Research On The Changes Of Proteome In The Seed Embryo Of Zea Mays During The Artificial Aging

Two-dimensional electrophoresis with high resolution and sensitivity can be used to effectively isolate proteins in tissue or cells. It has a huge advantage in understanding the whole picture of cell activity. Consequently, it has become the core technology in complex protein mixture separation. In this paper, by using the maize variety"Dabaitou"as material, the changes of maize embryos proteome under artificial aging conditions were studied by means of two-dimensional electrophoresis and peptide mass fingerprinting. Three different energy levels of the"Dabaitou"embryo proteins had been successfully isolated by two-dimensional electrophoresis. The main results are as follows:1. Good protein samples were obtained by TCA-acetone precipitation method, which could meet the requirements of two-dimensional electrophoresis. Comparion with sample volume of 0.8mg/L, 500μL system, a sample volume of 0.8mg/L, 350μL system is better, that is less horizontal stripes, clearer and more protein spots can be obtained in two-dimensional electrophoresis, under the conditions of with 17cm, pH3-10 linear IPG strip, stained by Coomassie brilliant blue, and the recommended instrument parameters.2. Under the sample volume of 0.8mg/L, 350μL system, we separated the zea embryo protein of 3 defferent viability levels with the control under the above two-dimensional electrophoresis system. Using PDQuest 8.0.1 software for image analysis, more than 400 valid protein points were found in the gel map, and the proteins which had 2-fold different expression were identified by mass spectrometry, 22 proteins were identified successfully.3. The identified 22 proteins could be divided into 7 categories according to their function and metabolic pathwaysⅠ: disease / defence (5);Ⅱ: protein destination and storage (4);Ⅲ: transcription (3);Ⅳ: anabolic (1);Ⅴ: energy (3);Ⅵ: cell growth / division (2);Ⅶ: unknown protein (4).Therefore, a great change of proteome in the seed embryo have taken place, it affects many facets of seed vigor, such as energy metabolism system, cell growth / division, the synthesis of biological macromolecules in cells, defense system and so on.