Study Of The Proteomes Of Taraxacum Kok-saghyz Rodin In Response To Jasmonate

?Object?Using one proteomics method two-dimensional electrophoresis?2-DE?to establish a two-dimensional electrophoresis system for Taraxacum kok-saghyz Rodin.Study the inducement characteristics of the proteins in Taraxacum kok-saghyz Rodin?TKS?after MeJA treated.Establish the foundation for increasing the amount of gum production by treating the TKS with MeJA.?Methods?By optimizing the selection of sample preparation,the definition of protein quantity,the dyeing method and the pH range of strip,establish an appropriate system for TKS two-dimensional electrophoresis.The study selected three-month old TKS plants leaves for materials,then sprayed 0.4 mmol·L^-1 MeJA on the leaves surface,after 0 and 24 hours,extract leaf protein for two-dimensional gel electrophoresis,then chose the differentially expressed proteins to do mass spectrometry and analyze their function.?Results??1?By comparing the protein concentration and protein extraction rate of five different methods,Tris-HCl extraction method,phenol-ammonium acetate precipitation method and modification were selected for 2-DE electrophoresis.And after analysised the2-DE electrophoresis results of this three methods,the TCA-acetone method was selected for further 2-DE electrophoresis.?2?By analysing the amount of protein loaded,the methods of dyeing and the best conditions of strip,descided to use 1200?g protein sample,17cm?linear,pH3-10?strip and Coomassie brilliant blue as the best condition for 2-DE electrophoresis.In this way the most protein spot can be obtained,and the Coomassie brilliant blue is also compatible with mass spectrometry analysis.?3?Make a gap anlysis for TKS proteomics databases after MeJA treatment by using the optimized 2-DE electrophoresis system.The results showed that there were 20 differential protein spots,and 15 of them are up-regulated,5spots are down-regulated.The 10 spots were chosen for mass spectrometry analysis.The result showed that most of them are related to plant photosynthesis,such as FBAase and RbcS.?Conclusions?The research established an appropriate sample extraction method and a suitable two-dimensional electrophoresis system for TKS leaf proteomic analysis.And then analyze the TKS proteomics databases after MeJA treatment by using the system we established before.And a further mass spectrometry analysis was be used to identify the differential protein spots.This study provide a new way for TKS MeJA-respond research and set a foundation for TKS rational exploitation and utilization.