Zygocactus virus X-based expression vectors and formation of rod-shaped virus-like particles in plants by the expressed coat proteins of Beet necrotic yellow vein virus and Soil-borne cereal mosaic virus

Koenig, R.; Lesemann, D.-E.; Loss, Scott R.; Engelmann, J.; Commandeur, Ulrich; Deml, G.; Schiemann, Joachim; Aust, Hans-Jürgen; Burgermeister, W.

doi:10.1099/vir.0.81477-0

Cited by 7 publications

(3 citation statements)

References 23 publications

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“…Stability has been improved in other vectors with duplicated SGPs by using heterologous promoters to reduce the likelihood of recombination [59,63,65,66]. Accordingly, we generated a hybrid vector containing the SGPs from two distantly related PepMV isolates, PepMV-Sp13 and PepMV-Ps5.…”

Section: Discussionmentioning

confidence: 99%

Development of expression vectors based on pepino mosaic virus

et al. 2011

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BackgroundPlant viruses are useful expression vectors because they can mount systemic infections allowing large amounts of recombinant protein to be produced rapidly in differentiated plant tissues. Pepino mosaic virus (PepMV) (genus Potexvirus, family Flexiviridae), a widespread plant virus, is a promising candidate expression vector for plants because of its high level of accumulation in its hosts and the absence of severe infection symptoms. We report here the construction of a stable and efficient expression vector for plants based on PepMV.ResultsAgroinfectious clones were produced from two different PepMV genotypes (European and Chilean), and these were able to initiate typical PepMV infections. We explored several strategies for vector development including coat protein (CP) replacement, duplication of the CP subgenomic promoter (SGP) and the creation of a fusion protein using the foot-and-mouth disease virus (FMDV) 2A catalytic peptide. We found that CP replacement vectors were unable to move systemically and that vectors with duplicated SGPs (even heterologous SGPs) suffered from significant transgene instability. The fusion protein incorporating the FMDV 2A catalytic peptide gave by far the best results, maintaining stability through serial passages and allowing the accumulation of GFP to 0.2-0.4 g per kg of leaf tissue. The possible use of PepMV as a virus-induced gene silencing vector to study gene function was also demonstrated. Protocols for the use of this vector are described.ConclusionsA stable PepMV vector was generated by expressing the transgene as a CP fusion using the sequence encoding the foot-and-mouth disease virus (FMDV) 2A catalytic peptide to separate them. We have generated a novel tool for the expression of recombinant proteins in plants and for the functional analysis of virus and plant genes. Our experiments have also highlighted virus requirements for replication in single cells as well as intercellular and long-distance movement.

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Section: Discussionmentioning

confidence: 99%

Development of expression vectors based on pepino mosaic virus

et al. 2011

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“…Subsequently, many potexviruses have been engineered as expression vectors, including bamboo mosaic virus (BaMV) (Chen et al, 2017 ; Lin et al, 1996 ; Liou et al, 2014 ), Cymbidium mosaic virus (CymMV) (Hsieh et al, 2013 ), cassava common mosaic virus (CsCMV) (Tuo et al, 2021 ), Plantago asiatica mosaic virus (PlAMV) (Minato et al, 2014 ), narcissus mosaic virus (NMV) (Zhang et al, 2013 ), Alternanthera mosaic virus (AltMV) (Lim et al, 2010 ), Zygocactus X virus (ZVX) (Koenig et al, 2006 ), pepino mosaic virus (PepMV) (Abrahamian et al, 2021 ; Sempere et al, 2011 ), and FoMV (Bouton et al, 2018 ; Liu et al, 2016 ; Mei et al, 2016 , 2019 ). Of these nine potexvirus species, only FoMV, BaMV, PlAMV, and CymMV can infect monocot plants.…”

Section: Engineering Fomv For Vigs ...mentioning

confidence: 99%

Foxtail mosaic virus: A tool for gene function analysis in maize and other monocots

