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Published on August 24, 2007; 10.1104/pp.107.105916

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Received July 19, 2007
Accepted August 16, 2007

Reverse Genetics of Floral Scent: Application of TRV-based Gene Silencing in Petunia

Ben Spitzer , Michal Moyal Ben Zvi , Marianna Ovadis , Elena Marhevka , Oren Barkai , Orit Edelbaum , Ira Marton , Tania Masci , Michal Alon , Shai Morin , Ilana Rogachev , Asaph Aharoni , and Alexander Vainstein *

The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, P.O. Box 12, Rehovot 76100, Israel; Department of Entomology, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, Rehovot 76100, Israel; Weizmann Institute of Science, P.O. Box 26, Rehovot 76100, Israel

* Corresponding author; email: vain{at}agri.huji.ac.il.

Floral fragrance is responsible for attracting pollinators as well as repelling pathogens and pests. As such, it is of immense biological importance. Molecular dissection of the mechanisms underlying scent production would benefit from the use of model plant systems with big floral organs that generate an array of volatiles and that are amenable to methods of forward and reverse genetics. One candidate is petunia, which has emerged as a convenient model system, and both RNAi and overexpression approaches, using transgenes, have been harnessed for the study of floral volatiles. Virus-induced gene silencing (VIGS) is characterized by a simple inoculation procedure and rapid results, relative to transgenesis. Here, we demonstrate the applicability of the Tobacco rattle virus (TRV)-based VIGS system to studies of floral scent. Suppression of the anthocyanin pathway via chalcone synthase (CHS) silencing was used as a reporter, allowing easy visual identification of "anthocyanin-less" silenced flowers/tissues with no effect on the level of volatile emissions. Use of TRV constructs containing target genes involved in phenylpropanoid volatile production, fused to the CHS reporter, allowed simple identification of flowers with suppressed activity of the target genes. The applicability of VIGS was exemplified with genes encoding S-adenosyl-L-methionine:benzoic acid/salicylic acid carboxyl methyltransferase (BSMT), phenylacetaldehyde synthase (PAAS) and the myb transcription factor ODORANT1. Since this high-throughput reverse-genetics approach was applicable to both structural and regulatory genes responsible for volatile production, it is expected to be highly instrumental for large-scale scanning and functional characterization of novel scent genes.




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