The Proteomics Research Of Virescent Mutant Of Cucumber Leaf

Objective:The experiment was based on both leaves tissues from the first euphylla to the sixth in normal cucumber and virescent mutant, two-dimensional electrophoresis (2-DE) were used to investigate the total proteins of the both materials,in order to detect the different expressed proteins between normal cucumber and virescent mutant,and then identify parts of proteins using the mass spectrometry (MS) technology, and look forward to discovering proteins that may concern with aetiolation. Accordingly, the experiment was settled the foundation of molecule mechanism of the virescent mutant in cucumber.Methods:Both normal and virescent mutant cucumber plants were growed under the same conditions.The leaves from the first euphylla to the sixth were collected when they were present. Following, both the normal and the mutant samples were quick-freezing though the liquid nitrogen, and then preserved at-70℃freezer. The total leaf proteins of the two samples were extracted by TCA-acetone method, after accurately quantitating, analysed by two-dimensional electrophoresis (2-DE) and scanned the gels of good repetitiveness. Subsequently, professional PDQuest software were applied for analyzing and detecting.Different expressed proteins of both materials were found out and then analysis by MALDI-TOF-MS at last.Results and conclusion:The proteomic approach to identify proteins that potentially involved in normal and virescent mutant cucumber leaves were established, and provide the technology platform for the protein research in plants. By the analysis of software,63 proteins exit differences between the normal cucumber and the virescent mutant.22 proteins were isolated and identified using MALDI-TOF mass spectrometry, in which,7 proteins were up-regulated in virescent mutant of cucumber,6 proteins were up-regulated in normal cucumber,others were no obviously different, these proteomics data was the basis of molecule mechanism of etiolation mutant.