Parco Scientifico e Tecnologico della Sicilia s.c.p.a

Catania, Italy

Parco Scientifico e Tecnologico della Sicilia s.c.p.a

Catania, Italy

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Licciardello G.,Parco Scientifico e Tecnologico della Sicilia S.c.p.a. | Devescovi G.,Bacteriology Group | Bella P.,University of Catania | De Gregorio C.,Parco Scientifico e Tecnologico della Sicilia S.c.p.a. | And 3 more authors.
Chemical Engineering Transactions | Year: 2014

We analysed the draft genome sequence of Pseudomonas mediterranea CFBP 5447 in order to identify firstly the central metabolic pathways that convert fatty acids or carbohydrate intermediates into mcl-PHA and secondly the genes involved in glycerol metabolism (glpF, glpK, glpD, glpR). Absence of the glpF gene, which codifies for the "glycerol uptake facilitator protein", was highlighted. In order to understand the expression of the pha gene cluster, we investigated the promoter activity of phaC1, phaC2, phaZ, phaD and phaI genes. When glycerol was present as the carbon source, PI was found to be the most active promoter. Expression analysis of the knock-out mutant of the phaD gene, which is a transcriptional regulator belonging to the TetR family, showed that PhaD acts as an activator of the phaI promoter which, in turn, triggers the transcription of the phaIF operon. The activation of PC1, which controls the phaC1ZC2D, by PhaD, was less efficient than PI. © 2014, AIDIC Servizi S.r.l.


Licciardello G.,Parco Scientifico e Tecnologico della Sicilia s.c.p.a | Bella P.,University of Catania | Catara V.,University of Catania
Journal of Plant Pathology | Year: 2011

Pseudomonas corrugata and P. mediterranea are two closely related bacterial species both causal agents of tomato pith necrosis. To screen tomato planting material reliably, a quantitative real-time PCR assay was developed, to detect and/or discriminate both bacterial pathogens in a single tube. So, two species-specific primer/probe sets were designed on the sequences of two DNA fragments amplified by a previously reported specific PCR protocol. TaqMan real-time PCR assays were developed for individual (simplex PCR) and simultaneous (duplex PCR) amplifications. The assays were performed with the SmartCycler TD II System (Cepheid) and the fluorescence from both FAM and Texas Red channels were recorded at the annealing step. Specificity was tested with an extended range of P. corrugata and P. mediterranea strains, with other Pseudomonas spp. And with a number of tomato bacterial pathogens. The detection limit was approximately 10 cells per reaction for both bacteria, and quantification was linear over a six-log range. The duplex real-time PCR assay was validated on tomato plants artificially inoculated by pricking the stem with a strain of each species either separately or together.

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