Tomato Prosystemin Is Much More than a Simple Systemin Precursor

SIMPLE SUMMARY: Prosystemin is a 200 amino acid precursor that releases, upon wounding and biotic attacks, an 18 amino acid peptide called Systemin. This peptide was traditionally considered as the principal actor of the resistance of tomato plants induced by triggering multiple defense pathways in response to a wide range of biotic/abiotic stress agents. Recent findings from our group discovered the disordered structure of Prosystemin that promotes the binding of different molecular partners and the possible activation of multiple stress-related pathways. All of our recent findings suggest that Prosystemin could be more than a simple precursor of Systemin peptide. Indeed, we hypothesized that it contains other sequences able to activate multiple stress-related responses. To verify this hypothesis, we produced a truncated Prosystemin protein deprived of the Systemin peptide and the relative deleted gene. Experiments with transgenic tomato plants overexpressing the truncated Prosystemin and with plants exogenously treated with the recombinant truncated protein demonstrated that both transgenic and treated plants modulated the expression of defense-related genes and were protected against a noctuid moth and a fungal pathogen. Taken together, our results demonstrated that Prosystemin is not a mere scaffold of Systemin, but itself contains other biologically active regions. ABSTRACT: Systemin (Sys) is an octadecapeptide, which upon wounding, is released from the carboxy terminus of its precursor, Prosystemin (ProSys), to promote plant defenses. Recent findings on the disordered structure of ProSys prompted us to investigate a putative biological role of the whole precursor deprived of the Sys peptide. We produced transgenic tomato plants expressing a truncated ProSys gene in which the exon coding for Sys was removed and compared their defense response with that induced by the exogenous application of the recombinant truncated ProSys (ProSys((1-178),) the Prosystemin sequence devoid of Sys region). By combining protein structure analyses, transcriptomic analysis, gene expression profiling and bioassays with different pests, we demonstrate that truncated ProSys promotes defense barriers in tomato plants through a hormone-independent defense pathway, likely associated with the production of oligogalacturonides (OGs). Both transgenic and plants treated with the recombinant protein showed the modulation of the expression of genes linked with defense responses and resulted in protection against the lepidopteran pest Spodoptera littoralis and the fungus Botrytis cinerea. Our results suggest that the overall function of the wild-type ProSys is more complex than previously shown, as it might activate at least two tomato defense pathways: the well-known Sys-dependent pathway connected with the induction of jasmonic acid biosynthesis and the successive activation of a set of defense-related genes, and the ProSys((1-178))-dependent pathway associated with OGs production leading to the OGs mediate plant immunity.