盐胁迫下棉花叶片差异蛋白表达的分析

以耐盐的陆地棉品种中07为研究材料,在其生长的三叶期,用未处理和0.4%NaCl胁迫处理24h的幼苗,分别提取其叶片全蛋白。通过等电聚焦和第二向SDS-PAGE技术获得完整的棉花叶片总蛋白图,利用ImageMaste-2DElite5.0软件分析各个差异蛋白在对照和0.4%NaCl胁迫24h后棉花叶片中的相对表达量,并选取质量好、实验重复性高的10个特异的差异表达蛋白质点进行基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF-TOF/MS)鉴定。结果表明,从对照和盐胁迫处理的双向电泳图谱中共检测到24个差异蛋白质点,这些差异蛋白质点的等电点分布集中在5.0～6.5之间,分子量分布集中在40.0～110.0kD之间;进一步质谱(MALDI-TOF-TOF/MS)分析鉴定的10个差异蛋白质点,其中4个蛋白质点获得了鉴定结果,这4个差异表达蛋白质点分别为1,5-二磷酸核酮糖羧化/氧化激酶α2(编号4)、1,5-二磷酸核酮糖羧化酶/氧化酶大亚基(编号5)、未知蛋白产物(编号19)和OEE2(编号32)。我们初步推测OEE2和1,5-二磷酸核酮糖羧化酶/氧化酶可能与棉花耐盐胁迫特性有关。

Analysis of Differential Proteins of Cotton Leaves Under Salt Stress

The salt tolerant upland cotton variety Zhong07 was employed as experimental materials,Leaves of trefoil seedlings treated with 0.4% NaCl.24 hours were sampled for whole protein extraction and the leaves of untreated cotton seedlings used as the control.Whole protein profiling of cotton leaf was obtained through isoelectric focusing technique and following SDS-PAGE.In this research we analyzed the relative abundance of differential proteins between control and stress treated cotton leaves by using software of ImageMaste-2D Elite(ver.5.0),and then we chose 10 differentially expressed protein spots that were high quality and reproducible to conduct analysis of MALDI-TOF-TOF/MS.The results showed that 24 differential proteins were detected for both of the control and salt-stress treatment,whose PI values were mainly concentrated in the range of 5.0~6.5 and molecular weight in the range of 30.0~110.0 KD.Ten differential proteins were chosen for further MALDI-TOF-TOF/MS analysis,of which four protein spots were successfully identified and annotated,namely,Ribulose-1,5-bisphosphate carboxylase/oxygenase activase alpha2(number 4),ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit(number 5),unknown protein product(number 19) and OEE2(number 32).We predicted that OEE2 and ribulose-1,5-bisphosphate carboxylase/oxygenase might involve in tolerance of salt stress in cotton.