iTRAQ-Based Quantitative Proteomics of Developing and Ripening Muscadine Grape Berry

Abstract

Grapes are among the widely cultivated fruit crops in the world. Grape berries like other nonclimacteric fruits undergo a complex set of dynamic, physical, physiological, and biochemical changes during ripening. Muscadine grapes are widely cultivated in the southern United States for fresh fruit and wine. To date, changes in the metabolites composition of muscadine grapes have been well documented; however, the molecular changes during berry development and ripening are not fully known. The aim of this study was to investigate changes in the berry proteome during ripening in muscadine grape cv. Noble. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS was used to detect statistically significant changes in the berry proteome. A total of 674 proteins were detected, and 76 were differentially expressed across four time points in muscadine berry. Proteins obtained were further analyzed to provide information about its potential functions during ripening. Several proteins involved in abiotic and biotic stimuli and sucrose and hexose metabolism were upregulated during berry ripening. Quantitative real-time PCR analysis validated the protein expression results for nine proteins. Identification of vicilin-like antimicrobial peptides indicates additional disease tolerance proteins are present in muscadines for berry protection during ripening. The results provide new information for characterization and understanding muscadine berry proteome and grape ripening.

Functional and Integrative Analysis of the Proteomic Profile of Radish Root under Pb Exposure

“…Isobaric tags for relative and absolute quantification (iTRAQ) is a robust mass spectrometry of protein quantitative technology, which can perform relative and absolute quantification in up to eight samples in parallel (Bindschedler and Cramer, 2011; Glibert et al, 2014). Recently, iTRAQ has been widely used for large-scale quantitative plant proteomic studies in exploration of various metabolic processes at the post-transcriptional level (Kambiranda et al, 2013; Martínez-Esteso et al, 2013) in response to various stresses (Yang et al, 2014; Li et al, 2015; Fu et al, 2016). Additionally, iTRAQ-based proteomics has been proven as a powerful method for unraveling the molecular regulatory networks involved in interactions between heavy metals and plant species, and a set of HM-responsive candidate proteins have been successfully identified.…”

Functional analysis of differentially expressed proteins in Chinese bayberry ( Myrica rubra Sieb. et Zucc.) fruits during ripening

“…Phosphoglucomutase (PGM, spot 4), which catalyzes the interconversion of glucose 1-phosphate and glucose 6-phosphate, was significantly increased in abundance during fruit ripening. Enolase (ENO, spot 44) is a key enzyme for phosphoenolpryruvate (PEP) generation because it participates in the conversion of the cytosolic pool of 2-phosphoglycerate to PEP (Kambiranda, Katam, Basha, & Siebert, 2014). In agreement with this scenario, ENO level strongly increased during Chinese bayberry ripening.…”

CXCL4 Contributes to the Pathogenesis of Chronic Liver Allograft Dysfunction

“…Extracted proteins were assayed using the BCA method [13] and digested according to the FASP method [14]. iTRAQ labeling was performed according to the manufacturer's instructions (Applied Biosystems).…”