Differential Proteomic Analysis Of The Arabidopsis Thanliana In Response To Insect Feeding And Mechanical Wounding

Insects feeding, which seriously affects the yield and quality of agricultural products, is one of the most important biological pressures to host-plants. In this research, the mechanism of herbivore-induced plant defense response was studied using proteomics technology, trying to figure out the function of related proteins.Based on the research system of specialist Plutella xylostella and the model plant Arabidopsis thaliana, the parameters of 2-DE for Arabidopsis thaliana was optimized and the proteomic research flat was succeeded constructed of interaction of Plutella xylostella and Arabidopsis thaliana. We used Tris saturated phenol to extract the total proteins from the whole plants. PVPP, which can absorb the plant phenols, was added in whetting by liquid N2, then we totally removed pigments, salts,and other impurities using different acetone concentrations and finally obtained high purified protein. Suitable loading sample is very important factor in affecting the resolution of 2-DE. Silver dyeing, 18cm IPG (pH4-7)and 300ug sample were used in our study, we obtained the better result, which has the clear background and protein spots and it is suitable for gel analysis.Changes of differential expressive proteins were demonstrated after insects feeding and mechanical injure. In the former, 52 protein spots were founded to be changed more than 2 folds in their expression abundance in average respectively, and of which 39 protein spots increased while 13 spots were down-regulated differently. In the latter, 44 protein spots were founded to be changed in their expression abundance in average respectively, and of which 21 protein spots increased and 23 spots were down-regulated differently. Only 24 protein spots changed the expression abundance in the Plutella xylostella feeding stress were identified by MALDI-TOF-TOF MS including Glycine-rich RNA-binding protein, ATP synthase subunit alpha, ATP synthase subunit beta, GTPase-activating protein, Glyceraldehyde-3-phosphate dehydrogenase, ADP-glucose pyrophoshorylase, Serine/threonine protein kinase, 2-Cys peroxiredoxin, Ribulose bisphosphate carboxylase, putative elicitor-responsive gene etc. These feeding responsive proteins are involved in signal transduction, proteins processing and energy metabolism etc, classified by the online databases and related references. Because of most of these proteins located on/in the cell membrane, we predicted that these proteins play important roles in driving signal pathway of herbivore-induced plant defense response.What we have done in this study makes the way for further research in insect-plants interaction. It will play important role in unfolding the heredity of herbivore-induced plant defense and in finding the specially expressed proteins in insects feeding stress.