| In this study, two-dimensional electrophoresis and Matrix-assisted laser desorption/ionization-Time of Fingerprinting-Mass Spectrometry (MOLDI-TOF-MS)was used to analyse the proteins from the leaves of chrysanthemum which treated in 4 °C low temperature for 2h,4h,6h,12h,ld,2d and 3d.The chrysanthemum include the chilling-sensitive Dendranthema cv.pekinense(0#) and the chilling-tolerant Dendranthema×morifolium (3#).Some improvements in sample preparation, protein solubilition and so on were made in this study.The interference of rich polysaccharide, lipids and pigments on two-dimensional electrophoresis was solved and an higher repeatability and better separating protein pattern could be gained by this techn ique. At the same time,a efficient solubilition buffer which containing 7M Urea ,2M Triourea, 4 %CHAPS, 5mM DTT has been definded. Stained with Coomassie brilliant blue, the proteins which changed in low temperature stress were analyzed by ImageMaster software 2D Elite. It was detected nearly 500-800 protein spots, After matching,there are 54 differrenr spots in the 0#,52 differrenr spots in the 3#,7 differrenr spots between the 0# and 3#. These differentially expressed proteins were investigated by MALDI-TOF-MS technique. About 63 differentially expressed proteins had the peptide mass fingerprinting(PMF) successfully. The results after Mascot database searching showed that most of the proteins hit putative proteins, and only 12 showed some similarity with known proteins in the database, they are polygalacturonase,glycine-rich RNA binding protein-like,fructose-bisphosphate aldolase ,myb-related transcription factor LBM2, calcium/calmodulin-regulated receptor-like kinase,phosphoglycerate mutase-like protein,cytochrome-C oxidase,ribulose-l,5-bisphosphate carboxylase small subunit seperately. This study will provide clues for the mechanisms of toleranceing low temperature of chrysanthemum.
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