Research article Exogenous application of a lipid transfer protein-jasmonic acid complex induces protection of grapevine towards infection by Botrytis cinerea T. Girault a,1,2 , J. Franc ¸ois a,1 , H. Rogniaux b , S. Pascal a,3 , S. Delrot a,4 , P. Coutos-The ´venot a , E. Gome `s a, * a Laboratoire de Physiologie, Biochimie et Biologie Mole ´culaire Ve ´ge ´tales, UMR CNRS-Universite ´ de Poitiers 6161, 40 avenue du recteur Pineau, 86022 Poitiers, France b Institut National de la Recherche Agronomique, UR 1268 Biopolyme `res, Interactions, Assemblage, Rue de la Ge ´raudie `re, B.P 71627, F-44316 Nantes, France Received 27 April 2007 Available online 9 October 2007 Abstract Type I plant lipid transfer proteins (LTPs) are small, basic, cystein-rich proteins involved in plant defense mechanisms. Five type I LTPs isoforms, named VvLTP1, 2, 3, 4 and 5 (Vitis vinifera lipid transfer proteins 1e5) were purified to homogeneity from the culture media of 41B grapevine cell suspension. The full sequence of isoforms 1, 3, 4 and 5 could be determined from mass spectrometry measurements of the enzymatically hydrolyzed proteins and from available VvLTP sequences. Phylogenetic analysis revealed that these proteins form two sub- groups, one with isoforms 1 and 4, and the second one with isoforms 3 and 5. The ability of the three most abundant ones (VvLTP1, 4 and 3) to interact with jasmonic acid (JA) was tested by fluorometric studies, showing that VvLTP4 was the most efficient to interact with this oxylipin. Exogenous application of the VvLTP4-JA complex on grapevine plantlets induced a high level (80.3  10.05%) of tolerance towards Botrytis cinerea, as compared with control plants (18.65  12.13%); whereas plants treated with JA or VvLTP4 alone exhibited a lower protection level (31.04  9.72% and 45.52  7.51% of protection, respectively). The results are discussed in the context of grapevine defense mechanisms. Ó 2007 Elsevier Masson SAS. All rights reserved. Keywords: Botrytis cinerea; Grapevine; Jasmonic acid; Lipid transfer proteins; Plant-pathogen interactions 1. Introduction Plants possess both preformed and inducible defense mech- anisms. Among the latter, expression of pathogenesis-related (PR) proteins is one of the most common responses of plants, following its encounter with a pathogen [1,2]. Currently, the PR-proteins are grouped in 17 families, based on their biolog- ical and biochemical properties [1]. Some PR-protein families, such as the PR-17 act through a still unknown mechanism, whereas several other have been reported to limit pathogen growth and/or activity. For example, PR-2 (b-endoglucanases) and PR-3, -4, -8 and 11 (chitinases) act against pathogenic fungi by degrading their cell wall [3]. Defensins (PR-12) and thionins (PR-13) exhibit a broad spectrum of antibacterial and antifungal activities [4,5]. Non-specific lipid transfer proteins (nsLTPs) constitute the PR-14 family [2]. These cystein-rich proteins have a secretion signal peptide at their N-terminal part, which confers them an extracellular localization; they appear to be rather ubiquitous in the plant kingdom, and are encoded by fairly large multi- genic families (for reviews, see refs. [6,7]). In Arabidopsis, * Corresponding author. Tel.: þ33 5 4936 6398. E-mail address: eric.gomes@univ-poitiers.fr (E. Gome `s). 1 These authors contributed equally to this work. 2 Present address: Laboratoire de Morphoge ´ne `se des Ligneux, UMR INRA-INH-Universite ´ d’Angers SAGAH A462, 2 Boulevard Lavoisier, 49045 Angers, France. 3 Present address: Institut des Sciences de la Vigne et du Vin, UMR Ecophy- siologie et Ge ´nomique Fonctionnelle de la Vigne, INRA, Domaine de la Grande Ferrade, 33883 Villenave d’Ornon, France. 4 Present address: Laboratoire de Biogene `se Membranaire, UMR CNRS- Universite ´ Bordeaux II 5200, 146 rue Leo Saignat, F-33076 Bordeaux, France. 0981-9428/$ - see front matter Ó 2007 Elsevier Masson SAS. All rights reserved. doi:10.1016/j.plaphy.2007.10.005 Available online at www.sciencedirect.com Plant Physiology and Biochemistry 46 (2008) 140e149 www.elsevier.com/locate/plaphy