The Preliminnary Study Of Proteomics To The Resistance Of The Tomato Brown Spot

Tomato (Lycopersicon esculentum Mill) originating in the vicinity of South America of Peru, Ecuador and Bolivia, Annual herb of the genus in the Solanaceae Tomato. Its fruit is nutritious, good taste, bright color, delicious, extensive cultivation, high yield, loved by people around the world, its market demand. According to FAO statistics, from1994to2006, the global tomato export trade volume increase from＄41billion to＄80billion,while the tomato production in China ranks first in the world. But the increase in demand along with the tomato, tomato protected even the kinds of diseases that spread, resulting in the tomato cuts or even never produced a huge economic losses to the tomato industry.Tomato brown spot disease outbreaks in recent years in several regions, pose a serious threat to tomato production, the disease is more common in many parts of the province also reported any outbreaks of disease, tomato production cuts will lead to even never produced in Henan, Guangxi, Zhejiang and Jiangsu dishes District. In many regions of Hei Longjiang province also tended to be worse and spread the trend. Currently, tomato brown spot disease resistance mechanism is not yet very clear, there is no reports of resistance genes, which largely affects the control of tomato brown spot disease. Therefore, the study of tomato brown spot disease resistance mechanism is important. The arrival of the "post-genomic era", functional genomics become the focus of the study, proteomics is its mainstay. Now, there is on proteomics system and a comprehensive study of tomato molecular mechanisms of this disease response, disease resistance-related protein regulatory network research.. Therefore, the use of two-dimensional electrophoresis resistant and susceptible varieties of tomato Differential proteomic research, to explore the protein regulatory networks, it is urgent, far-reaching significance.In this experiment, the first work is screening the tomato disease resistance germplasm resources,used material of50tomato varieties provided by the Tomato Research Institute of the Northeast Agricultural University, Artificial inoculation of different tomato varieties, seedling inoculation.Combination of identification methods are:mechanical inoculation method, inoculation of seedling age of four true leaves, tomato brown spot disease spore concentration is1×107spores/mL. Screened for the results:two resistant varieties,09605and09009;14milddle resistant materials;28susceptible varieties,09011and so on; six very susceptible varieties:.In the two-dimensional electrophoresis study, part of the optimization, including the number of sample crushing, grinding time and the amount of protein samples on the sample, The results showed that more fully in a mortar grinding, grinding time for10min, the protein sample concentration is more suitable for subsequent analysis, sample volume for the protein spots1200ug gel map. Two-dimensional electrophoresis technology, select the tomato resistance、susceptible varieties09605and09011as test material, under the stress of tomato brown spot disease difference proteomics. After repeated trials, a more mature tomato leaf protein two-dimensional electrophoresis system had obtained, proteome changes of tomato resistance to susceptible varieties under tomato brown spot disease stress, after test analysis, the final33of the differentially expressed proteins, Of which09605(R) after inoculation compared with the control, the expression of eight point protein increase, nine point protein expression levels down, new protein of one point, two point of disappearance of protein;09011(S) after inoculation, compared with the control expression were up-regulated five protein spots and protein expression levels down point ten. Peptide mass fingerprinting analysis on these points, successfully identified17protein spots, the PMF data, combined with bioinformatics methods to analyze the specific functions of each protein spot, from the overall level of the resistance mechanism of tomato brown spot disease, is expected to provide theoretical support for the tomato breeding for disease resistance, and to lay the foundation for further study.