基于iTRAQ技术的采后乙烯利和1-甲基环丙烯处理对茭白线粒体蛋白质表达谱的影响  

Effect of postharvest ethylene and 1-methylcyclopropene treatments on protein profile in Zizania latifolia mitochondria based on iTRAQ labeling technique

作　　者：江凯[1] 罗海波 姜丽[3] 周涛[2] 陶明煊[2] 王韦华[3] 郁志芳[3] JIANG Kai;LUO Hai-bo;JIANG Li;ZHOU Tao;TAO Ming-xuan;WANG Wei-hua;YU Zhi-fang(Faculty of Food Science, Zhejiang Pharmaceutical College, Ningbo 315100, China;Jinling College, Nanjing Normal University, Nanjing 210097, China;College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, China)

机构地区：[1]浙江医药高等专科学校食品学院,浙江宁波315100 [2]南京师范大学金陵女子学院,江苏南京210097 [3]南京农业大学食品科学技术学院,江苏南京210095

出　　处：《江苏农业学报》2019年第1期173-181,共9页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金青年科学基金项目(31401612)

摘　　要：利用同位素标记相对和绝对定量(i TRAQ)标记结合二维液相色谱-串联质谱(2D-LC-MS/MS)技术,探索乙烯利(ETH)和1-甲基环丙烯(1-MCP)处理对茭白采后贮藏期间线粒体蛋白质表达谱的影响。共鉴定到1 908个特征肽段≥2的可信蛋白质,与贮藏0 d相比,对照、ETH和1-MCP处理的茭白贮藏3 d和6 d共有315个蛋白质具有2. 0倍以上显著差异(P<0. 05)。基因本体(GO)分子功能分析和KEGG通路分析结果显示,茭白采后贮藏3d,代谢途径、次生代谢产物生物合成、氨基酸生物合成、抗菌物质生物合成、碳代谢、光合作用中的碳固定、2-酮酸代谢、精氨酸生物合成、α-亚麻酸代谢、二羧酸代谢相关蛋白质显著富集;贮藏6 d,光合作用中的碳固定、α-亚麻酸代谢和二羧酸代谢途径减弱,丙氨酸-天冬氨酸-谷氨酸代谢、缬氨酸-亮氨酸-异亮氨酸降解和磷酸戊糖途径相关蛋白质显著富集。ETH处理后贮藏3 d,氧化磷酸化以及甘氨酸-丝氨酸-苏氨酸代谢相关蛋白质显著富集;贮藏6 d,丙氨酸-天冬氨酸-谷氨酸代谢、磷酸戊糖途径以及苯丙氨酸-酪氨酸-色氨酸生物合成代谢途径相关蛋白质显著富集。1-MCP处理后贮藏3 d,三羧酸循环、丙氨酸-天冬氨酸-谷氨酸代谢、缬氨酸-亮氨酸-异亮氨酸降解途径相关蛋白质显著富集;贮藏6 d,磷酸戊糖途径、C5支链二元酸代谢、缬氨酸-亮氨酸-异亮氨酸降解、丙氨酸-天冬氨酸-谷氨酸代谢途径相关蛋白质显著富集。以上结果表明,代谢途径、次生代谢产物生物合成、氨基酸生物合成、抗菌物质生物合成、碳代谢、光合作用中的碳固定、2-酮酸代谢、精氨酸生物合成、α-亚麻酸代谢和二羧酸代谢途径等可能与茭白采后衰老有关,三羧酸循环、氧化磷酸化、磷酸戊糖途径、C5支链酸代谢及相关氨基酸代谢途径可能在茭白采后衰老中发挥重要作用。Using isobaric tags for relative and absolute quantification( iT RAQ) labeling combined with2 D-LC-MS/MS techniques,the effects of ethylene( ETH)and 1-methylcyclopropene( 1-MCP) treatments on protein profile in Zizania latifolia mitochondria were explored. The results showed that a total of 1 908 proteins with two or more peptides were identified. Compared with the Z. latifolia stored for 0 d,there were 315 proteins in the Z. latifolia stored for 3 d and 6 d under control,ETH and 1-MCP treatments showed significant( P<0.05) change in relative quantitation based on two-fold difference. The gene ontology( GO) and kyoto encyclopedia of genes and genomes( KEGG) pathway analysis results of these differentially expressed proteins showed that the proteins associated with metabolic pathways,biosynthesis of secondary metabolites,biosynthesis of amino acids,biosynthesis of antibiotics,carbon metabolism,carbon fixation in photosynthesis,2-keto acid metabolism,arginine biosynthesis,alpha-linolenic acid metabolism and dicarboxylate metabolism in postharvest Z. latifolia mitochondria were significantly enriched during storage at 25 ℃ for three days. After six days of storage,carbon fixation in photosynthesis,alpha-linolenic acid metabolism and dicarboxylate metabolism pathways were weakened,while the proteins associated with valine-leucine-isoleucine degradation,alanine-aspartate-glutamate metabolism and pentose phosphate pathway( PPP) were significantly enriched. ETH treatment significantly enriched the proteins associated with the oxidative phosphorylation( OXPHOS),glycine-serine-threonine metabolism,alanine-aspartate-glutamate metabolism,PPP and phenylalanine-tyrosine-tryptophan biosynthesis pathways after three and six days of storage. MCP treatment significantly enriched proteins associated with the tricarboxylic acid( TCA) cycle,alanine-aspartate-glutamate metabolism,valine-leucine-isoleucine degradation,PPP,C5-branched dibasic acid metabolism pathways after three and six days of storage. These results indicated that t

关 键 词：茭白 线粒体 同位素标记相对和绝对定量(iTRAQ) 生物信息学 衰老 

分 类 号：S645.2[农业科学—蔬菜学]