摘要
春小麦是青海省的主要粮食作物,青海高原干旱频繁且严重,尤以春旱为首,对小麦生长发育造成严重影响.为了从蛋白质组学水平分析小麦对干旱胁迫的应答特征,探讨小麦可能的抗旱机制,本研究对青海主栽春小麦品种青春38幼苗进行聚乙二醇(polyethylene glycol,PEG)6000胁迫和复水处理,采用IEF/SDS-PAGE双向凝胶电泳技术,对PEG胁迫和复水处理的小麦叶片总蛋白质分别与正常浇水的对照进行差异蛋白质组学研究,经考马斯亮蓝G-250染色获得清晰度和重复性较好的双向电泳图谱.PD Quest软件处理分析各对照和处理图谱,在等电点4.0～7.0线性范围内,均可识别650个以上清晰蛋白质点,获得比较明显的差异表达蛋白点43个,其中8个蛋白点重复.从35个差异蛋白点中选取24个差异点进行MALDI-TOF-TOF-MS肽质量指纹图谱分析,应用Mascot软件在NCBInr数据库中搜索鉴定蛋白质,得到22个阳性结果.对鉴定得到的差异表达蛋白进行功能分析,它们分别参与了光合作用、蛋白质合成、能量代谢途径、细胞防御、氧化还原、运输、信号转导等过程,而且根据差异表达蛋白功能分类所占比例,发现干旱胁迫与光合作用关系最为紧密.
The severe drought frequently occurred in Qinghai China during severely interferes the wheat growth and development.To better understand drought resistance at the protein expression level,differential proteomic analysis was carried out in PEG-stressed and re-watered spring wheat（Qingchun38） young leaves.The seedlings with two leaves were treated with PEG6000 for 72 hours and re-watered for 24 hours,proteins were extracted,separated by IEF/SDS 2D gel electrophoresis and stained with Coomassie brilliant blue G-250.More than 650 reproducible abundant protein spots that were detected on each 2D-gel within pI 4~7 by PDQuest software.Of these proteins,43 of which varied quantitatively or qualitatively each 2D-gel in 8 sample repeats.A total of 24 differentially expressed proteins were subjected to mass spectrometry,22 of them were identified in the NCBInr database.The functional categories of the identified proteins were photosynthesis,protein biosynthesis,energy pathway,cell defense,oxidoreduction,transportation,signal transduction.The results also showed that PEG stress was strongly connected to photosynthesis.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2012年第11期1049-1056,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
青海省高原作物种植资源创新与利用重点实验室培育基地开放课题资助(No.2011-04)~~
关键词
春小麦
聚乙二醇6000胁迫
复水
差异蛋白质组学
基质辅助激光解吸电离飞行时间质谱
spring wheat
PEG6000 stress
re-watering
differential proteomics
Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry（MALDI-TOF-TOF-MS）
