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采用固相pH梯度SDS聚丙烯酰胺双向凝胶电泳技术对杀雄剂SQ-1处理和未处理的小麦(Triticum aestivum)单核期、二核期花药总蛋白进行了分离,通过考马斯亮蓝G-250染色,获得了分辨率和重复性较好的双向电泳图谱。PDQuest2DE图像分析软件分析结果表明:(1)在等电点(pI)4～7之间可识别约500个以上较为清晰的蛋白质点,处理和对照各时期蛋白质在pI5～6,分子量20～40kD之间相对较集中。(2)检测到单核期差异点43个,二核期差异点45个,计算各差异点的分子量和pI。单核期43个差异点中,11个点在处理材料的单核期缺失而在对照中表达,7个在处理中表达而在对照中缺失;14个点表达量在处理中明显减弱,而另外11个明显增强。(3)二核期45个差异点中,有13个在处理中缺失而在对照中表达,7个在处理中表达而在对照中不表达;12个表达量在处理中明显减弱而另外13个明显增强。(4)经SQ-1处理后在单核期和二核期A-Z3(26.8/5.7)和C-Z5(26.8/5.7)点均缺失;点A-L10(26.8/5.9)在单核期处理的表达量下调,而到了二核期完全缺失C-Z6(26.9/5.9)点。通过双向电泳技术所获得的这些差异蛋白很可能直接或间接地参与了供试材料花药正常发育与败育发育的某些关键途径,而处理和对照互为等生理差异系,两者图谱中表现出的差异蛋白又很可能与SQ-1诱导小麦雄性不育有关。
Solid phase pH gradient SDS polyacrylamide gel electrophoresis was used to separate the mononuclear and dinuclear anther total proteins of the SQ-1-treated and untreated wheat (Triticum aestivum). Coomassie brilliant blue G -250 staining, obtained a better resolution and repeatability of two-dimensional electrophoresis. The analysis results of PDQuest2DE image analysis software showed that: (1) About 500 clear protein spots were identified between the isoelectric points (pI) of 4 ~ 7, and the proteins at pI5 ~ 6 and the molecular weight of 20 ~ 40kD Relatively concentrated between. (2) There were 43 differential points in mononuclear stage and 45 different points in dinucleate stage, and the molecular weight and pI of each difference point were calculated. Eleven of the 43 single-nucleus differences were expressed in the control at the mononuclear phase of the treated material, 7 were expressed in the treatment but were absent in the control, and the 14-point expression was significantly weakened in the treatment The other 11 were significantly enhanced. (3) Of the 45 differences in the binucleate stage, 13 were deleted in the treatment but were expressed in the control, 7 in the treatment but not in the control; 12 were significantly weakened in the treatment and 13 A significant increase. (4) Deleted at SQ-1 at A-Z3 (26.8 / 5.7) and C-Z5 (26.8 / 5.7) Decreased expression of the stage of treatment, and the complete deletion of C-Z6 (26.9 / 5.9) point at the binuclear phase. These differential proteins obtained by two-dimensional electrophoresis are likely to be directly or indirectly involved in some of the key pathways for the normal development and abortion of the anther of the tested materials, whereas the physiological differences between the treatments and the controls are of the same type. The difference protein is likely to be related to SQ-1-induced wheat male sterility.