摘要
为了阐明外源H2S对高盐胁迫下黄瓜蛋白质表达的影响,以盐敏感型黄瓜栽培种‘春夏秋王’为材料,通过双向电泳(2-DE)技术分离叶片总蛋白质,采用基质辅助激光解析飞行时间串联质谱(MALDI-TOF/TOF-MS)技术对差异表达蛋白质进行质谱鉴定,并利用NCBI及SwissProt数据库检索进行差异表达蛋白质功能注释和代谢通路分析。结果表明:(1)共检测到约2 490个蛋白质,其中蛋白质表达差异在1.5倍以上且差异显著(P<0.05)的有45个,其中有24个蛋白质表达上调,21个蛋白质表达下调。(2)成功鉴定蛋白质26个,这些蛋白质参与了光合作用(26.92%)、蛋白质代谢(23.08%)、能量与碳水化合物代谢(11.54%)、氨基酸生物合成(11.54%)、细胞结构相关蛋白(7.69%)、抗氧化作用(3.85%)、信号转导(3.85%)及未知功能蛋白(11.54%)。(3)GO注释表明差异表达的蛋白质分子功能主要以蛋白质结合与水解酶活性为主,生物学过程涉及应激反应、有机物代谢过程、胁迫响应和细胞分化等,细胞组成集中分布在细胞部分和一些胞内细胞器。KEGG代谢通路分析表明,差异表达蛋白质主要参与了肌动蛋白细胞骨架调控、细胞凋亡、碳代谢和光和调控等代谢途径。研究表明,外源H2S诱导了高盐胁迫下黄瓜叶片蛋白质的差异表达,为后续探索外源H2S对黄瓜的抗盐分子机制研究提供了理论依据。
The objective of this work was to elucidate the effect of exogenous H2 S on cucumber protein expression under high salt stress.The salt-sensitive cucumber cultivar’Chunxiaqiuwang’was used as experimental material.The leaf proteins were separated by two-dimensional electrophoresis(2-DE),and the differential proteins were identified by matrix-assisted laser desorption ionization tandem time-of-flight/time-of-flight mass spectrometry(MALDI-TOF/TOF-MS).Functional annotation and metabolic pathway analysis of differentially expressed proteins were performed using NCBI and SwissProt database searches.The results showed:(1)about 2 490 protein spots were detected,of which 45 were differentially expressed at 1.5-fold and P <0.05 levels.The 24 proteins were up-regulated,and 21 proteins were down-regulated.(2)26 proteins were successfully identified.These proteins are involved in photosynthesis(26.92%),protein metabolism(23.07%),energy and carbohydrate metabolism(11.54%),amino acid biosynthesis(11.54%),cell structure related protein(7.69%),antioxidant reaction(3.85%),signal transduction(3.85%)and unknown functional protein(11.54%).(3)The GO annotation showed that many differential expressed proteins is mainly composed of protein binding and hydrolase activity in molecular function,distributed in response to stimulus,organic substance metabolic process,response to stress and cell differentiation etc.in biological process,cell part and intracellular organelle in cellular component.KEGG metabolic pathway analysis showed that differential proteins are mainly involved in actin cytoskeleton regulation,apoptosis,carbon metabolism and light and regulation.The research indicated that exogenous H2 S induced differential expression of cucumber leaf protein under high salt stress,which provided a theoretical basis for the subsequent exploration of exogenous H2 S on the salt-resistant molecular mechanism of cucumber.
作者
任绪明
蒋景龙
孙旺
李丽
REN Xuming;JIANG Jinglong;SUN Wang;LI Li(School of Biological Sciences and Engineering, Shaanxi University of Technology, Hanzhong, Shaanxi 723001, China;College of Chemical and Environmental Sciences, Shaanxi University of Technology, Hanzhong, Shaanxi 723001, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第12期2236-2248,共13页
Acta Botanica Boreali-Occidentalia Sinica
基金
陕西省科技厅项目(2018NY-042)
陕西省教育厅项目(17JK0152)
陕西理工大学研究生创新基金(SLGYCX1830)
关键词
盐胁迫
双向凝胶电泳
H2S
GO注释
KEGG通路分析
聚类分析
salt stress
two-dimensional gel electrophoresis
H2S
GO annotation
KEGG pathway analysis
cluster analysis
