing season, more than 80 fields located in Belgium were investigated for the presence of the Beet necrotic yellow vein benyvirus (BNYVV), the BSBV and Polymyxa betae, the plasmodiophorid vector of both viruses. Fields investigated were identified either using previous recorded data or by visual identification of yellow leaves on sugar beets or root symptoms. Sampling position (longitude-latitude) was recorded using the global positioning system (G.P.S.) with the view to follow-up infested fields in the following years. Three different techniques were used to evidence the aforementioned biological agents: enzyme-linked immunosorbent assay (ELISA), a RT-PCR assay to detect the viruses and direct coloration of Polymyxa betae in plant root tissues, using lactophenol-aniline blue. ELISA allowed the detection of 43 BSBV-infested soils, largely distributed in all Belgian sugar beet growing areas. These results were largely confirmed by RT-PCR using two different primers pairs targeting respectively a 400 bp fragment of the 145K ORF located on virus RNA-1 and a 970 bp fragment of the conserved 3' end of the viral genome. Five other primer's pairs have also been evaluated for BSBV identification. The detection of BSBV-infested soils without BNYVV, as well as BNYVV-infested soils without BSBV allowed the design of a competition assay between both viruses. Among the samples, 21 were selected randomly and tested for the presence of Beet virus Q by RT-PCR. Here also, six fields were detected positive for this virus. Sequence data reveal a clonal population of BSBV isolates in Belgium though a high level of diversity is observed for the coat protein region. Compared to BSBV, BVQ isolates show less diversity at sequence level.

摘要

本研究的目的是确定比利时甜菜田中的甜菜土壤传播多面体病毒(BSBV)和甜菜Q病毒的分布范围。在2000年甜菜生长季节,对比利时80多个甜菜田进行了调查,以检测甜菜坏死黄脉病毒(BNYVV)、BSBV以及两种病毒的传播介体甜菜多黏菌。所调查的田块通过先前记录的数据或通过目视识别甜菜上的黄叶或根部症状来确定。使用全球定位系统(GPS)记录采样位置(经度-纬度),以便在接下来的几年中跟踪受侵染的田块。采用三种不同技术来检测上述生物因子:酶联免疫吸附测定(ELISA)、用于检测病毒的逆转录聚合酶链反应(RT-PCR)测定以及使用乳酚-苯胺蓝对植物根组织中的甜菜多黏菌进行直接染色。ELISA检测到43块受BSBV侵染的土壤,广泛分布于比利时所有甜菜种植区。使用分别靶向病毒RNA-1上145K开放阅读框的400 bp片段和病毒基因组保守3'端的970 bp片段的两对不同引物进行RT-PCR,很大程度上证实了这些结果。还评估了另外五对引物用于BSBV鉴定。对不含BNYVV的受BSBV侵染土壤以及不含BSBV的受BNYVV侵染土壤的检测使得能够设计两种病毒之间的竞争试验。在这些样本中,随机选择21个并通过RT-PCR检测甜菜Q病毒的存在。同样,有6块田块检测到该病毒呈阳性。序列数据显示比利时的BSBV分离株为克隆群体,不过在外壳蛋白区域观察到高度的多样性。与BSBV相比,BVQ分离株在序列水平上的多样性较低。

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