The Differentiation Of Protein Expression Of Ms2 In Taigu Nuclear Male-sterility Wheat

Use Taigu sterile near-isogenic lines of wheat (Lumai 15, Ms2 Lumai 15 and Rht10 + Ms2 Lumai 15) as the materials, and material parts are panicle in different periods. Using of 2-DE technology and mass spectrometry, differentially expressed protein of Ms2 were studied, The main results were as follows:1. The two-dimensional electrophoresis fitting to the young panicle protein of common wheat has been set up through many repeats.Precipitation protein with TCA-acetone method, tested the total proteins of meiosis, uninucleate and binucleus stage of the three materials and achieved good results. Detected with PH4-7,24cm prefabricated dry linear strips for IEF of the extracted proteins,then separation with the concentration of 12% polyacrylamide gel electrophoresis and gain good results.2. In the pH4-7,10-115KD,there are about 500 protein spots at meiosis stage, 600 spots at uninucleate stage and binucleus,respectively. Most of the protein in the pH4.5-7(strip alkaline side),10-80KD. The differentiation of the protein expression between the immature ears of experimental lines has been found.3.The peptite fingerprinting(MALDI-TOF-MS) of 80 protein spots of the three phases were analyzed by matrix-assisted laser desorption time of flight mass spectrometry and used Mascot to search in the swissprot datebase,which identifyed 47 protein spots that had gotten scores above 50.The last 30 protein spots which gain lower scores (less than 50 points) were high performance liquid chromatography-mass spectrometry (HPLC-MS/MS) and 8 protein spots were identified and obtained better peptide fingerprinting.4. Elastase inhibitor 4 and 50S ribosomal protein L6 were identified on meiosis, TPR repeat-containing thioredoxin TTL1,ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit and mitochondrial ATP synthase precursor were identified at the uninucleate stage; Phosphoglycerate kinase,quinone reductase 2 and J-domain protein were identified at the binucleate stage. In addition, some proteins may be related to fertility: at the meiosis stage, Chalcone--flavonone isomerase, cyclin-dependent inhibitor 1; the F-box protein ORE9, UDP-glucose 4-epimerase , Calcineurin B-like protein 3, GTP-binding protein yptV2 at the stage of binucleus;recombination protein SPO11-2, S-adenosylmethionine synthetase, ras-related protein Rab7 at the binuciate stage.5. Functional analysis known protein,speculated that male sterility of wheat may be related to energy metabolism disorder,signal transduction blocked,accumulation of reactive oxygen species,et. These physiological phenomena are likely to lead to the occurrence of infertility. These differential proteins may directly or indirectly involved in the certain key ways of the fertility and abortion of the tested materials. Protein with carbohydrate metabolism, and ABA synthesis and catabolism-related proteins may directly or indirectly slow the growth of the young spike of wheat.