Proteome Analysis Of Arabidopsis Thaliana In Response To Spodoptera Exigua Feeding

Proteomics research in plant, the affecting factors of isolate production and a suitable protein buffer are important. To improve the downstream level of two-dimensional gel electrophoresis analysis has been urgently needed breakthrough technology issues, because of the high dynamic range and diversity, protein extraction method of analysis and comparison, the optimization of two-dimensional electrophoresis of Arabidopsis proteins is essential conditions. In the majority of plant material, rich pigment, phenol, lipids, polysaccharides such as interfering substances, Thus more difficult to obtain purified protein, Their existence weakened the IPG strip isoelectric focusing on protein absorption, This to a certain extent, affected the protein detection, In this study, the 28-day training Arabidopsis (ecotype Col-0) as experimental material, TCA-acetone, Tris-phenol, PEG precipitation three kinds of on the extraction method leaves of Arabidopsis thaliana isolate production and in the SDS polyacrylamide gel electrophoresis resolution in areas such as a comparative analysis, With a view to exploring an Arabidopsis proteins suitable for the analysis of the extraction methods.Study found that, Whether or protein isolate productionand SDS polyacrylamide gel electrophoresis resolution of Tris- phenol extraction are relatively better extraction methods. Tris- phenol protein extraction rate is twice the other two methods. Using Tris- phenol extraction was precipitated protein, Then dissolved reductant and gender containing liquid electrolyte solution dissolved, Get high isolate productionand and high protein electrophoresis resolution of the total sample, Suitable for two-dimensional gel electrophoresis analysis of the downstream operation. This protocol was developed using Arabidopsis as a source of protein and thus may also be applicable to protein preparations of other plants.A proteomic analysis was carried out aiming at understanding of the molecular adaptation mechanisms of Spodoptera exigua feeding stress in Arabidiopsis (Arabidopsis thanliana, ecotype Col-0). The seedlings were exposed to Spodoptera exigua feeding for 3 h respectively, together with the controls which were grown at normal. In PEG fractionation, the whole-cell proteins were extracted from both control and stressed seedlings. 28 protein spots were found to exhibit different dynamic patterns on 2-D gels in responding to Spodoptera exigua feeding stress and 21 of them were identified using MALDI-TOF MS. These feeding responsive proteins, besides three proteins of unknown function, are involved in regulation of metabolism, energy, cell defense, biogenesis of cellular component, etc. by MIPS function category, TAIR database and related references. Proteins (47%) were predicted to be located in the chloroplasts, Proteins (29%) in the cytoplasm and Proteins (14%) in the nucleus implying that chloroplast proteins are likely involved in the cold stress adaptation in Arabidopsis. The physiological implications, which were derived from the experimental data, are discussed, suggesting that there is a complex metabolic network in plants responsive to feeding stress.