Proteome Analysis Of Arabidopsis Thaliana Induced By Plutella Xylostella Feeding

Nowadays, the research about herbivore-induced plant defense response using proteomics technology is becoming an important and hot topic in the the interaction study area between the insects and their host plants. In this study, Plutella xylostella and the model plant Arabidopsis thaliana were chosen as the research system and the comparative proteomic technology 2 DE was employed to explore the interactions of the insects feeding and plants defense response.Protein preparation and suitable protein buffer are very important factors in the plant proteomics research. At the very beginning, a suitable protein extraction method of Arabidopsis proteins for the two-dimensional electrophoresis was essential needed conditions for the next step work. In the protein extraction step, we used Tris-phenol to extracted the total proteins from the whole plants and then dissolved the mixture in the lysis buffer which contain TECP, SB3-10 to obtain high purified protein. More than 1000 protein spots were detected on each gel reproducibly by the ImageMaster? 2D Platinum 6.0 analysis software and the spots matching rate of each gel are more than 87%.Proteomic analysis was carried out to explore the the interaction of Plutella xylostella and Arabidopsis (Arabidopsis thanliana, ecotype Col-0). Changes of differential expressive proteins were demonstrated by the high-throughput MALDI-TOF-TOF MS aiming to examine induced abundance changes of proteins after 8h feeding after that about 30%-40% Arabidopsis leaves were removed by the third instar larvae. 38 protein spots were found to exhibit different dynamic patterns on 2-D gels in responding to the feeding stress of which 28 protein spots were detected to be up-regulated expressed while 10 of them were decreased in expression abundance respectively. 35 proteins were identified by MALDI-TOF-TOF MS successfully and it showed that some Metabolism-related enzymes were indentified, such as heat shock protein 70, Transketolase, malate dehydrogenase, adenylosuccinate synthase, beta-glucosidase, coproporphyrinogen III oxidase, glutathione S-transferase, acyl-CoA oxidase, etc., there were some unknown protein either.Experiments were also be carried out to explore the relationship between induced defense metabolism of plants and the jasmonic acid signaling pathway. 28 protein spots were founded to be changed more than 1.5 fold in their expression abundance in average repectively, and of which 19 protein spots increased in the relative vol% whist 9 spots were down-regulated differently. It is very interesting that there were 11 protein spots changed the expression abundance both in the feeding stress and the MeJA induced trait. 27 proteins were identified by MALDI-TOF-TOF MS including heat shock protein 70, profilin, glutamate-1-semialdehyde 2,1-aminomutase, adenylosuccinate synthase, methionine synthase, plastocyanin, pyrophosphorylase, etc., were identified in this trait.These feeding responsive proteins are involved in regulation of metabolism, energy, cell defense, biogenesis of cellular component, etc. classified by the NCBI, MapMan, TAIR databases and related references.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.