title: The impact of sodium nitroprusside and ozone in kiwifruit ripening physiology: a combined gene and protein expression profiling approach.
abstract: OBJECTIVE
  Despite their importance in many aspects of plant physiology, information about the function of oxidative and, particularly, of nitrosative signalling in fruit biology is limited. This study examined the possible implications of O3 and sodium nitroprusside (SNP) in kiwifruit ripening, and their interacting effects. It also aimed to investigate changes in the kiwifruit proteome in response to SNP and O3 treatments, together with selected transcript analysis, as a way to enhance our understanding of the fruit ripening syndrome.
  
  
  METHODS
  Kiwifruits following harvest were pre-treated with 100 μm SNP, then cold-stored (0 °C, relative humidity 95 %) for either 2 or 6 months in the absence or in the presence of O3 (0·3 μL L(-1)), and subsequently were allowed to ripen at 20 °C. The ripening behaviour of fruit was characterized using several approaches: together with ethylene production, several genes, enzymes and metabolites involved in ethylene biosynthesis were analysed. Kiwifruit proteins were identified using 2-D electrophoresis coupled with nanoliquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Expression patterns of kiwifruit ripening-related genes were also analysed using real-time quantitative reverse transcription-PCR (RT-qPCR).
  
  
  RESULTS
  O3 treatment markedly delayed fruit softening and depressed the ethylene biosynthetic mechanism. Although SNP alone was relatively ineffective in regulating ripening, SNP treatment prior to O3 exposure attenuated the O3-induced ripening inhibition. Proteomic analysis revealed a considerable overlap between proteins affected by both SNP and O3. Consistent with this, the temporal dynamics in the expression of selected kiwifruit ripening-related genes were noticeably different between individual O3 and combined SNP and O3 treatments.
  
  
  CONCLUSIONS
  This study demonstrates that O3-induced ripening inhibition could be reversed by SNP and provides insights into the interaction between oxidative and nitrosative signalling in climacteric fruit ripening.
authors.affiliation: School of Agriculture, Aristotle University of Thessaloniki, University Campus, 54124 Thessaloniki, Greece.
authors.forename: Georgia
authors.initials: G
authors.lastname: Tanou
authors.affiliation: School of Agriculture, Aristotle University of Thessaloniki, University Campus, 54124 Thessaloniki, Greece.
authors.forename: Ioannis S
authors.initials: IS
authors.lastname: Minas
authors.affiliation: School of Agriculture, Aristotle University of Thessaloniki, University Campus, 54124 Thessaloniki, Greece.
authors.forename: Evangelos
authors.initials: E
authors.lastname: Karagiannis
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, 41221 Larissa, Greece and.
authors.forename: Daniela
authors.initials: D
authors.lastname: Tsikou
authors.affiliation: CRCM, INSERM U1068, Institute Paoli-Calmettes, Aix-Marseille University, UM105, CNRS, UMR7258, 163 Luminy Av.F-13009 Marseille, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Audebert
authors.affiliation: Department of Biochemistry and Biotechnology, University of Thessaly, 41221 Larissa, Greece and.
authors.forename: Kalliope K
authors.initials: KK
authors.lastname: Papadopoulou
authors.affiliation: School of Agriculture, Aristotle University of Thessaloniki, University Campus, 54124 Thessaloniki, Greece, amolasio@agro.auth.gr.
authors.forename: Athanassios
authors.initials: A
authors.lastname: Molassiotis
mesh_terms: D029042:Actinidia / Q000187:drug effects* / Q000254:growth & development; D005638:Fruit / Q000187:drug effects / Q000254:growth & development; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D009599:Nitroprusside / Q000494:pharmacology*; D010126:Ozone / Q000494:pharmacology*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D020543:Proteome
chemical_list: D010940:Plant Proteins; D020543:Proteome; D009599:Nitroprusside; D010126:Ozone
keywords: Actinidia deliciosa; RNS; ROS; ethylene; fruit softening; gene expression; kiwifruit; mass spectrometry; nitrosative signalling; oxidative signalling; ozone; post-harvest; proteomics; reactive nitrogen species; reactive oxygen species; ripening; sodium nitroprusside