Domestication-related Proteins In Soybean: A Proteomic Approach

1.Optimizations were taken on soybean leaves proteomics two-dimensional gel electrophoresis in aspects of protein sample preparation,sample loading volume and IEF procedure:Optimized TCA/acetone method can promote protein resolution; sample loading volume was increased to 450μg which makes better display of low-peak protein spots;prolonged time of low voltage desalting step was modulated in IEF procedures to get better focusing results.With optimized method,using Glycine max and Glycine soja leaves as materials,high repeatable results of two-dimensional gel electrophoresis were obtained,gel matching results by using ImageMaster software show,both the match rate of intra-group and inter-group are more than 80%.2.Soybean leaves two-dimensional gel electrophoresis experiments was taken using materials of Glycine max and Glycine soja population mixed sample.Gels were analysed by ImageMaster software,668 reliable protein spots in average were found. All matched spots were tilted by Ratio＞1.5 and T test analysis(P＜0.05) to find significant proteins:96.55%of total proteins have no significant difference;23 significant protein were found,accounting for 3.45%of total proteins.12 proteins have higher expression volumes in Glycine max,and 11 proteins lower,accounting for 1.80%and 1.65%,respectively.In addition,after matching by ImageMaster software and naked eye,no specific protein was found neither in Glycine max nor in Glycine soja group.Assume that differences brought by the domestication reflects on protein expression volume.3.10 main significant proteins were identified by MALDI-TOF-MS and matched in NCBInr protein database,all got positive searching results(p＜0.05, Score≥70),are:Serine hydroxyl- methyltransferase[Glycine max],thioredoxin H [Populus tremula×Populus tremuloides],acyl desaturase[Brassica rapa subsp. pekinensis],Gamma-glutamyl hydrolase precursor[Glycine max],Stem 31 kDa glycoprotein precursor[Glycine max],Stem 28 kDa glycoprotein precursor[Glycine max].granule-bound starch synthaseⅠ[Viburnum sargentii],zeta-carotene desaturase [Narcissus tazetta var.chinensis],hypothetical protein[Oryza sativa Japonica Group].Most of them have close relation with soybean domestication.