|Riassunto in Inglese:|
Tomato (Solanum lycopersicum) is a worldwide crop and Italy is the major producer among the Mediterranean Basin countries, leading the European Union market.
A large number of viruses have been reported to affect tomato, most of them considered critical since they cause severe economic and production losses.
Therefore, the multiple detection and identification of viruses and their strains affecting tomato crops is of critical importance to plant quarantine and certification programs world-wide.
We have developed a multiplex genomic strategy, based on DNA microarray second generation chip using Combimatrix platform 40-mer oligonucleotide probes, for screening the main tomato-infecting viruses, focusing on viruses with the highest economic relevance in Italy (Tiberini et al., 2010).The aim of this new diagnostic tool has been to facilitate the simultaneous detection and genotyping of the viruses and their strains from infected plant tissue or biological samples, in order to reduce analyses needed for assessing the presence/absence of viruses.
A total of 539 probes were designed for simultaneous and multiple detection and differentiation of 10 different viruses and their strains: Impatiens necrotic spot virus (INSV) and Tomato spotted wilt virus (TSWV) (Family: Bunyaviridae, Genus: Tospovirus) , Cucumber mosaic virus (CMV) ( Family: Bromoviridae, Genus: Cucumovirus), Pepino mosaic virus (PepMV) (Family: Flexiviridae, Genus: Potexvirus), Potato virus Y (PVY) (Family: Potyviridae, Genus: Potyvirus), Tomato infectious chlorosis virus (TICV) and Tomato chlorosis Virus (ToCV) (Family: Closteroviridae, Genus: Crinivirus), Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) (Genus: Tobamovirus) and Tomato yellow leaf curl virus (TYLCV) (Family: Geminiviridae, Genus: Begomovirus).
The microarray–based detection method, developed in this study, proved to be effective in detecting the above mentioned viruses (Tiberini et al., 2010), further allowing the genotyping of some of them, confirming the versatility and easiness of this technology in multiplex and routine analysis.