Highly Efficient Virus-induced Gene Silencing (VIGS) in California Poppy (Eschscholzia californica): An Evaluation of VIGS as a Strategy to Obtain Functional Data from Non-model Plants

Wege, Stefanie; Scholz, Andrea; Gleissberg, Stefan; Becker, Annette

doi:10.1093/aob/mcm118

Cited by 113 publications

(89 citation statements)

References 28 publications

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“…We used the bipartite Tobacco rattle virus, in which RNA1 (or TRV1) encodes the viral replicase and the RNA-dependent RNA polymerase and RNA2 (or TRV2) encodes the coat protein and has a multiple cloning site for insertion of the endogenous target sequence. This approach has been successfully implemented in poppies (Hileman et al, 2005;Drea et al, 2007;Wege et al, 2007;Yellina et al, 2010) and other Ranunculales species Kramer et al, 2007;Di Stilio et al, 2010). In order to specifically silence each FUL-like paralog in opium poppy, we generated TRV2 constructs carrying a short fragment of PapsFL1 or PapsFL2 (Supplemental Fig.…”

Section: Silencing Of Papsful-like Genes Using Trv-vigsmentioning

confidence: 99%

“…Both taxa belong to the Papaveraceae, an early-diverging family of basal eudicots; therefore, they are evolutionary intermediates between the distantly related monocots and core eudicots. Importantly, both taxa are amenable to functional analysis using virus-induced gene silencing (VIGS; Hileman et al, 2005;Drea et al, 2007;Wege et al, 2007;Orashakova et al, 2009). Differences in floral morphology, in addition to the close relationship of these two species, make a comparison between them a robust platform from which to assess the role of FUL-like genes in basal eudicots.…”

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confidence: 99%

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PoppyAPETALA1/FRUITFULLOrthologs Control Flowering Time, Branching, Perianth Identity, and Fruit Development

Pabón‐Mora

Ambrose²,

Litt³

2012

Plant Physiology

108

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Several MADS box gene lineages involved in flower development have undergone duplications that correlate with the diversification of large groups of flowering plants. In the APETALA1 gene lineage, a major duplication coincides with the origin of the core eudicots, resulting in the euFUL and the euAP1 clades. Arabidopsis FRUITFULL (FUL) and APETALA1 (AP1) function redundantly in specifying floral meristem identity but function independently in sepal and petal identity (AP1) and in proper fruit development and determinacy (FUL). Many of these functions are largely conserved in other core eudicot euAP1 and euFUL genes, but notably, the role of APETALA1 as an "A-function" (sepal and petal identity) gene is thought to be Brassicaceae specific. Understanding how functional divergence of the core eudicot duplicates occurred requires a careful examination of the function of preduplication (FUL-like) genes. Using virus-induced gene silencing, we show that FUL-like genes in opium poppy (Papaver somniferum) and California poppy (Eschscholzia californica) function in axillary meristem growth and in floral meristem and sepal identity and that they also play a key role in fruit development. Interestingly, in opium poppy, these genes also control flowering time and petal identity, suggesting that AP1/FUL homologs might have been independently recruited in petal identity. Because the FUL-like gene functional repertoire encompasses all roles previously described for the core eudicot euAP1 and euFUL genes, we postulate subfunctionalization as the functional outcome after the major AP1/FUL gene lineage duplication event.

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Section: Silencing Of Papsful-like Genes Using Trv-vigsmentioning

confidence: 99%

mentioning

confidence: 99%

PoppyAPETALA1/FRUITFULLOrthologs Control Flowering Time, Branching, Perianth Identity, and Fruit Development

Pabón‐Mora

Ambrose²,

Litt³

2012

Plant Physiology

108

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“…SEI was digested with BamHI and XhoI and cloned into the equivalently digested pTRV2 vector to create pTRV2-SEI. Agrobacterium tumefaciens GV3101 was used to inoculate 3-week-old California poppy seedlings as described previously (Orashakova et al, 2009), and plants were grown under the conditions described earlier (Wege et al, 2007).…”

