Cereal viruses – brome mosaic virus: prevalence and diagnostics

Virus-like particles (VLPs), due to their nanoscale dimensions, presence of interior cavities, self-organization abilities and responsiveness to environmental changes, are of interest in the field of nanotechnology. Nevertheless, comprehensive knowledge of VLP self-assembly principles is incomplete. VLP formation is governed by two types of interactions: protein–cargo and protein–protein. These interactions can be modulated by the physicochemical properties of the surroundings. Here, we used brome mosaic virus (BMV) capsid protein produced in an E. coli expression system to study the impact of ionic strength, pH and encapsulated cargo on the assembly of VLPs and their features. We showed that empty VLP assembly strongly depends on pH whereas ionic strength of the buffer plays secondary but significant role. Comparison of VLPs containing tRNA and polystyrene sulfonic acid (PSS) revealed that the structured tRNA profoundly increases VLPs stability. We also designed and produced mutated BMV capsid proteins that formed VLPs showing altered diameters and stability compared to VLPs composed of unmodified proteins. We also observed that VLPs containing unstructured polyelectrolyte (PSS) adopt compact but not necessarily more stable structures. Thus, our methodology of VLP production allows for obtaining different VLP variants and their adjustment to the incorporated cargo.

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

Molecular Biology Reports ◽

10.1007/s11033-021-06485-9 ◽

2021 ◽

Author(s):

Katarzyna Trzmiel

Keyword(s):

Mosaic Virus ◽

Simultaneous Detection ◽

Brome Mosaic Virus ◽

Mottle Virus ◽

Grass Species ◽

Replicase Gene ◽

Rt Pcr ◽

Pcr Products ◽

Cereal Plants ◽

Cocksfoot Mottle Virus

AbstractBrome mosaic virus (BMV) and cocksfoot mottle virus (CfMV) are pathogens of grass species including all economically important cereals. Both viruses have been identified in Poland therefore they create a potential risk to cereal crops. In this study, a duplex—reverse transcription—polymerase chain reaction (duplex-RT-PCR) was developed and optimized for simultaneous detection and differentiation of BMV and CfMV as well as for confirmation of their co-infection. Selected primers CfMVdiag-F/CfMVdiag-R and BMV2-F/BMV2-R amplified 390 bp and 798 bp RT-PCR products within coat protein (CP) region of CfMV and replicase gene of BMV, respectively. Duplex-RT-PCR was successfully applied for the detection of CfMV-P1 and different Polish BMV isolates. Moreover, one sample was found to be co-infected with BMV-ML1 and CfMV-ML1 isolates. The specificity of generated RT-PCR products was verified by sequencing. Duplex-RT-PCR, like conventional RT-PCR, was able to detect two viruses occurring in plant tissues in very low concentration (as low as 4.5 pg/µL of total RNA). In contrast to existing methods, newly developed technique offers a significant time and cost-saving advantage. In conclusion, duplex-RT-PCR is a useful tool which can be implemented by phytosanitary services to rapid detection and differentiation of BMV and CfMV.

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Coding Changes in the 3a Cell-to-Cell Movement Gene Can Extend the Host Range of Brome Mosaic Virus Systemic Infection

Virology ◽

10.1006/viro.1995.0057 ◽

1995 ◽

Vol 214 (2) ◽

pp. 464-474 ◽

Cited By ~ 24

Author(s):

WALTER DE JONG ◽

ANTONY CHU ◽

PAUL AHLQUIST

Keyword(s):

Host Range ◽

Mosaic Virus ◽

Cell Movement ◽

Systemic Infection ◽

Brome Mosaic Virus

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CRYO-EM Atomic Model of Brome Mosaic Virus Derived from Direct Electron Detection Images and a Real-Space Model Optimization Protocol

Biophysical Journal ◽

10.1016/j.bpj.2013.11.3321 ◽

2014 ◽

Vol 106 (2) ◽

pp. 600a ◽

Cited By ~ 1

Author(s):

Zhao Wang ◽

Corey Hryc ◽

Benjamin Bammes ◽

Pavel Afonine ◽

Joanita Jakana ◽

...

Keyword(s):

Mosaic Virus ◽

Real Space ◽

Brome Mosaic Virus ◽

Atomic Model ◽

Model Optimization ◽

Space Model ◽

Electron Detection

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cDNA cloning and in vitro transcription of the complete brome mosaic virus genome

Molecular and Cellular Biology ◽

10.1128/mcb.4.12.2876-2882.1984 ◽

1984 ◽

Vol 4 (12) ◽

pp. 2876-2882 ◽

Cited By ~ 1

Author(s):

P Ahlquist ◽

M Janda

Keyword(s):

Mosaic Virus ◽

Transcription Initiation ◽

Direct Sequencing ◽

In Vitro Transcription ◽

Brome Mosaic Virus ◽

Viral Rna ◽

In Vitro Translation ◽

In Vitro Production ◽

Genomic Rnas

Complete cDNA copies of each of the brome mosaic virus genomic RNAs (3.2, 2.8, and 2.1 kilobases in length) were cloned in a novel transcription vector, pPM1, designed to provide exact control of the transcription initiation site. After cleavage at a unique EcoRI site immediately downstream of the inserted cDNA, these clones can be transcribed in vitro by Escherichia coli RNA polymerase to yield complete copies of the brome mosaic virus RNAs. Dideoxy sequencing of 5' transcript cDNA runoff products and direct sequencing of 32P-3'-end-labeled transcripts show that such transcripts initiate at the same 5' position as natural viral RNA and terminate within the EcoRI runoff site after copying the entire viral RNA sequence. When synthesized in the presence of m7GpppG, the transcripts bear the natural capped 5' terminus of brome mosaic virus RNAs. Such transcripts direct the in vitro translation of proteins which coelectrophorese with the translation products of natural brome mosaic virus RNAs. pPM1 should facilitate in vitro production of other viral and nonviral RNAs.

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