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The expression of tomato prosystemin gene in tobacco plants highly ...

The expression of tomato prosystemin gene in tobacco plants highly ...

The expression of tomato prosystemin gene in tobacco plants highly

The expression of tomato prosystemin gene in tobacco plants highly affects host proteomic repertoire Mariapina Rocco a,1 , Giandomenico Corrado b,1 , Simona Arena c , Chiara D'Ambrosio c , Claudia Tortiglione b,2 , Stefano Sellaroli d , Mauro Marra d, ⁎, Rosa Rao b, ⁎, Andrea Scaloni c, ⁎ a Department of Biological and Environmental Sciences, University of Sannio, Benevento, Italy b Department of Soil, Plant, Environment and Animal Production, University of Naples Federico II, Portici, Naples, Italy c Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, via Argine 1085, 80147 Naples, Italy d Department of Biology, University of Rome “Tor Vergata”, Rome, Italy ARTICLE INFO ABSTRACT Article history: Received 22 February 2008 Accepted 22 April 2008 Keywords: Systemin Gene expression Prohormone processing Transgenic plants 1. Introduction Plants protect themselves against insect herbivores attack by activating a set of defence genes, both in wounded and distal unwounded leaves [1]. Some of these genes encode for JOURNAL OF PROTEOMICS 71 (2008) 176– 185 available at www.sciencedirect.com www.elsevier.com/locate/jprot Systemin, an octadecapeptide isolated from tomato, is a primary signal molecule involved in the local and systemic responses to pest attack, elicited by activation of a set of defence genes. It derives from processing of prosystemin, a prohormone of almost 200 amino acids. Prosystemin orthologues have been found in other Solanaceae species but not in tobacco, where are present hydroxyproline-rich peptides functionally but not structurally related to tomato systemin. Molecular events leading to the release of signalling peptides from protein precursors are unknown in plants; the occurrence of a family of signal molecules suggests that initiation of wound response may involve different processing mechanisms. It has been previously shown that the protein product from an engineered tomato prosystemin gene is processed in tobacco, thus suggesting that the components responsible for its posttranslational modifications are present in this species. By analyzing analysing the proteome repertoire of transformed tobacco plant leaves with 2-DE, here we demonstrate that the constitutive expression of the tomato prosystemin gene highly affected host protein synthesis. In particular, engineered plants showed a number of differentially synthesized proteins that were identified by PMF MALDI-TOF and µLC-ESI-IT-MS/MS experiments as polypeptide species involved in protection from pathogens and oxidative stress, or in carbon/energy metabolism. Significant differences in over-produced proteins were observed with respect to previous data reported on systemin-engineered tomato plants. Our results strongly support the need of using proteomic approaches during systematic analysis of plant tissues to investigate the principle of substantial equivalence in transgenic plants expressing a transgene coding for a signalling molecule. © 2008 Elsevier B.V. All rights reserved. proteinase inhibitor (PI) and polyphenol oxidase (PPO) molecules that perturb the normal digestive processes of herbivores [2], while others are involved in the production of volatile compounds exerting attraction of natural enemies of herbivores and therefore involved in indirect defence ⁎ Corresponding authors. E-mail addresses: Mauro.Marra@uniroma2.it (M. Marra), rao@cds.unina.it (R. Rao), andrea.scaloni@ispaam.cnr.it (A. Scaloni). 1 These authors gave an equal contribution to this work. 2 Current address: Institute of Cybernetics “E. Caianiello”, National Research Council, 80078, Pozzuoli, Naples, Italy. 1874-3919/$ – see front matter © 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.jprot.2008.04.003

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