A Study On Differently Expressed Proteins Between Ft-10 Mutant And Wild Type Of Arabidopsis

It is well known that FT gene plays an important role in photoperiod pathway regulating flowering in Arabidopsis. Thus the comparative researches between ft mutant and WT might provide a sound basis for better understanding in the molecular mechanisms of plant flowering .Two dimensional electrophoresis and MALDI-TOF MS analysis are used to analyse the differently expressed proteins between the ft-10 mutant and its wild type of Arabidopsis. It is hoped that these experimental results might be helpful to provide information for the molecular mechanisms of plant flowering. The main results show as follows:(1) To optimize two-dimensional polyacrylamide gel electrophoresis(2-DEE) system for the separation and quantification of plant protein species, suitable 2-DEE system was developed by selecting different components and concentrations in isolation solutions and lysis buffers. Sample was prepared by using 0.1M ammonium acetate in methanol(NH4Ac/methyl).The isolated proteins were lysised in lysis buffer consisting of 8M urea,2M Thiourea,4%CHAPS, 65mM DTT ,2%Ampholine and 1mmol/L PMSF. The separated proteins on the 2DE gel were detected by Coomassie brilliant blue R250 staining. It has high-quality and quantity separation of leaf total proteins in this 2-DE image.(2) 34 differentially expressed proteins in the leaf total proteins betweenft-10 mutant and its wild type were digested in gel by trypsin and analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Totally, there are 26 proteins were identified, the success rate is 76%. Among these,16 are up-regulated(spots 3, 5, 6, 10, 11,12, 15, 16, 17, 18, 20,23, 24, 25, 30, 35); 11 are down-regulated (spots 4, 7, 8, 9, 14, 22, 26,28,32,33, 34), and 2 are new synthesis proteins (spots 21,27). According to their functions, these proteins were divided into the following categories: Transcription, Defense, Signal transduction, Energy, Storage protein, Cell growth and so on.