甜菜丛根病抗性差异性状及蛋白胶点鉴定比较研究

Comparative Study on Differential Characters and Proteins Spot Identification of Resistance to Sugar Beet Rhizomania

以甜菜抗病和感病材料为试验材料,对田间产量进行了比较。通过比较蛋白质垂直电泳图谱,比较了TCA/丙酮沉淀法、可溶性蛋白提取法和酚提取法3种不同蛋白质提取方法,水培进行病毒胁迫处理后,通过Image Master 2D Platinum Trial 5. 0软件分析,扣除差异蛋白点,进行胶内蛋白质鉴定分析。结果表明:3种蛋白质提取方法中的TCA/丙酮沉淀法提取效果较好。田间产质量比较显示,抗病材料在苗期及生长后期长势要优于感病材料,收获后块根产量和含糖量也要高于感病材料,抗病材料表现良好。蛋白质双向电泳后通过软件分析,扣除具有明显差异的蛋白点78个,总共鉴定出了34个有效蛋白质ID,通过NCBI和The Beta vulgaris Resource数据库Blast比对发现,病1和病2主要表达的功能蛋白以光合作用、糖代谢、呼吸抗氧化作用、信号转导、脂代谢及一些未知蛋白为主,而病5和病6主要表达光合作用蛋白和呼吸抗氧化蛋白,即自身应激反应所表达的蛋白质。

Sugar beet resistant and susceptible materials were used to compare the yield and quality in the field.Three different methods of protein extraction were compared,which included TCA/acetone precipitation method,soluble protein extraction method and phenol extraction method.After the treatment of viral stress by water culture,the different protein spots were analyzed by Image Master 2D Platinum Trial 5.0 software.The results showed that TCA/acetone precipitation method had better extraction effect.The comparison of field yield and quality showed that the growth of resistant materials were better than that of susceptible materials at seedling stage and late growth stage,and the yield and sugar content after harvest were higher than those of susceptible materials,thus the resistant materials performed well.The protein was analyzed by Image Master 2D Platinum Trial 5.0 software after two-dimensional electrophoresis,and 78 protein spots with obvious difference were deducted.A total of 34 active protein IDs were identified.Compared by NCBI and The Beta vulgaris Resource database Blast,the main functions of proteins expressed by disease 1 and disease 2 were photosynthesis,glycometabolism,respiratory antioxidation,signal transduction,lipid metabolism and some others.While disease 5 and disease 6 mainly expressed photosynthesis protein and respiratory antioxidant protein which were the proteins expressed by self stress reaction.