Beernink

Whitham

2023

Molecular Plant Pathology

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Many plant viruses have been engineered into vectors for use in functional genomics studies, expression of heterologous proteins, and, most recently, gene editing applications. The use of viral vectors overcomes bottlenecks associated with mutagenesis and transgenesis approaches often implemented for analysis of gene function. There are several engineered viruses that are demonstrated or suggested to be useful in maize through proof‐of‐concept studies. However, foxtail mosaic virus (FoMV), which has a relatively broad host range, is emerging as a particularly useful virus for gene function studies in maize and other monocot crop or weed species. A few clones of FoMV have been independently engineered, and they have different features and capabilities for virus‐induced gene silencing (VIGS) and virus‐mediated overexpression (VOX) of proteins. In addition, FoMV can be used to deliver functional guide RNAs in maize and other plants expressing the Cas9 protein, demonstrating its potential utility in virus‐induced gene editing applications. There is a growing number of studies in which FoMV vectors are being applied for VIGS or VOX in maize and the vast majority of these are related to maize–microbe interactions. In this review, we highlight the biology and engineering of FoMV as well as its applications in maize–microbe interactions and more broadly in the context of the monocot functional genomics toolbox.

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“…As noted above, our original infectious clones of AltMV differed in symptom severity and relative levels of replication, based on differences in the RdRp and TGB1 sequences, with a single residue difference in TGB1 having major effects on the efficiency of RNA silencing suppression activity. The CP subgenomic RNA (sgRNA) promoter was identified by comparison to those of other potexviruses (Skryabin et al, 1988;Lee et al, 2000;Koenig et al, 2006) and to the region upstream of the TGB1 start codon; the TGB1 and CP sgRNA promoters were subsequently also determined for portulaca strain AltMV-MU (Putlaev et al, 2015). We modified the infectious clones by duplication of the CP sgRNA promoter, and insertion of a multiple cloning site (MCS) between the Triple Gene Block and the CP gene, with initial insertion of either eGFP or DsRed as fluorescent marker proteins; these initial infectious clones required in vitro transcription of infectious transcripts using the T7 RNA polymerase (Lim et al, 2010a).…”

Section: Utility As a Viral Vectormentioning

confidence: 99%

Alternanthera mosaic virus - an alternative ‘model’ potexvirus of broad relevance

Kim¹,

Lim²

2017

Korean Journal of Agricultural Science

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Alternanthera mosaic virus (AltMV) is a member of the genus Potexvirus which has been known for less than twenty years, and has been detected in Australasia, Europe, North and South America, and Asia. The natural host range to date includes species in at least twentyfour taxonomically diverse plant families, with species in at least four other families known to be infected experimentally. AltMV has been shown to differ from Potato virus X (PVX), the type member of the genus Potexvirus, in a number of ways, including the subcellular localization of the Triple Gene Block 3 (TGB3) protein and apparent absence of interactions between TGB3 and TGB2. Differences between AltMV variants have allowed identification of viral determinants of pathogenicity, and identification of residues involved in interactions with host proteins. Infectious clones of AltMV differing significantly in symptom severity and efficiency of RNA silencing suppression have been produced, suitable either for high level protein expression (with efficient RNA silencing suppression) or for Virus-Induced Gene Silencing (VIGS; with weaker RNA silencing suppression), demonstrating a range of utility not available with most other plant viral vectors. The difference in silencing suppression efficiency was shown to be due to a single amino acid residue substitution in TGB1, and to differences in subcellular localization of TGB1 to the nucleus and nucleolus. The current state of knowledge of AltMV biology, including host range, strain differentiation, host interactions, and utility as a plant viral vector for both protein expression and VIGS are summarized.

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Zygocactus virus X-based expression vectors and formation of rod-shaped virus-like particles in plants by the expressed coat proteins of Beet necrotic yellow vein virus and Soil-borne cereal mosaic virus

Cited by 7 publications

References 23 publications

Development of expression vectors based on pepino mosaic virus

Development of expression vectors based on pepino mosaic virus

Foxtail mosaic virus: A tool for gene function analysis in maize and other monocots

Alternanthera mosaic virus - an alternative ‘model’ potexvirus of broad relevance

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