Characterization of tomato PR1 processing in immunity: Evaluating the role of P69 on generation of the CAPE1 peptide


Vilafranca i Tello, Miquel


Bacterial wilt caused by the soilborne bacteria Ralstonia solanacearum is a worldwide concern. It affects every year hundreds of plant species, among them some agriculturally significant crops, hence having in some countries a collateral impact at both economic and social levels. However, in some species the plant presents a defensive response being resistant to the bacteria. This is the case of the tomato (Solanum lycopersicum) variant Hawaii 7996 although the mechanism behind the response is unclear. The Bacterial plant diseases and plant cell death group in CRAG is working on understanding it focusing on the secreted proteome of the tomato, precisely, the role that could play the pathogenesisrelated (PR) protein PR-1. Despite the fact that PR-1 is proved to have an anti-microbial activity, its mechanism of action has always been a big unknown. Therefore, this TFG was focused on analyzing the role that the proteases P69D and P69B could play on the on the activation by processing of PR-1, particularly on the cleavage and release of the CAPE1 peptide, constitutive of the PR-1, which recent studies spotlighted its involvement on the plant immune signaling. To do so, four classes of tomato PR-1 vectors, transformed in A. tumefaciens AsE cells, were transiently expressed in Nicotiana benthamiana leaves, and the resultant proteins were extracted and treated with P69. Finally, to overcome the drawback that entailed the transient expression of the limited protein expression, the concentration of L-arabinose required to optimize the expression of PR-1 was tested, for further assays to heterologously express the recombinant proteins in E. coli TOP10 cells.



Carnicer Heras, Marc
Sánchez Coll, Núria


IQS SE - Undergraduate Program in Biotechnology