Detection of infectious Brome mosaic virus in irrigation ditches and draining strands in Poland

Abstract

Environmental waters, e.g. rivers, lakes and irrigation water, are a good source of many plant viruses. The pathogens can infect plants getting through damaged root hairs or small wounds that appear during plant growth. First results demonstrated common incidence of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in water samples collected from irrigation ditches and drainage canals surrounding fields in Southern Greater Poland. Principal objective of this work was to examine if environmental water might be the source of viruses infective to cereals. The investigation was focused on mechanically transmitted pathogens. Virus identification was performed by biological, electron microscopic, serological and molecular methods. Preliminary assays demonstrated Brome mosaic virus (BMV) infections in symptomatic plants inoculated with 9 out of the 17 tested concentrated water samples. The final identification was confirmed by molecular methods for selected isolates named: BMV-Ch1, -DBS, -N, -R and -S. Partial coding sequences of polymerase 1a (RNA1) and 2a (RNA2) and complete nucleotide sequence of coat protein (CP) gene (RNA3) of each BMV isolate were determined and compared with corresponding sequences of other known BMV isolates.Results confirmed the highest amino acid sequence homology in the fragment of polymerase 2a (99.2% -100%) and the most divergence in CP (96.2% -100%). This is the first report on the detection of an infective cereal virus in aqueous environment.

Duplex-RT-PCR assay for the simultaneous detection and discrimination of Brome mosaic virus and Cocksfoot mottle virus in cereal plants

“…Based on published results [2,13] BMV and CfMV can be a new emerging threat for cereal crops. To date CfMV has been detected only in one locations in Poland [11] however, since first detection of BMV in 1999 in Poland, its presence has been reported in different regions of the country [5,8].…”

“…Taking into account the possibility of further spread of the studied pathogens, the designing of specific diagnostic technique as the first step in the successful management of the diseases is needed. Existing diagnostic methods for BMV [2,5,8,[17][18][19] and CfMV [11,[13][14][15][16] have their limitations. ELISA is a time-consuming and laborious technique while conventional RT-PCR assay is able to detect only individual virus species in a single reaction.…”

“…Since both viruses have morphologically identical spherical virions, are transmitted by the same vectors, and induce similar symptoms on infected plants, proper diagnostics of these viruses can be problematic. To date, detection of BMV and CfMV is based on biological tests, enzyme-linked immunosorbent assays (ELISA) or reverse-transcription-polymerase chain reactions (RT-PCR) [2,5,8,[11][12][13][14][15][16][17][18][19]. A multiplex-reverse transcription-polymerase chain reaction (multiplex-RT-PCR) is the promising alternative for above mentioned techniques which has been developed for the rapid and efficient detection of many RNA viruses in one reaction [5,20,21].…”

Aspects in Tobamovirus Management in Intensive Agriculture

“…Infectious tobamovirus particles of TMV and ToMV were found in environmental waters such as ponds and streams. The occurrence of the tobamoviruses was visualized by electron microscopy, and the infectivity of the tobamovirus particles was examined in a biological assay on laboratory test plants [69]. A quantitative and sensitive approach to detect the tobamoviruses in environmental waters was also applied using sensitive reverse-transcription real-time PCR analysis [70].…”

Inactivation of Pepper Mild Mottle Virus in Water by Cold Atmospheric Plasma

“…Another tobamovirus, cucumber green mottle mosaic virus (CGMMV), is also re-emerging and causing significant losses of cucurbit crops ( Dombrovsky et al, 2017 ). Reports of infective tobamoviruses in treated wastewater ( Bačnik et al, 2020 ) or environmental waters ( Jeżewska et al, 2018 ) and their survival in the nutrient solution for up to 6 months ( Pares et al, 1992 ) suggest that water might have an important role in the epidemiology and transmission of these viruses. This has been further confirmed by studies that have reported water-mediated transmission of CGMMV ( Li et al, 2016 ) and other tobamoviruses, and this becomes especially relevant for crops grown using hydroponics ( Mehle and Ravnikar, 2012 ).…”