抗、感绿豆象绿豆种子差异蛋白的分离与鉴定

为寻找抗绿豆象绿豆种子中的重要抗虫成分,以抗虫绿豆晋绿7号、B20及感虫绿豆潍绿2117为试验材料,采用蛋白质双向电泳(2-DE)和质谱技术,对抗、感虫绿豆中差异蛋白及功能进行了鉴定与分析。结果表明,抗、感虫绿豆中差异蛋白表达量超过2.5倍的点共有15个。其中6个蛋白点通过数据库得到了成功鉴定,涉及3种蛋白质,分别为8S球蛋白(α亚型和β亚型)、核酮糖1,5-二磷酸羧化酶/加氧酶(RuBis CO)亚基结合蛋白和合成淀粉酶与胰蛋白酶两种抑制剂的前体多肽链。蛋白点B49(即8S球蛋白α亚型)和B31(RuBis CO亚基结合蛋白)在抗、感虫绿豆中的差异表达量分别达到10 000倍和23倍。抗虫绿豆中8S球蛋白α亚型及β亚型、RuBis CO伴侣蛋白及胰蛋白酶抑制剂的前体作为抗虫物质影响了绿豆象的生长发育甚至导致其死亡,与抗虫性的量效关系及联合效应还需进一步验证。

Separation and identification of differential protein in the insect-resistant and insect-susceptible mungbean

In order to find the insect-resistant composition and differentially expressed proteins of mungbean, Jinlv No.7, B20 and Weilv 2117 were used as experimental materials, and the differential proteins and functions of mungbean varieties that are resistant and susceptible to bruchids were compared and analyzed by two dimensional gel electrophoresis (2-DE) and identified by mass spectrometry. Among the samples, 15 protein spots showed a more than 2.5 times reproducible up-regulated, significance 6 of them were successfully identified by the database, and involved three kinds of proteins. They are the alpha and beta subtype 8S globulin, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBis CO) subunits binding protein and the precursor peptide chains for amylase inhibitor and trypsin inhibitor. The protein expressions of B49 (alpha subtype 8S globulin) and B31 (RuBis CO subunits binding protein) of insect-resistant mungbean were 10 000 and 23 times higher than that of insect-susceptible mungbeans. It stunted the growth and even death of the Callosobruchus chinensis L. that alpha and beta subtype 8S globulin and RuBis CO subunits binding protein and precursor peptide chains for amylase inhibitor and trypsin inhibitor of insect-resistant mungbean, the bruchid resistance effect of these three proteins need to further verified in terms of the quantity and the combined effect.