Section: Vigsmentioning

confidence: 99%

TheseirenaB Class Floral Homeotic Mutant of California Poppy (Eschscholzia californica) Reveals a Function of the Enigmatic PI Motif in the Formation of Specific Multimeric MADS Domain Protein Complexes

et al. 2013

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The products of B class floral homeotic genes specify petal and stamen identity, and loss of B function results in homeotic conversions of petals into sepals and stamens into carpels. Here, we describe the molecular characterization of seirena-1 (sei-1), a mutant from the basal eudicot California poppy (Eschscholzia californica) that shows homeotic changes characteristic of floral homeotic B class mutants. SEI has been previously described as EScaGLO, one of four B class-related MADS box genes in California poppy. The C terminus of SEI, including the highly conserved PI motif, is truncated in sei-1 proteins. Nevertheless, like the wild-type SEI protein, the sei-1 mutant protein is able to bind CArG-boxes and can form homodimers, heterodimers, and several higher order complexes with other MADS domain proteins. However, unlike the wild type, the mutant protein is not able to mediate higher order complexes consisting of specific B, C, and putative E class related proteins likely involved in specifying stamen identity. Within the PI motif, five highly conserved N-terminal amino acids are specifically required for this interaction. Several families lack this short conserved sequence, including the Brassicaceae, and we propose an evolutionary scenario to explain these functional differences.

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“…VIGS relies on transient as opposed to stable plant transformation; the spread of the silencing phenotype is governed by the speed of transmission of the silencing signal to new organs or within organs and, consequently, silenced tissues can be obtained rapidly. VIGS has been shown to work in a broad spectrum of recipient plant species, and technical procedures are relatively simple and easily adapted to high-throughput strategies; consequently, VIGS has become a popular reverse genetics tool in both model species and crops (Burch-Smith et al, 2006;Wang et al, 2006Wang et al, , 2007Nelson et al, 2007;Wege et al, 2007;Zhang et al, 2009). …”

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confidence: 99%

A Visual Reporter System for Virus-Induced Gene Silencing in Tomato Fruit Based on Anthocyanin Accumulation

et al. 2009

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Innes Centre, Colney, Norwich NR4 7UH, United Kingdom (E.B., C.M.)Virus-induced gene silencing (VIGS) is a powerful tool for reverse genetics in tomato (Solanum lycopersicum). However, the irregular distribution of the effects of VIGS hampers the identification and quantification of nonvisual phenotypes. To overcome this limitation, a visually traceable VIGS system was developed for fruit, comprising two elements: (1) a transgenic tomato line (Del/Ros1) expressing Antirrhinum majus Delila and Rosea1 transcription factors under the control of the fruitspecific E8 promoter, showing a purple-fruited, anthocyanin-rich phenotype; and (2) a modified tobacco rattle virus VIGS vector incorporating partial Rosea1 and Delila sequences, which was shown to restore the red-fruited phenotype upon agroinjection in Del/Ros1 plants. Dissection of silenced areas for subsequent chemometric analysis successfully identified the relevant metabolites underlying gene function for three tomato genes, phytoene desaturase, TomloxC, and SlODO1, used for proof of concept. The C-6 aldehydes derived from lipid 13-hydroperoxidation were found to be the volatile compounds most severely affected by TomloxC silencing, whereas geranial and 6-methyl-5-hepten-2-one were identified as the volatiles most severely reduced by phytoene desaturase silencing in ripening fruit. In a third example, silencing of SlODO1, a tomato homolog of the ODORANT1 gene encoding a myb transcription factor, which regulates benzenoid metabolism in petunia (Petunia hybrida) flowers, resulted in a sharp accumulation of benzaldehyde in tomato fruit. Together, these results indicate that fruit VIGS, enhanced by anthocyanin monitoring, can be a powerful tool for reverse genetics in the study of the metabolic networks operating during fruit ripening.

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Highly Efficient Virus-induced Gene Silencing (VIGS) in California Poppy (Eschscholzia californica): An Evaluation of VIGS as a Strategy to Obtain Functional Data from Non-model Plants

Cited by 113 publications

References 28 publications

PoppyAPETALA1/FRUITFULLOrthologs Control Flowering Time, Branching, Perianth Identity, and Fruit Development

PoppyAPETALA1/FRUITFULLOrthologs Control Flowering Time, Branching, Perianth Identity, and Fruit Development

TheseirenaB Class Floral Homeotic Mutant of California Poppy (Eschscholzia californica) Reveals a Function of the Enigmatic PI Motif in the Formation of Specific Multimeric MADS Domain Protein Complexes

A Visual Reporter System for Virus-Induced Gene Silencing in Tomato Fruit Based on Anthocyanin Accumulation

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