The impact of sodium nitroprusside and ozone in kiwifruit ripening physiology: a combined gene and protein expression profiling approach

Ann Bot. 2015 Sep;116(4):649-62. doi: 10.1093/aob/mcv107. Epub 2015 Jul 8.

Abstract

Background and aims: 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).

Key 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.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actinidia / drug effects*
  • Actinidia / growth & development
  • Fruit / drug effects
  • Fruit / growth & development
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant / drug effects
  • Nitroprusside / pharmacology*
  • Ozone / pharmacology*
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Proteome*

Substances

  • Plant Proteins
  • Proteome
  • Nitroprusside
  • Ozone