The Research Of Differential Protein From Fokienia Hodginsii Under Salt Stress

Fokieniahodginsii is tree can grow in the poor soil with good wood and ornamental value as a garden tree under state-protected(category II)and native in FUJIAN province.Salt stress is one of the major nonbiological factors affecting plant growth and the geographical distribution.Proteomic techniques are effective approaches that have in recent years been adopted to study the mechanism of plants response to salt stress and to discover salt responsive proteins.it is meaningful for Fokieniahodginsii salt tolerant mechanism.this research use two-dimensional electrophoresis(2-DE),matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS)to study the salt stress for Fokieniahodginsii.first is built a system for(2-DE).three different methods including TCA/acetone,phenol extraction method and TCA/phenol extraction method were tested in the experiment.Besides,three different IPG strips of pH3-10,pH5-8,pH4-7 combined with three different focusing parameter which showed 20 000VH,25000VH,30 000VH respectively were performed in IEF.Finally,different concentration separation gel(10%,12.5%)were performed in all SDS-PAGE trial.The results indicated that by more protein spots were detected on 2-DE gels by phenol extraction method,IPG strips of pH 4-7,25000VH focusing parameter,and 10%separate gel were the suitable settings of the 2-DE protocol.The experiment preliminary established the 2-DE protocols of Fokienia hodginsii leaves which would be beneficial for the study of proteomeAnalysis the differential protein,fokienia hodginsii under the condition of different concentrations of salt stress protein abundance changes are different.Through the analysis software we found significant difference of protein under salt stress,sixteen protein is raise expression and five protein is lower expression.Each protein sample above 3 times repeated experiments,to remove electrophoresis error happened in the process.In the difference of amount expressing,most of the protein concentrated on the acidic side from pH4 to pH5 and located in the middle of acrylamide gel.The protein distributed dispersively in the five points of decrease expression.From the mass spectrometry of 21 protein with technology of two-dimensional electrophoresis,20 protein has been tested successfcully and found the number after searched with Mascot.Input the number in protein website to search the name and function query the corresponding protein in which the analysis result has shown that the function of differential protein mainly reflected in the next several aspects.Carbon metabolism have 3 relative protein;Glyoxylate and dicarboxylate metabolism have 5relative protein.Carbon fixation in photosynthetic organisms have 3 relative protein;Citrate cycle(TCA cycle)have 2 relative protein;Phenylpropanold biosynthesis have 2 relative protein;Oxidative phosphorylation have 2 relative protein;cysteine and methionine metabolism have 2 relative protein.