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Characterization of Proteins Responsive to Gibberellin in the Leaf-Sheath of Rice (Oryza sativa L.) Seedling Using Proteome Analysis

机译:蛋白质组分析法鉴定水稻幼苗叶鞘中对赤霉素的响应蛋白

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In plants gibberellins (GAs) are responsible for triggering stem or internodal elongation. To comprehend the molecular basis of internodal elongation in rice, a proteomics approach using differentially displayed proteins on two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was carried out to identify the proteins expressed during the GA controlled leaf-sheath elongation response. Out of 352 protein spots detected on 2-D PAGE, 32 proteins showed modulation in the expression levels in GA_3-treated leaf-sheath for 48 h as compared to control. These proteins were analyzed using protein sequencer and/or mass spectrometry in conjunction with the protein database to assign putative identities. The twin protein spots (LS079 and LS083), identified as calreticulin, showed different isoelectric points and expression level in GA_3-treated leaf-sheath. The expression level of LS083 (pI 4.0) was down-regulated as compared to the up-regulation of LS079 (pI 4.3). In the presence of GA_3 and growth inhibitor, uniconazole and abscisic acid, respectively, no elongation in leaf-sheath occurred and calreticulin did not shift from LS083 to LS079. Over-expression of calreticulin in rice inhibited the callus regeneration and seedling growth. These results suggest that calreticulin is an important component in the GA signaling pathway that regulates rice seedling leaf-sheath elongation.
机译:在植物中,赤霉素(GA)负责触发茎或节间伸长。为了理解水稻节间伸长的分子基础,采用蛋白质组学方法,在二维聚丙烯酰胺凝胶电泳(2-D PAGE)上使用差异显示的蛋白质进行了鉴定,以鉴定在GA控制的叶鞘伸长反应中表达的蛋白质。在2-D PAGE上检测到的352个蛋白质斑点中,与对照组相比,GA_3处理的叶鞘中的32个蛋白质表达水平有所调节。使用蛋白质测序仪和/或质谱分析法结合蛋白质数据库对这些蛋白质进行分析,以分配假定的身份。鉴定为钙网蛋白的孪生蛋白质斑点(LS079和LS083)在GA_3处理的叶鞘中显示出不同的等电点和表达水平。与LS079(pI 4.3)的上调相比,LS083(pI 4.0)的表达水平下调。在存在GA_3和生长抑制剂烯效唑和脱落酸的情况下,叶鞘均未发生伸长,钙网蛋白也未从LS083转变为LS079。钙网蛋白在水稻中的过量表达抑制了愈伤组织的再生和幼苗的生长。这些结果表明钙网蛋白是调节水稻幼苗叶鞘伸长的GA信号通路中的重要组成部分。

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