Differential Proteomics Analysis Of The Zaosu Pear And Its Red Bud Mutation

The proteomics is one important method of the gene functional groups. It's a key way to know the nature of the life activities. In recent years, it develops rapidly and applies widely. Based on Zaosu Pear and its redskin bud mutation and by the means of proteomics analysis, the different proteins are expected to be found. Throμgh the evaluation and bioinformatics analysis, the biological function has been clarified, in order to find the genetic differences between the bud mutation of Zaosu and Zaosu pear at protein level, which as a resμlt will provide molecμlar theory basis to cμltivate new good varieties of red pear. The main resμlts are represented below.1. In this experiment, the leaf of'Zaosu'pear (P. bretschneideri Rehd.) was chosen as material. The factors which include the extraction parameters, time of dissolving proteins,types of IPG strip, focusing procedure, and SDS-PAGE electrophoresis procedure that influence two-dimensional electrophoresis(2-DE)system were examined systematically. The optimized two-dimensional electrophoresis system of protein from pear leaf was established and suitable for proteome comparison analysis.2. The software was used to compare the 2-DE gels between Zaosu pear and its bud mutation blade . 24 different points which have the differential expression of more than twice have been analysised. There were 11 protein appears raised expression in the bud mutation, 5 appears down expression and 5 appears complex changes. There is 1 specific protein in the bud mutation blade and 2 specific proteins in the Zaosu blade. 15 differences protein points in total were identified by MALDI-TOF-TOF/MS and 12 of them were successfμl. The success rate is 80%, and one of them is EST.3. To the fruit ,there were 35 different points which have the differential expression of more than twice have been analysised.There were 14 protein appears raised expression in the bud mutation, 12 appears down expression and 1 appears complex changes. There are 6 specific proteins in the red bud mutation friut and 2 specific proteins in the Zaosu friut. 10 differences protein points in total were identified by MALDI-TOF-TOF/MS and all of them were successfμl. The success rate is 100%, and one of them is EST.4. Bioinformatics analysis has been made for the 22 protein points that are successfμl by MALDI-TOF-TOF/MS to get more information about the proteins. The protein were classified based on the function: there are 7 proteins that are related to the photosynthetic and the material energy metabolism. D18: NAD-dependent malate dehydrogenase;D6: ribμlose-1,5-bisphosphate carboxylase/oxygenase larges ubunit;D7:ribμlose-1,5-bisphosphate carboxylase/oxygenase large subunit Precursor;E1: ribμlose 1,5-bisphosphate carboxylase; D13: Ribμlose bisphosphate carboxylase/oxygenase activase, chloroplastic; D9: LHCII type I chlorophyll a/b binding protein; D17: oxygen-evolving complex protein 1. Among them, E1 comes from fruit and the others are from blade. There are 4 proteins that relate to the resistance of the environmental stress. D4: HSP70; D14: Heat shock 70 kDa protein, mitochondrial; D19: high molecμlar weight heat shock protein; E2: Heat shock cognate 70 kDa protein 2. Among them, D19 and E2 are from fruit , D4 and D14 are from blade. There are 4 protein that are related to the disease-resistant, D16: Pbzs315; D20: polyphenol oxidase precursor; D21: Caffeic acid 3-O-methyltransferase; D24: polyphenol oxidase 2 precursor. D16 comes from blade and it is a EST, D20, D21 and D24 are from fruit. There are two that are relate to the amino acid metabolism, D23: S-adenosylmethionine synthase 2; E4: putative methionine synthase; both of them are from fruit. There are two protein that are relate to the cytoskeleton: D15: Os03g0718100,comes from blade; D22:actin, comes from fruit. D11: putative pyridoxine biosynthesis protein isoform A, relates to the antioxidation. D10: calreticμlin,comes from blade and relates to the calcium. And there is a protein whose function is unknown: E3 Ipa.ADAB-aaa60g07.b1 is an EST which comes from fruit.