Titolo Progetto

"Dissecting the phloem response to a phloem-limited pathogen in grapevine" CUP: G53D23004200006 CODICE PROGETTO: 022MCEJP2

Programma di finanziamento

Fondo per il Programma Nazionale di Ricerca e Progetti di Rilevante Interesse Nazionale (PRIN)
D.D. 104 del 02/02/2022
PNRR M4.C2.1.1 Finanziato dall’Unione Europea Next Generation EU

Abstract

Grapevine Yellows (GY) are associated with phloem-restricted phytoplasmas, whose management relies exclusively on the preventive control of the vectors using compulsory insecticide application. Natural resistance resources to phytoplasma diseases are not known in grapevine, as well in other plants. An interesting aspect of the phytoplasma-plant interaction is recovery that is a spontaneous remission of symptoms in previously symptomatic plants. Genes modulated in recovered plants can be regarded as promising players in phytoplasma resistance, but the molecular mechanisms underlying recovery are still unknown. Despite transcriptome and proteome profiling have been previously applied to the study of plant response to phytoplasma, the pivotal molecular players during the interaction have not been identified yet, being masked by the large, complex transcript and protein profiles obtained by conventional bulk profiling. Thus, the aim of our project is to study the molecular mechanisms of plant-phytoplasma interaction focusing on the phloem. To reveal transcriptomic changes specifically at the infection site, our research group has recently carried out a combination of Laser Microdissection Pressure Catapulting and transcriptome profiling by RNA-seq and obtained a sub-set of genes that are differentially regulated in the grapevine phloem either upon infection by ‘Candidatus Phytoplasma solani’ or upon recovery. This will serve as foundation for the development of computational models of gene co-expression networks which could evidence crucial molecular players in the interaction that could then be verified by classical reverse genetics approaches. Transcript profile will be also mined to identify the presence of a pathogen effector-precursor or effector; thus follow-up analyses will be focused on both best plant candidate(s) and possible pathogen effectors or effector-precursors. Proteomics analysis of the phloem exudates and the leaf midrib is planned, thus proteomics coupled with phloem transcriptome profiling will allow to select the best candidate(s). In order to this information to be maximized and utilized to produce new relevant biological knowledge, our work plan includes follow-up analyses aimed at better defining the function of the selected gene(s) in a model pathosystem, ‘Ca. P. solani’-infected Solanum lycopersicum cv. Micro-Tom. A platform of CRISPR/CAS9-based transformation will be set up for silencing or overexpressing candidate genes in tomato. Transgenic plants constitutively expressing an effector gene, if identified, will be produced and phenotype analyzed. Focusing on the infection site represents the strength of our proposal. Project outcomes have the potential to improve significantly basic knowledge on general and specific plant defense mechanisms during phytoplasma infection, leading in the long term, to the development of more sustainable GY control strategies with evident beneficial effects on environment and public health.

Partenariato

• Università degli Studi di Udine
• Università degli Studi di Padova (Principal Investigator)

Importo del progetto

Importo totale del progetto        Euro 222.408,00

Importo del progetto Uniud        Euro 103.410,00

Finanziamento Uniud                Euro 28.193,00

Durata

• Data avvio progetto: 12/10/2023
• Data conclusione progetto: 11/10/2025