Proteomic Analysis of Bud Differentiation between Cytoplasmic Male-Sterile Line and Maintainer in Tobacco

Abstract

To explore the molecular genetic mechanisms of cytoplasmic male sterility(CMS) in tobacco,we studied the differential proteins between the CMS line and its maintainer.Immobilized pH gradient two-dimensional gel electrophoresis(2-DE) technique was used to separate the protein spots while gels were stained with the Coomassie Blue G-250.The difference between protein maps of bud from cytoplasmic male-sterile line MSK326 and maintainer line K326 was analyzed with PDQuest image software.Matrix-assisted laser-adsorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS) technique was used to obtain the peptide mass of differentially expressed protein spots.The Mascot software was used to search the protein database NCBInr to identify those spots interested.A total of 365 protein spots were detected within Mr 14.4-97.6 kD and pH 4-7.Seven protein spots appeared in the protein map of maintainer line K326 but absent in that of CMS line MSK326,which were identified as ribulose-1,5-bisphosphate carboxylase/oxygenase,polyphenol oxidase,patatin homolog,and PSI 9 kD protein.It was inferred that the male sterility of MSK326 might be related to energy metabolism turbulence,abnormality in male organ development,deficient nutrition supply and immunity,and inhibition of energy transfer.