Preliminary Proteomic Analysis of the Proteins of Thermo-sensitive Genetic Sterile Rice Anther

The proteins of thermo-sensitive genetic sterile rice anther were separated by two-dimensional electrophoresis with immobilized pH(3-10) gradients as the first dimension and SDS-PAGE as the second.Mean standard deviations of about 1.45±0.23 mm for IPG-IEF and 1615±0.17 mm for SDS-PAGE were obtained.The silver-stained protein spots were analyzed and identified by employing an improved procedure,including (1) in-gel reduction,alkylation and enzymatic digestion;(2) extraction and desalting by using the Ziptip TM;(3)direct MALDI-TOF mass spectrometry analysis and protein database searching.60 protein spots out of about 950 detectable spots on the 2D-gels were identified.The 2D protein maps of the rice anther during the sterile and fertile stages were compared.While the anther changed from sterile to fertile stages,about 20 protein sports showed variations that were significant and reproducible.Five up-regulated proteins and three down-regulated proteins were identified.