低磷胁迫下油菜幼苗叶片蛋白酶二维酶谱分析

Differential Zymogram Pattern of Protease in Leaves of Brassica napus Seedlings under Low Phosphorus Stress

为探讨低磷胁迫对油菜(Brassica napus)幼苗叶片蛋白酶二维酶谱的影响,以低磷胁迫下的甘蓝型油菜鼎油杂4号为试材,采用优化的非变性双向电泳技术结合蛋白酶谱检测技术获得清晰的蛋白酶分离图谱,通过图像分析获得磷胁迫下油菜叶片蛋白酶差异表达图谱。结果显示,低磷胁迫下共有6个油菜叶片蛋白酶点表达水平具有显著差异(表达量差异在2倍以上),其中3个表达水平显著上调,3个表达水平显著下调。结果提示有不同的油菜叶片蛋白酶类参与到油菜抗(耐)低磷的应答途径中,为进一步解析不同蛋白酶(同工酶)响应低磷胁迫研究打下基础。文中基于非变性等电聚焦的二维酶谱方法亦为其他逆境胁迫下不同蛋白酶及其同工酶二维酶谱响应研究提供技术参考。

To investigate the effect of low phosphorus stress on the expression of protease zymogram content in seedling leaves of Brassica napus,Dingyouza No.4 was used as experimental material.The seedlings were subjected to phosphorus deficiency nutrient solution for 10 days under low phosphorus stress,and leaf proteins were extracted by direct grinding method.Optimized non-denaturation two-dimensional electrophoresis combined with protease detection technology was used to obtain a good protease separation map.Protein difference analysis was performed using ImageMaster 2D Platinum 7.0 software.The results showed that the expression levels of 6 protease points in rape leaves were significantly different under low P stress(the difference was more than 2 times),among which 3 were significantly up-regulated and 3 were significantly down-regulated.The results suggested that different leaf proteases are involved in the response pathway of rape to low phosphorus.The two-dimensional zymogram technology based on non-denaturing two-dimensional electrophoresis mentioned in this paper also laid a foundation for the study of protease zymogram.