渗透胁迫诱导转Cu／ZnSOD和APX基因甘薯根系差异蛋白质组学研究  

Differential expression of root proteins in the Cu/Zn SOD and APX gene co-transgenic sweet potato to osmotic stress

作　　者：康乐[1,3] 李红兵[3] 陈显让[2] 郭尚洙 邓西平[3] 

机构地区：[1]西北农林科技大学林学院,陕西杨凌712100 [2]西北农林科技大学资源环境学院,陕西杨凌712100 [3]中国科学院水利部水土保持研究所/黄土高原土壤侵蚀与旱地农业国家重点实验室,陕西杨凌712100 [4]韩国生命工学院环境生物技术研究中心,韩国大田305-806

出　　处：《西北农林科技大学学报（自然科学版）》2013年第10期88-96,共9页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国际科技合作项目(2010DFA91930)

摘　　要：【目的】利用蛋白质组学手段,研究短期渗透胁迫条件下,转Cu/Zn SOD和APX基因甘薯(T)及非转基因甘薯(NT)根系蛋白质的表达差异。【方法】以水培转Cu/Zn SOD和APX基因甘薯(T)及非转基因甘薯(NT)根系为材料,用100g/L PEG6000渗透胁迫处理24h,以未胁迫的NT为对照,提取根总蛋白,进行双向电泳分离,考染法凝胶染色,扫描凝胶图像用于软件检测。【结果】在T、NT和对照中分别检测到836,812,881个蛋白点;t测验发现,与对照相比,T和NT有34个蛋白点丰度发生1.5倍以上显著变化(P<0.05,Fold>1.5,Quality>80)。经基质辅助激光解析飞行时间质谱(MALDI-TOF-MS)分析,34个差异表达蛋白点中23个得到了有效鉴定。在鉴定的23个蛋白中,有10个蛋白在T和NT中共同上调或下调表达,包括ATP合酶F1β亚基、烯醇酶、26S蛋白酶体调节亚基6、ACC合成酶2等;另外13个蛋白在T和NT中表达各异,包括核酮糖-1,5-二磷酸羧化酶/加氧酶结合蛋白β亚基和蔗糖合成酶等。【结论】10个在T和NT中共同上调或下调表达的蛋白,是甘薯应答渗透胁迫的普遍应答蛋白,反映了T和NT根系对短期渗透胁迫共同的响应机制;另外13个在T和NT中表达各异的蛋白,表明转基因使甘薯根系蛋白质在短期渗透胁迫下表现出特异的响应机制。【Objective】Proteomics methods were used to study the root protein expression differences of the Cu/Zn SOD and APX gene co-transgenic sweet potato （T） and non-transgenic sweet potato （NT） to short-term osmotic stress.【Method】The Cu/Zn SOD and APX gene co-transgenic sweet potato （T） and non-transgenic sweet potato （NT） were cultivated by hydroponics,and treated for 24 hours under 100 g/L PEG6000 penetration stress,followed by two-dimensional electrophoresis and CBB staining,and the gel images was scanned for software testing.No stress NT was the control （CK）.【Results】836,812 and 881 protein spots were respectively detected in T,NT and CK.Compared with CK,34 protein spots were significantly expressed in two treatments with 1.5 times through t-test （P〈0.05,Fold〉1.5,Quality〉80）.The 34 protein spots were analyzed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry,23 of them were effectively identified.In these 23 protein spots,10 were co-upregulated or co-downregulated in T and NT,including beta subunit of ATP synthase F1,enolase,26S proteasome regulatory subunit 6 and ACC synthase,whereas 13 were differentially expressed in T and NT,including ribulose-1,5-bisphosphate carboxylase/oxygenase binding protein beta subunit and sucrose synthase.【Conclusion】10 co-upward or co-downward expression proteins were generally responsive proteins to short-term osmotic stress in sweet potato,reflecting a common response mechanism of T and NT roots in shortterm osmotic stress.Additionally,other 13 different expression proteins in T and NT indicated that the roots proteins of transgenic sweet potato presented specific performance in the short-term osmotic stress response mechanism.

关 键 词：转基因甘薯 蛋白质组学 双向电泳 基质辅助激光解吸飞行时间质谱 

分 类 号：S531.01[农业科学—作物学] Q945.78[生物学—植物学]