Virus Induced Gene Silencing of a DEFICIENS Ortholog in Nicotiana Benthamiana
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Traditionally, developmental studies in plant biology have suffered from the lack of a convenient means to study gene function in non-model plant species. Here we show that virus-induced gene silencing (VIGS) is an effective new tool to study the function of orthologs of floral homeotic genes such as DEFICIENS(DEF) in non-model systems. We used a tobacco rattle virus (TRV)-based VIGS approach to study the function of the Nicotiana benthamiana DEFortholog (NbDEF). Silencing of NbDEFin N. benthamianausing TRV-VIGS was similar to that of Antirrhinum defand Arabidopsis ap3mutants and caused transformation of petals into sepals and stamens into carpels. Molecular analysis of the NbDEF-silenced plants revealed a dramatic reduction of the levels of NbDEFmRNA and protein in flowers. NbDEFsilencing was specific and has no effect on the mRNA levels of NbTM6, the closest paralog of NbDEF. A dramatic reduction of the levels of N. benthamiana GLOBOSA(NbGLO) mRNA and protein was also observed in flowers of NbDEF-silenced plants, suggesting that cross-regulation of this GLO-like gene by NbDEF. Taken together, our results suggest that NbDEF is a functional homolog of Antirrhinum DEF. Our results are significant in that they show that TRV efficiently induces gene silencing in young and differentiating flowers and that VIGS is a promising new tool for analyses of developmental gene function in non-model organisms.
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