<i>Foxtail Mosaic Virus</i>-Induced Gene Silencing in Monocot Plants

Liu, Na; Xie, Kui; Jia, Qi; Zhao, Jinping; Chen, Tianyuan; Li, Huangai; Xiang, Wei; Diao, Xianmin; Hong, Yiguo; Liu, Yule

doi:10.1104/pp.16.00010

Cited by 93 publications

(104 citation statements)

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“…Previous work by us and others demonstrated that FoMV is capable of systemic infection and inducing VIGS in maize (Liu et al, ; Mei & Whitham, ; Mei et al, ) or expressing proteins (Bouton et al, ). FoMV is a member of the genus Potexvirus , which has a single‐stranded, positive‐sense genomic RNA.…”

Section: Introductionmentioning

confidence: 93%

“…Our original FoMV‐VIGS vector carried the insertion site for target gene fragments immediately after the stop codon of ORF5, which worked well for VIGS, but it cannot be used for gene expression (Mei et al, ). The FoMV‐VIGS vector reported by Liu et al () is designed to carry target sequences for silencing under the control of a duplicated FoMV CP subgenomic promoter. Inverted‐repeats carried at this position were most efficient at inducing VIGS (Liu et al, ).…”

Section: Introductionmentioning

confidence: 99%

“…The FoMV‐VIGS vector reported by Liu et al () is designed to carry target sequences for silencing under the control of a duplicated FoMV CP subgenomic promoter. Inverted‐repeats carried at this position were most efficient at inducing VIGS (Liu et al, ). Bouton et al () also duplicated the CP promoter and demonstrated that FoMV could be used to transiently express marker and fungal effector proteins from this position in N. benthamiana , wheat, and maize.…”

Section: Introductionmentioning

confidence: 99%

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Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Beernink

Ellison

et al. 2019

Plant Direct

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Plant viruses can be engineered to carry sequences that direct silencing of target host genes, expression of heterologous proteins, or editing of host genes. A set of foxtail mosaic virus (FoMV) vectors was developed that can be used for transient gene expression and single guide RNA delivery for Cas9‐mediated gene editing in maize, Setaria viridis, and Nicotiana benthamiana. This was accomplished by duplicating the FoMV capsid protein subgenomic promoter, abolishing the unnecessary open reading frame 5A, and inserting a cloning site containing unique restriction endonuclease cleavage sites immediately after the duplicated promoter. The modified FoMV vectors transiently expressed green fluorescent protein (GFP) and bialaphos resistance (BAR) protein in leaves of systemically infected maize seedlings. GFP was detected in epidermal and mesophyll cells by epifluorescence microscopy, and expression was confirmed by Western blot analyses. Plants infected with FoMV carrying the bar gene were temporarily protected from a glufosinate herbicide, and expression was confirmed using a rapid antibody‐based BAR strip test. Expression of these proteins was stabilized by nucleotide substitutions in the sequence of the duplicated promoter region. Single guide RNAs expressed from the duplicated promoter mediated edits in the N. benthamiana Phytoene desaturase gene, the S. viridis Carbonic anhydrase 2 gene, and the maize HKT1 gene encoding a potassium transporter. The efficiency of editing was enhanced in the presence of synergistic viruses and a viral silencing suppressor. This work expands the utility of FoMV for virus‐induced gene silencing (VIGS), virus‐mediated overexpression (VOX), and virus‐enabled gene editing (VEdGE) in monocots.

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Section: Introductionmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Beernink

Ellison

et al. 2019

Plant Direct

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“…Viruses adapted to alter gene expression in monocots lag behind the number available for dicot plants. Vectors harboring Brome mosaic virus and Barley stripe mosaic virus are used for VIGS (Ding et al, 2006;Scofield and Nelson, 2009;Yuan et al, 2011), and Foxtail mosaic virus was shown recently to be effective for VIGS in C4 grasses, including foxtail millet (Setaria italica), maize (Zea mays), and sorghum (Liu et al, 2016;Mei et al, 2016). The organization of these viruses may be more suited to VIF by silencing TFL1 homologs.…”

