Proteomic analysis validates previous findings on wounding-responsive plant hormone signaling and primary metabolism contributing to the biosynthesis of secondary metabolites based on metabolomic analysis in harvested broccoli (Brassica oleracea L. var. italica).

The plant wound-response is a complex process that generates physiological modifications for protecting the wounded tissue. In this study, tandem mass tag (TMT)-based quantitative proteomic analysis was performed to clarify the comprehensive molecular mechanism for the wound-response of broccoli subjected to two wounding intensities (0.04 and 1.85 m2 kg-1 for florets and shreds, respectively). Furthermore, integrated proteomic and metabolomic analysis was performed to reveal the interaction among the critical metabolic pathway responses to wounding. The results show that a total of 399 proteins and 266 proteins were identified as differentially expressed proteins (DEPs) in florets and shreds broccoli compared to control, respectively. Furthermore, 167 DEPs were detected in shreds broccoli compared to the florets broccoli. Salicylic acid (SA) and ethylene (ET) biosynthesis were more susceptible to being induced by wounding with lower intensities, whereas, phenylpropanoid biosynthesis, aliphatic glucosinolate synthesis and jasmonic acid (JA) biosynthesis were more susceptible to being activated by wounding with higher intensities. The activation of starch and sucrose metabolism, TCA cycle, glycolysis, pentose phosphate could provide carbon sources and ATP for the production of amino acids including phenylalanine, valine, threonine, isoleucine, L-methionine, methionine and histidine. The motivation of carbohydrate metabolic pathways and amino acid biosynthesis-related pathways promotes the precursor levels for phenolic substances and glucosinolate synthesis. Furthermore, the accumulation of SA, ET and JA may activated secondary metabolite biosynthesis through the regulation of critical proteins involved in the corresponding metabolic pathways.