‘培矮64S’与其eui突变体‘长选3S’穗颈节间蛋白质组差异分析  

Proteomics Difference Analysis of Elongated Uppermost Internode between TGMS Rice Line‘Peiai 64S’and Its eui Mutant‘Changxuan 3S’

作　　者：肖辉海 郝小花 XIAO Huihai;HAO Xiaohua(Department of Life Science, Hunan University of Arts and Science, Changde 415000, Hunan, China)

机构地区：[1]湖南文理学院生命与环境科学学院,湖南常德415000

出　　处：《西北植物学报》2020年第6期957-968,共12页Acta Botanica Boreali-Occidentalia Sinica

基　　金：湖南省自然科学基金(12JJ2018);湖南省重点学科建设项目(湘教发[2011]42号)。

摘　　要：为从蛋白质表达水平了解长穗颈温敏核不育水稻穗颈节间伸长机理,该研究以长穗颈(EUI)温敏核不育水稻‘长选3S’为材料,温敏核不育水稻‘培矮64S’为对照,采用固相pH梯度双向凝胶电泳和质谱分析方法,对2个水稻材料抽穗前2 d的穗颈节间蛋白质进行分离,并进行差异蛋白质组学的比较研究。结果表明:(1)获得了分辨率和重复性较好的双向凝胶电泳图谱。(2)对40个差异蛋白质点进行MALDI-TOF-TOF-MS肽质谱指纹图谱分析,成功鉴定其中27个差异蛋白质点;与‘培矮64S’相比,‘长选3S’中有17个上调表达和10个下调表达的蛋白质。(3)差异蛋白质按照其功能可分为6类,其中主要是与细胞代谢相关蛋白,其次是与细胞壁重建相关蛋白;并且这些差异蛋白质可能与‘长选3S’抽穗期穗颈节间剧烈伸长生长有关,尤其是细胞壁重建相关蛋白与细胞的伸长密切相关。(4)实时荧光定量PCR对随机挑选的蛋白点2、7、8、24、35和36所对应的基因在两个材料最上节间的表达结果显示,‘长选3S’的2(Os10g08550)、7(Os12g42876)、8(Os01g55830)基因的表达量较‘培矮64S’明显下调,而24(Os06g48760)、35(Os05g25850)、36(Os07g42300)基因的表达量较‘培矮64S’显著上调,表明q-PCR的结果与蛋白凝胶图分析结果一致。研究认为,水稻eui基因可能是通过调节抽穗期穗颈节间这些蛋白质的表达,从而促进穗颈节间细胞分裂,尤其是细胞的伸长生长。In order to understand the mechanism of neck-panicle elongation in protein level,we used‘Changxuan 3S’and‘Peiai 64S’as experimental materials.The proteins in the uppermost internodes were separated on the 2 day before heading by using solid-phase pH gradient two-dimensional gel electrophoresis and mass spectrometry.A two-dimensional gel electrophoresis map(2-DE)with good resolution and reproducibility was obtained.And the different proteins were studied.The results showed that:(1)two dimensional gel electrophoresis maps with good resolution and reproducibility were obtained.(2)40 differential protein spots were selected from the 2-DE map and analyzed by MALDI-TOF/TOF-MS peptide mass fingerprinting,and 27 of them were identified successfully.Compared with‘Peiai 64S’,there are 17 up-regulated and 10 down-regulated proteins in‘Changxuan 3S’.(3)According to their functions,these differential proteins can be divided into six categories,mainly including cell metabolism relative proteins and cell wall reconstruction proteins.In addition,these discrepant proteins may be interrelated to the vigorous elongation of the internodes at heading stage of‘Changxuan 3S’,especially the proteins related to cell wall reconstruction and cell elongation.(4)The protein spot 2,7,8,24,35 and 36 were tested by q-PCR.The expression levels of 2(Os10g08550),7(Os12g42876)and 8(Os01g55830)in‘Changxuan 3S’were significantly lower than that in‘Peiai 64S’,while the expression levels of 24(Os06g48760),35(Os05g25850)and 36(Os07g42300)were significantly higher than those of‘Peiai 64S’.The q-PCR results were consistent with the results of protein gel analysis.According to the study,the eui may promote cell division and elongation by regulating the expression of these proteins at heading stage.

关 键 词：温敏核不育水稻 长穗颈突变体 穗颈节间 蛋白质组学 

分 类 号：Q786[生物学—分子生物学] Q789