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New Phytol

Title:		Optimized virus-induced gene silencing in Solanum nigrum reveals the defensive function of leucine aminopeptidase against herbivores and the shortcomings of empty vector controls
Author(s):		Hartl M; Merker H; Schmidt DD; Baldwin IT;
Address:		"Max Planck Institute for Chemical Ecology, Department of Molecular Ecology, Hans-Knoll-Strasse 8, D-07745 Jena, Germany"
Journal Title:		New Phytol
Year:		2008
Volume:		179
Issue:		2
Page Number:		356 - 365
DOI:		10.1111/j.1469-8137.2008.02479.x
ISSN/ISBN:		0028-646X (Print) 0028-646X (Linking)
Abstract:		"Virus-induced gene silencing (VIGS) enables high-throughput analysis of gene function in plants but is not universally applicable and requires optimization for each species. Here a VIGS system is described for Solanum nigrum, a wild relative of tomato and potato and a valuable model species for ecogenomics. The efficiency of the two most widely used Tobacco rattle virus (TRV) vectors to silence phytoene desaturase (PDS) in S. nigrum was tested. Additionally, the infiltration method and growth temperatures for gene silencing were optimized and the suitability of different control vectors evaluated. Using leucine aminopeptidase (LAP), a herbivore-induced protein, silencing efficiency and the applicability of silenced plants for herbivore feeding assays were assessed. Vacuum infiltration of seedlings with Agrobacterium carrying the vector, pYL156, proved the most efficient means of silencing genes. Empty-vector controls decreased plant growth but control vectors carrying a piece of noncoding sequence did not. Silencing LAP significantly increased the larval mass of Manduca sexta that fed on silenced plants. This VIGS protocol proved highly successful for S. nigrum, which should include control vectors carrying noncoding sequence as control treatments. Silencing LAP provided the first experimental evidence that LAP has a defensive function against herbivores"
Keywords:		"Animals Feeding Behavior Gene Expression Regulation, Plant Gene Silencing Genotype Larva Leucyl Aminopeptidase/*metabolism Manduca/physiology Plant Leaves Plant Proteins/genetics/metabolism Plant Viruses/pathogenicity Solanum nigrum/*metabolism Temperatur;"
Notes:		"MedlineHartl, Markus Merker, Holger Schmidt, Dominik D Baldwin, Ian T eng Research Support, Non-U.S. Gov't England 2008/12/17 New Phytol. 2008 Jul; 179(2):356-365. doi: 10.1111/j.1469-8137.2008.02479.x"