The change of glutamine synthetase (GS) isoform was determined by Native PAGE and the activity staining as well as Western blotting. The activity changes of GS, NADH dependent glutamate synthase (NADH GOGAT), and NADH...The change of glutamine synthetase (GS) isoform was determined by Native PAGE and the activity staining as well as Western blotting. The activity changes of GS, NADH dependent glutamate synthase (NADH GOGAT), and NADH dependent glutamate dehydrogenase (NADH GDH) were also analyzed by enzyme assay. The results indicateded that only one form of GS isozyme was detected during the germination of cushaw seed and the different development stages of the cotyledon grown in light or in darkness. GS activity was increaseed with the germination of the seed untill the cotyledon to begin green and then was reduced gradually. The tendency of the activity change of NADH GOGAT was similar to GS. The activity change of NADH GDH was contrary to that of GS and NADH GOGAT. The results suggested that the expression of GS isoform was not effected by the treatments of light and darkness in the experiment conditions. GS/GOGAT cycle may play a main role for assimilating ammonium during the germination of the seeds and the initial stage of the cotyledon development, and the cycle was weaken and GDH pathway was strengthened after the green of the cotyledon by comparing these enzyme activities.展开更多
采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中...采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中差异表达的蛋白质点数为85。将其中16个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定。YTA相对于YTB有部分参与碳代谢和淀粉合成的酶缺失或表达量降低,这些蛋白质分别是ADP-葡萄糖磷酸转移酶(AGPase),UDP-葡萄糖醛酸脱羧酶,乙酰辅酶A合成酶和二氢硫辛酸脱氢酶等。其中AGPase是参与淀粉合成的蛋白,与花粉发育密切相关。乙酰辅酶A合成酶和二氢硫辛酸脱氢酶是细胞内合成乙酰辅酶A的重要酶,而乙酰辅酶A是进入TCA循环的重要底物,乙酰辅酶A的缺乏可以导致TCA循环不能顺利进行,从而不能提供小孢子发育所需要的大量能量。YTA相对于YTB部分参与碳水化合物代谢的重要酶缺失或表达量降低,有可能导致因线粒体提供的能量不足,淀粉合成受阻,因而花粉不能正常发育。展开更多
文摘The change of glutamine synthetase (GS) isoform was determined by Native PAGE and the activity staining as well as Western blotting. The activity changes of GS, NADH dependent glutamate synthase (NADH GOGAT), and NADH dependent glutamate dehydrogenase (NADH GDH) were also analyzed by enzyme assay. The results indicateded that only one form of GS isozyme was detected during the germination of cushaw seed and the different development stages of the cotyledon grown in light or in darkness. GS activity was increaseed with the germination of the seed untill the cotyledon to begin green and then was reduced gradually. The tendency of the activity change of NADH GOGAT was similar to GS. The activity change of NADH GDH was contrary to that of GS and NADH GOGAT. The results suggested that the expression of GS isoform was not effected by the treatments of light and darkness in the experiment conditions. GS/GOGAT cycle may play a main role for assimilating ammonium during the germination of the seeds and the initial stage of the cotyledon development, and the cycle was weaken and GDH pathway was strengthened after the green of the cotyledon by comparing these enzyme activities.
文摘采用固相pH梯度-SDS PAGE双向电泳对红莲型细胞质雄性不育水稻的不育系(YTA)和保持系(YTB)单核期花粉总蛋白质进行了分离,通过银染显色,获得了分辨率和重复性较好的双向电泳图谱。Image Master 2DV5.0软件可识别约1800个蛋白质点,其中差异表达的蛋白质点数为85。将其中16个差异点采用基质辅助激光解析电离飞行时间质谱(matrix assisted laser desorption/ionizaton time of flight mass spectrometry,MALDI-TOF-MS)进行了肽质指纹图分析,通过采用Mascot软件对MSDB数据库查询,其中9个蛋白质点得到了鉴定。YTA相对于YTB有部分参与碳代谢和淀粉合成的酶缺失或表达量降低,这些蛋白质分别是ADP-葡萄糖磷酸转移酶(AGPase),UDP-葡萄糖醛酸脱羧酶,乙酰辅酶A合成酶和二氢硫辛酸脱氢酶等。其中AGPase是参与淀粉合成的蛋白,与花粉发育密切相关。乙酰辅酶A合成酶和二氢硫辛酸脱氢酶是细胞内合成乙酰辅酶A的重要酶,而乙酰辅酶A是进入TCA循环的重要底物,乙酰辅酶A的缺乏可以导致TCA循环不能顺利进行,从而不能提供小孢子发育所需要的大量能量。YTA相对于YTB部分参与碳水化合物代谢的重要酶缺失或表达量降低,有可能导致因线粒体提供的能量不足,淀粉合成受阻,因而花粉不能正常发育。
