A Two-Dimensional Electrophoresis Protocol to Interrogate Protein Discrepancies in Mesocarp and Putamen During Prunus persica Fruit Development

In order to realize the differentiated mechanism of mesocarp and endocarp in stone fruits, the profiling of protein expression in mesocarp and putamen of peach were analyzed by two-dimensional electrophoresis (2-DE) to explore the whole cellular milieu of proteomic variety. Peach fruits around 60 days after anthesis were adopted in this observation. The total protein was extracted by TCA/ cold acetone precipitation, and isolated by immobilized pH gradient (IPG) based on 2-DE routine. After silver staining, the gels evaluated operating image analysis software. Considerable high resolution and reproducible 2-DE patterns were obtained successfully. 1273plusmn101 and 1292plusmn115 protein spots were detected respectively from 2-DE patterns of mesocarp and putamen that pH 5~8 IPG strips were practiced in the process of isoelectric focusing. 65 spots of aggregate matched protein spots were present in both patterns of mesocarp and endocarp, and 30 spots showed 2-fold(t-test, p<0.05) up-regulated on quantitative appearing an increase in spot intensity and maximum abundance in putamen than mesocarp, in addition 35 protein spots expressed 2-fold(t-test, p<0.05) down-regulated. 10 spots were just observed in mesocarp´s pattern, whereas 18 spots only found in the pattern of putamen. The examination results suggest that the proteomic component in mesocarp and endocarp of peach during fruit development took on some discrepancies, and these diversities may be provide an explanation and foundation data ulterior about different direction of metabolic models in mesocarp and endocarp.