Virus-induced gene silencing is an effective tool for assaying gene function in the basal eudicot species Papaver somniferum (opium poppy).
Virus-induced gene silencing (VIGS) is an attractive method for assaying gene function in species that are resistant to conventional genetic approaches. However, VIGS has been shown to be effective in only a few, closely related plant species. Tobacco rattle virus (TRV), a bipartite RNA virus, has a wide host range and so in principle could serve as an efficient vector for VIGS in a diverse array of plant species. Here we show that a vector based on TRV sequences is effective at silencing the endogenous phytoene desaturase (PapsPDS) gene in Papaver somniferum (opium poppy). We show that this vector does not compromise the growth or reproduction of poppy and the plants did not display viral symptoms. The silencing of PapsPDS resulted in a significant reduction in PapsPDS mRNA and a concomitant photobleached phenotype. The ability to rapidly assay gene function in P. somniferum will be valuable in manipulation of the opiate pathway in this pharmaceutically important species. We suggest that our vacuum infiltration method used to deliver TRV-based vectors into poppy is a promising approach for expanding VIGS to diverse angiosperm species in which traditional delivery methods fail to induce VIGS. Furthermore, these studies demonstrate the utility of TRV-VIGS for probing gene function in a basal eudicot species that is phylogenetically distant from model plant species.
Hileman
Lena C
LC
Department of Molecular, Cellular and Developmental Biology, PO Box 208104, Yale University, New Haven, CT 06520-8104, USA.
Drea
Sinéad
S
Martino
Gemma
G
Litt
Amy
A
Irish
Vivian F
VF
D002874:Chromosome Mapping; D032461:Chromosomes, Plant; D035264:Flowers / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D017343:Genes, Plant / Q000235:genetics* / Q000502:physiology*; D005822:Genetic Vectors / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010207:Papaver / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism* / Q000821:virology; D038761:Photobleaching; D010942:Plant Viruses / Q000235:genetics / Q000502:physiology*
D010088:Oxidoreductases; C022398:phytoene dehydrogenase