摘要
应用同位素标记相对和绝对定量(i TRAQ)技术结合液相色谱-串联质谱法(LC-MS/MS),分析桑树硬枝扦插生根不同阶段插穗基部皮层的蛋白质组差异,为从分子水平研究桑树扦插生根调控机制提供基础信息。在扦插初期、膨大期、发根期3个阶段的插穗基部皮层共鉴定到4 427种蛋白质,通过对同位素报告基团相对含量的定量检测,共检出2 875种蛋白质。其中,扦插初期和膨大期的差异蛋白质有595种,扦插初期和发根期的差异蛋白质有660种,膨大期和发根期的差异蛋白质有231种。从表达差异显著的蛋白质中筛选出GDP-D-甘露糖基-3’,5’-异构酶、凝集素KM+、生长素IAA水解酶、热激蛋白hsp70、丙二烯氧化物环氧酶、细胞色素b6、过氧化物酶等可能与生根过程密切相关的蛋白质,通过GO功能注释和KEGG数据库比对分析,这些差异蛋白质在扦插生根中主要执行的功能包括代谢过程、多细胞组织进程、定位系统建立、生物学调控、刺激应答、细胞要素、细胞器要素、催化活性和蛋白质结合等,并主要参与次生代谢产物的生物合成、淀粉和蔗糖代谢、内质网蛋白质加工、苯丙素生物合成、糖酵解和糖异生等代谢通路。
Differential proteome of base cortex at different rooting stages of mulberry hardwood cuttings were analyzed by applying isobaric tags for relative and absolute quantization（ iTRAQ） technology combined with liquid chromatographytandem mass spectrometry（ LC-MS/ MS） technology to provide basic information for studying regulatory mechanism of the rooting of mulberry cuttings at molecular level. Totally 4 427 proteins were identified in the base cortex of cuttings at the early stage after planting,expanding stage and rooting stage,wherein 2 875 proteins were identified by quantitatively analyzing the relative content of reporter groups marked with isotope. There were 595 differential proteins between the early stage after planting and expanding stage,660 differential proteins between the early stage after planting and rooting stage,and 231 differential proteins between the expanding stage and rooting stage,respectively. Proteins which may be closely related to rooting such as GDP-D-mannose-3 ＇,5 ＇-epimerase,lectin KM ＋,auxin IAA hydrolase,heat shock protein hsp70,allene oxide cyclase,cytochrome b6,and peroxidase were detected from the significant differential proteins. By contrasting and annotating with GO and KEGG databases,it was found that the above differential proteins mainly perform functions including metabolic process,multicellular organismal process,establishment of localization,biological regulation,response to stimulus,cell part,organelle part,catalytic activity and binding,etc. during rooting,and participate in various metabolicpathways such as biosynthesis of secondary metabolites,starch and sucrose metabolism,protein processing in endoplasmic reticulum,phenylpropanoid biosynthesis,glycolysis and gluconeogenesis,etc.
出处
《蚕业科学》
CAS
CSCD
北大核心
2015年第4期593-602,共10页
ACTA SERICOLOGICA SINICA
基金
现代农业产业技术体系建设专项(No.CARS-22)
关键词
桑树
硬枝扦插
生根
蛋白质组
同位素标记相对和绝对定量技术
液相色谱-串联质谱法
Morus L.
Hardwood cutting
Rooting
Proteome
Isobaric tags for relative and absolute quantitation technology
Liquid chromatography-tandem mass spectrometry technology
作者简介
杜伟（1985-），男，助理研究员，博士研究生。E—mail：yg123．456@126．com
通信作者信息：程嘉翎，研究员。E-mail：chengjialing@163．com
