苜蓿雄性不育植株与可育植株现蕾初期花蕾蛋白质组比较

采用双向凝胶电泳对现蕾初期苜蓿雄性不育植株(Ms-4)及其可育植株(MF)花蕾蛋白质进行了分离,获得了分辨率和重复性较好的双向电泳图谱。通过ImageMaster 2D软件对Ms-4和MF银染图谱分析发现,两者在等电点5~7、分子量20~60 kD范围内蛋白质斑点分布最多,可识别的总蛋白质点数均在6 000个左右,其中差异表达的蛋白质点数为98个;进一步通过质谱分析成功鉴定了22个差异蛋白点。利用Blast2GO程序对 22个蛋白点进行功能注释和代谢途径分析发现,核酮糖羧化酶小亚基、尿苷三磷酸-葡萄糖-1-磷酸尿苷酰基转移酶等蛋白在光合作用、碳水化合物代谢、多细胞生物有机体的发育等过程中起着重要的作用,同时参与了细胞质、细胞壁等组成,并具有绑定、催化、结合和水解等功能。研究结果初步推断,在苜蓿花药发育过程中,蛋白的缺失及表达量的变化可能会使与花粉发育有关的能量缺失,物质合成发生改变,导致雄性不育。

Comparative Proteomic Analysis of Bud Proteins of Alfalfa Male Sterile and Fertile Plants at Bud Emergence Stage

The dimensional electrophoresis profiles with good resolution and repeatability was got by separating the proteins of alfalfa on the bud of male sterile (Ms-4) and fertile plants(MF) of bud emergence stage.By using ImageMaster 2D software to analyze silver stained 2-DE gel,the protein spots of Ms-4 and MF mainly scattered in the isoelectric point of 5~7 with maximum distribution of molecular weight in 20~60 kD.There were about 6 000 expressed protein points in bud for Ms-4 and MF,and about 98 spots were differential expressed.The 22 protein spots were successfully identified by mass spectrometry analysis.The ribulose bisphosphate carboxylase small chain,UTP-glucose-1-phosphate uridylyltransferase and others played an important role in photosynthesis,carbohydrate metabolism and the development of multicellular organisms by using Blast2GO program to analyze function and metabolic pathway of the 22 proteins.Meanwhile,they were involved in the composition of cytoplasm,cell wall and other parts,which has some functions,including binding,catalytic,combining and hydrolysis and others.It concluded preliminary that the lack of protein and the changes of expression levels might lead to male sterility,which may attributed to the loss of energy and the change of material synthesis in the development process of alfalfa pollen.