A sensitive real-time RT-PCR reveals a high incidence of Southern tomato virus (STV) in Spanish tomato crops

[EN] Southern tomato virus (STV) is a double-stranded RNA (dsRNA) virus belonging to genus Amalgavirus (family Amalgamaviridae). STV has been detected in tomato plants showing different symptoms although it has not been demonstrated that STV is the causal agent. To study the STV incidence and its pathogenic role, a sensitive and quantitative real-time reverse transcription-polymerase chain reaction assay (RT-qPCR) was developed. The standard curve perfonned with viral RNA transcripts allowed a wide dynamic range for STV quantitation from 10(4) to 10(11) copies/ng of total RNA. STV detection by RT-qPCR was 10(2)-fold more sensitive than conventional RT-PCR or RT-LAMP and 10(4)-fold more sens…

A comparative study of viral infectivity, accumulation and symptoms induced by broad bean wilt virus 1 isolates

Broad bean wilt virus 1 (BBWV-1, genus Fabavirus, family Secoviridae) is a bipartite positive-sense single-stranded RNA virus distributed worldwide infecting many herbaceous species. Until now, scarce information regarding biological properties of BBWV-1 isolates is available. This work shows a comparative study on virus infectivity (proportion of infected plants over inoculated plants), virus accumulation and symptoms induced by four genetically different BBWV-1 isolates (Ben, B41/99, NSRV and PV0548) which were mechanically inoculated on several herbaceous hosts. The four BBWV-1 isolates infected broad bean, tomato, pepper and Nicotiana benthamiana plants, whereas none of them infected cu…

Detection of Southern tomato virus by molecular hybridisation

RNA2-encoded VP37 protein of Broad bean wilt virus 1 is a determinant of pathogenicity, host susceptibility, and a suppressor of post-transcriptional gene silencing

Abstract Broad bean wilt virus 1 (BBWV‐1, genus Fabavirus, family Secoviridae) is a bipartite, single‐stranded positive‐sense RNA virus infecting many horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in viral replication whereas RNA2 encodes two coat proteins (the large and small coat proteins) and two putative movement proteins (MPs) of different sizes with overlapping C‐terminal regions. In this work, we determined the role played by the small putative BBWV‐1 MP (VP37) on virus pathogenicity, host specificity, and suppression of post‐transcriptional gene silencing (PTGS). We engineered a BBWV‐1 35S‐driven full‐length cDNA infectious clone corresponding to BBWV‐…

Broad bean wilt virus 1 encoded VP47 and SCP are suppressors of plant post-transcriptional gene silencing

Broad bean wilt virus 1 (BBWV-1, genus Fabavirus, family Secoviridae) is a bipartite positive single-stranded RNA (+ssRNA) virus infecting important horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in virus replication, whereas RNA2 encodes the large and small coat proteins (LCP, and SCP, respectively) and two putative movement proteins with overlapping C-terminal but different sizes: 47.2 kDa (VP47) and 37 kDa (VP37). Post-transcriptional gene silencing (PTGS) is a mechanism of gene regulation and defense against pathogens such as viruses. However, most plant viruses encode proteins called viral suppressors of RNA silencing (VSRs) which able to inhibit PTGS. Pre…

Fast detection of Southern tomato virus by one-step transcription loop-mediated isothermal amplification (RT-LAMP)

Southern tomato virus (STV) is a double stranded RNA (dsRNA) virus belonging to genus Amalgavirus (family Amalgamaviridae) which has been detected in tomato plants showing stunting, fruit discoloration and size reduction. A one-step reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of STV in total RNA or sap extracts (obtained just by grinding in buffer) from STV-infected tomato plants by using a set of three primers pairs which were designed to the sequence of the STV putative coat protein. Amplification products were visualized by gel electrophoresis or direct staining of DNA. The sensitivity of RT-LAMP was identical to that of th…