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Licciardello C.,Cra Centro Of Ricerca Per Lagrumicoltura E Le Colture Mediterranee | Russo M.P.,Cra Centro Of Ricerca Per Lagrumicoltura E Le Colture Mediterranee | Reforgiato Recupero G.,Cra Centro Of Ricerca Per Lagrumicoltura E Le Colture Mediterranee | Bernardi J.,Catholic University of Piacenza | And 4 more authors.
Acta Horticulturae | Year: 2011

A flesh-specific oligonucleotide custom array was designed to study gene expression during blood orange ripening. The array included 301 probes derived from a subtracted SSH library, a cDNA-AFLP collection, and a set of regulatory genes from the HarvEST Citrus database. The custom array was hybridized using fruits of 'Moro', a blood cultivar, and 'Cadenera', a common one, at three different ripening stages: the immature phase, the halfway point of maturation (corresponding to the start of 'Moro' pigmentation) and the full ripening. Among 301 probes, 27 in total, corresponding to 20 different transcripts, gave indication of differential expression in stage-stage and/or cultivar-cultivar comparisons. Transcripts encoding for anthocyanins biosynthesis represented most of the total over-expressed probes. The remaining differentially expressed transcripts were functionally associated with primary metabolism as flavour biosynthesis, defense and signal transduction. The expressed products indicating differentially expression were confirmed by Real time RT-PCR. The main novelty of the customized array here proposed is the use of expressed sequences specifically derived from blood orange flesh, integrating collections from other cultivars to study expression levels in fruits of different cultivars in ripening stages. This is of support to provide information about processes related to anthocyanin pigmentation in fruit flesh. To investigate the involvement of a member of the Glutathione S-Transferase gene family in the vacuolarization of anthocyanins, the SemiQ RT-PCR analysis was performed on various tissues (albedo, flavedo, flesh, young and adult leaves, ovary) of 'Moro' and Cadenera, comparing in silico predictions and molecular approaches. Real time RTPCR was conducted on different pigmented and not pigmented tissues of lemon and sweet oranges, showing the involvement fruit-specific (flesh and rind) of a GST in the anthocyanins pathway.

Frasca G.,University of Catania | Panico A.M.,University of Catania | Bonina F.,University of Catania | Messina R.,University of Catania | And 4 more authors.
Natural Product Research | Year: 2010

In the present study, a complex of compounds (red orange complex, ROC), obtained from three red orange varieties (Citrus sinensis varieties: Moro, Tarocco and Sanguinello), containing cyanidin glycosides, hydroxycinnamic acids, flavanone glycosides and ascorbic acid, was screened to discover new lead compounds in the suppression of the production of key molecules released during inflammatory events in interleukin-1 (IL-) stimulated human primary chondrocytes. The expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX)-2 and intercellular adhesion molecule-1 (ICAM-1), and the release of nitric oxide, prostaglandin (PG)E2 and interleukin-8 (IL-8) were determined. Indomethacin was used as an anti-inflammatory drug reference. ROC acts as a potent inhibitor of iNOS and COX-2 gene expression while also suppressing the production of PGE2 and nitrite in human chondrocytes. In addition, ROC induces a significant decrease in ICAM expression and IL-8 release. These findings suggest that ROC exerts anti-inflammatory effects probably through the suppression of COX-2 and iNOS expression. © 2010 Taylor & Francis.

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