Section: Vif: a Powerful Tool In A Spectrum Of Recalcitrant Plants?mentioning

confidence: 99%

Virus-Induced Flowering: An Application of Reproductive Biology to Benefit Plant Research and Breeding

et al. 2016

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“…Such plant virus-based technology can be applied to facilitate or impede gene expression, resulting in gain-or loss-offunction phenotypes. Although virus-based technology was initially exploited for the purpose of high-level production of foreign proteins, such as recombinant subunit vaccines and pharmaceutical proteins for molecular pharming (Scholthof et al, 1996;Porta and Lomonossoff, 2002), plant RNA and DNA virus-based techniques such as small interfering RNA-mediated virus-induced posttranscriptional gene silencing (VIGS) has been extensively utilized to silence genes for functional genomic studies in dicots and monocots, including plants and crops recalcitrant to classical forward or reverse genetic manipulation (Lindbo et al, 1993;Kumagai et al, 1995;Ruiz et al, 1998;Liu et al, 2002Liu et al, , 2016Becker and Lange, 2010;Senthil-Kumar and Mysore, 2011;Qin et al, 2015). Various virus-based technologies have been developed such as VIGS, microRNA-based VIGS (Tang et al, 2010;Chen et al, 2015aChen et al, , 2015c, virus-based microRNA silencing (Sha et al, 2014), and virus-induced transcriptional gene silencing (Kanazawa et al, 2011;Chen et al, 2015b).…”

mentioning

confidence: 99%

A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes

Cheng

Chen²,

Shen³

et al. 2017

Plant Physiol.

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Virus-induced flowering (VIF) uses virus vectors to express Flowering Locus T (FT) to induce flowering in plants. This approach has recently attracted wide interest for its practical applications in accelerating breeding in crops and woody fruit trees. However, the insight into VIF and its potential as a powerful tool for dissecting florigenic proteins remained to be elucidated. Here, we describe the mechanism and further applications of Potato virus X (PVX)-based VIF in the short-day Nicotiana tabacum cultivar Maryland Mammoth. Ectopic delivery of Arabidopsis (Arabidopsis thaliana) AtFT by PVX/AtFT did not induce the expression of the endogenous FT ortholog NtFT4; however, it was sufficient to trigger flowering in Maryland Mammoth plants grown under noninductive long-day conditions. Infected tobacco plants developed no systemic symptoms, and the PVX-based VIF did not cause transgenerational flowering. We showed that the PVX-based VIF is a much more rapid method to examine the impacts of single amino acid mutations on AtFT for floral induction than making individual transgenic Arabidopsis lines for each mutation. We also used the PVX-based VIF to demonstrate that adding a His-or FLAG-tag to the N or C terminus of AtFT could affect its florigenic activity and that this system can be applied to assay the function of FT genes from heterologous species, including tomato (Solanum lycopersicum) SFT and rice (Oryza sativa) Hd3a. Thus, the PVX-based VIF represents a simple and efficient system to identify individual amino acids that are essential for FT-mediated floral induction and to test the ability of mono-and dicotyledonous FT genes and FT fusion proteins to induce flowering.Modified plant viruses have emerged as powerful tools for dissecting gene function in plants. Such plant virus-based technology can be applied to facilitate or impede gene expression, resulting in gain-or loss-offunction phenotypes. Although virus-based technology was initially exploited for the purpose of high-level production of foreign proteins, such as recombinant subunit vaccines and pharmaceutical proteins for molecular pharming (Scholthof et al., 1996;Porta and Lomonossoff, 2002), plant RNA and DNA virus-based techniques such as small interfering RNA-mediated virus-induced posttranscriptional gene silencing (VIGS) has been extensively utilized to silence genes for functional genomic studies in dicots and monocots, including plants and crops recalcitrant to classical forward or reverse genetic manipulation (Lindbo et al

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Foxtail Mosaic Virus-Induced Gene Silencing in Monocot Plants

Cited by 93 publications

References 53 publications

Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Protein expression and gene editing in monocots using foxtail mosaic virus vectors

Virus-Induced Flowering: An Application of Reproductive Biology to Benefit Plant Research and Breeding

A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes

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