Abstract

Plant cyclic nucleotides (cGMP and cAMP) have been reported to be involved in several physiological processes, but the mechanisms of action and the following signal transduction events, are still poorly characterized. The main object of the 2010 FIRB project is the study of cyclic nucleotide roles in signalling events following biotic stress in Arabidopsis thaliana.For this aim transgenic plants with altered levels of both cGMP and cAMP, have been generated. Specifically, plants over-expressing a mammalian guanylate cyclase (GC) or a phosphodiesterase (PDE) to alter the cGMP levels, and plants over-expressing a chimeric protein namely "cAMP-sponge" (cAS), to buffer the cAMP, were produced. Moreover, in order to measure both in vivo and in vitro the cNMP levels and dynamics, different approaches have been followed: i) production of transgenic lines expressing genetically encoded FRET-based probes (EPACs and Red-cGES) and ii) set up of alpha screen technology for plant tissue. To gain insight into the inter-relationships of cNMPs and other second messengers in the plant signalling events, we then crossed the plants with altered levels of cNMPs with Arabidopsis transgenic lines expressing the genetically encoded probes Cameleon (for in vivo Ca2+ detection) and roGFP2 (for in vivo detection of redox potential). In Arabidopsis, it has been reported that Ca2+, redox status and cNMPs play important signalling roles in response to the avirulent attack of Pseudomonas syringae. Our preliminary results, using Cameleon and roGFP2 wild type plants, showed a transient increase of both Ca2+ and oxidation status upon avirulent attack of Pseudomonas syringae. Biochemical analyses confirmed that both Glutathione and Ascorbate pools were more oxidized upon such attack. In order to study if and how the cNMPs play a role in the regulation of Ca2+ and redox status similar experiments will be performed with the different generated lines with altered levels of cNMPs. Finally, to understand if the altered levels of cNMPs can affect gene expression in response to the considered pathogen attack, a transcriptomic and proteomic characterization of the transgenic lines will be performed. Following this multidisciplinary approach, the understanding of the specific biological responses regulated by cAMP and cGMP and the downstream signalling events actually represents a big challenge in plants

Autore Pugliese

• Blanco Emanuela

Tutti gli autori

• SABETTA W.; DEPINTO M.C.; VANDELLE E.; BELLIN D.; LOCATO V.; TISI A.; FINCATO P.; COSTA A.; BLANCO E.

Titolo volume/Rivista

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Anno di pubblicazione

2014

ISSN

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ISBN

978-88-904570-4-3

Numero di citazioni Wos

Nessuna citazione

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Settori ERC

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Codici ASJC

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