Development and optimization of a pepino mosaic virus-based vector for rapid expression of heterologous proteins in plants

Abrahamian, Peter; Hammond, Rosemarie W.

doi:10.1007/s00253-020-11066-0

Cited by 5 publications

(4 citation statements)

References 58 publications

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“…However, when this vector was used to express the GFP-encoding gene, only weak GFP fluorescence and expression were observed in the systemic leaves of cassava plants infected with CsCMV-GFP. Various strategies have been used to enhance the expression of heterologous proteins in plants using potexvirus-based vectors, including optimizing the length of the SGP, replacing the SGP with sequences from related poxviruses, incorporating gene expression enhancer elements, and mutating viral genes to promote virus accumulation [ 34 , 39 , 43 – 45 ]. Previous studies using FoMV-mediated VOX vectors have shown that increasing the length of the duplicated SGP2 sequence can enhance the expression of heterologous proteins [ 34 ].…”

Section: Discussionmentioning

confidence: 99%

Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava

Tuo

Yao

et al. 2023

Plant Methods

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Background Plant virus vectors designed for virus-mediated protein overexpression (VOX), virus-induced gene silencing (VIGS), and genome editing (VIGE) provide rapid and cost-effective tools for functional genomics studies, biotechnology applications and genome modification in plants. We previously reported that a cassava common mosaic virus (CsCMV, genus Potexvirus)-based VIGS vector was used for rapid gene function analysis in cassava. However, there are no VOX and VIGE vectors available in cassava. Results In this study, we developed an efficient VOX vector (CsCMV2-NC) for cassava by modifying the CsCMV-based VIGS vector. Specifically, the length of the duplicated putative subgenomic promoter (SGP1) of the CsCMV CP gene was increased to improve heterologous protein expression in cassava plants. The modified CsCMV2-NC-based VOX vector was engineered to express genes encoding green fluorescent protein (GFP), bacterial phytoene synthase (crtB), and Xanthomonas axonopodis pv. manihotis (Xam) type III effector XopAO1 for viral infection tracking, carotenoid biofortification and Xam virulence effector identification in cassava. In addition, we used CsCMV2-NC to deliver single guide RNAs (gMePDS1/2) targeting two loci of the cassava phytoene desaturase gene (MePDS) in Cas9-overexpressing transgenic cassava lines. The CsCMV-gMePDS1/2 efficiently induced deletion mutations of the targeted MePDS with the albino phenotypes in systemically infected cassava leaves. Conclusions Our results provide a useful tool for rapid and efficient heterologous protein expression and guide RNA delivery in cassava. This expands the potential applications of CsCMV-based vector in gene function studies, biotechnology research, and precision breeding for cassava.

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Section: Discussionmentioning

confidence: 99%

Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava

Tuo

Yao

et al. 2023

Plant Methods

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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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“…On infection with PlAMV‐GFP, GFP is expressed from sgRNA2 as a fusion protein (Figure 2b). An sgRNA duplication strategy, used for the development of other potexvirus vectors (Abrahamian et al, 2021), is difficult to apply to PlAMV due to its overlapping ORF4 and ORF5 (Minato et al, 2014).…”

Section: Genome Organization and Proteinsmentioning

confidence: 99%

“…An sgRNA duplication strategy, used for the development of other potexvirus vectors (Abrahamian et al, 2021), is difficult to apply to…”

Section: Genome Organization and Gene Expressionmentioning

confidence: 99%

Plantago asiatica mosaic virus: An emerging plant virus causing necrosis in lilies and a new model RNA virus for molecular research

Komatsu

2022

Molecular Plant Pathology

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Taxonomy: Plantago asiatica mosaic virus belongs to the genus Potexvirus in the family Alphaflexiviridae of the order Tymovirales. Virion and genome properties: Plantago asiatica mosaic virus (PlAMV) has flexuous virions of approximately 490-530 nm in length and 10-15 nm in width. The genome of PlAMV consists of a single-stranded, positive-sense RNA of approximately 6.13 kb. It contains five open reading frames (ORFs 1-5), encoding a putative viral polymerase (RdRp), movement proteins (triple gene block proteins, TGBp1-3), and coat protein (CP), respectively. Host range: PlAMV has an exceptionally wide host range and has been isolated from various wild plants, including Plantago asiatica, Nandina domestica, Rehmannia glutinosa, and other weed plants. Experimentally PlAMV can infect many plant species including Nicotiana benthamiana and Arabidopsis thaliana. It also infects ornamental lilies and frequently causes severe necrotic symptoms. However, host range varies depending on isolates, which show significant biological diversity within the species. Genome diversity: PlAMV can be separated into five clades based on phylogenetic analyses; nucleotide identities are significantly low between isolates in the different clades. Transmission: PlAMV is not reported to be transmitted by biological vectors. Virions of PlAMV are quite stable and it can be transmitted efficiently by mechanical contact. Disease symptoms: PlAMV causes red-rusted systemic necrosis in ornamental lilies, but it shows much weaker, if any, symptoms in wild plants such as P. asiatica. Control: Control of the disease caused by PlAMV is based mainly on rapid diagnosis and elimination of the infected bulbs or plants.

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Development and optimization of a pepino mosaic virus-based vector for rapid expression of heterologous proteins in plants

Cited by 5 publications

References 58 publications

Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava

Development of cassava common mosaic virus-based vector for protein expression and gene editing in cassava

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

Plantago asiatica mosaic virus: An emerging plant virus causing necrosis in lilies and a new model RNA virus for molecular research

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