OPEN ACCESS ARTICLE

This Article Free via Open Access: OA OA Full Text Full Text (PDF) OA All Versions of this Article: 145/4/1241    most recent pp.107.105916v1 Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via ISI Web of Science (2) Citing Articles via Google Scholar Google Scholar Articles by Spitzer, B. Articles by Vainstein, A. Search for Related Content PubMed PubMed Citation Articles by Spitzer, B. Articles by Vainstein, A. Agricola Articles by Spitzer, B. Articles by Vainstein, A. Related Collections Vector Systems for Plant Research and Biotechnology

Reverse Genetics of Floral Scent: Application of Tobacco Rattle Virus-Based Gene Silencing in Petunia^1,[OA]

Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture (B.S., M.M.B.Z., M.O., E.M., O.B., O.E., I.M., T.M., A.V.) and Department of Entomology, Faculty of Agricultural, Food and Environmental Quality Sciences (M.A., S.M.), Hebrew University of Jerusalem, Rehovot 76100, Israel; and Weizmann Institute of Science, Rehovot 76100, Israel (I.R., A.A.)

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 (Petunia hybrida), 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-based VIGS system to studies of floral scent. Suppression of the anthocyanin pathway via chalcone synthase silencing was used as a reporter, allowing easy visual identification of anthocyaninless silenced flowers/tissues with no effect on the level of volatile emissions. Use of tobacco rattle virus constructs containing target genes involved in phenylpropanoid volatile production, fused to the chalcone synthase 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, phenylacetaldehyde synthase, and the myb transcription factor ODORANT1. Because 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.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Alexander Vainstein (vain{at}agri.huji.ac.il).

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.105916

^* Corresponding author; e-mail vain{at}agri.huji.ac.il.

Received July 19, 2007; accepted August 16, 2007; published August 24, 2007.