The Differential Expressed Protein Profile Research Of Anana Induced By Fussarium Wilt Disease Through ITRAQ Method

Banana fusarium wilt is one of the most critical diseases caused serious damage for banana. Banana fusarium wilt is a fungal disease spread through soil, irrigation water and infected bud materials. If banana infected by Fusarium oxysporum f. sp. Cubense during the growth process, it will happen related pathological reaction, including the physiological and biochemical changes of metabolic pathway, protein expression regulation, gene expression regulation and so on. Lots of studying related to genes differentially expressed, banana fusarium wilt resistance analysis and transcriptome has been done, but the study of protein expression profile about banana fusarium wilt is rarely reported. In this experiment, banana leaves was inoculated with Fusarium oxysporum f. sp. Cubense and samples were get at different time points to study protein expression profile changes using iTRAQ technology, screening and identifying significant differentially expressed proteins related to banana fusarium wilt pathologic reaction. The results were as following:1. The experiment of in vitro inoculation on banana leaves with Fusarium oxysporum f. sp. Cubense was carried out, we sampled pathogenic leaves at different time points for protein extraction and two-dimensional electrophoresis. As a result, the two-dimensional electrophoresis system for banana leaf was build up.2. The iTRAQ technology was used for protein identification and the relative quantitative analysis. We got318539pieces of total spectra, of which matched100559pieces of spectra to bank by Mascot software, we got75635pieces of unique spectra,27563peptides,23426unique peptides. A total of6896banana proteins were identified, of which539showed significantly differentially expressed protein from four time points, including293significantly up-regulated proteins,327significantly down-regulated proteins,37unknown proteins and35unnamed proteins.3. We analyzed the protein expression profile by iTRAQ technology. In4h/CK, there were32up-regulated proteins and31down-regulated proteins, which reached5.4%and5.23%of the total significantly differentially expressed protein; in24h/CK, there were30up-regulated proteins and26down-regulated proteins, accounted for5.06%and4.38%; in6d/CK, there were155up-regulated proteins and210down-regulated proteins, accounted for26.14%and35.41%; in24h/4h there were19up-regulated proteins and21down-regulated proteins, accounted for3.2%and3.54%; in6d/4h, there were174up-regulated proteins and160down-regulated proteins, accounted for29.34%and26.98%; in6d/24h, there were103up-regulated proteins and115down-regulated proteins, accounted for17.37%and19.39%. 4. Compared with CK,4h/CK,24h/CK,6d/CK are together enriched pathway are the biosynthesis of secondary metabolites, metabolic pathway, metabolism of GSH and ribosomes. We analyzed six pathway maps, there were177significantly differentially expressed proteins in the plant-pathogen interaction pathway, which was29.56%of the total significantly differentially expressed protein, there were134significantly differentially expressed proteins in the plant hormone signal transduction pathway, accounting for22.60%.5. From plant-pathogen interaction pathway and plant hormone signal transduction pathway, we found that there were5and8significantly differentially expressed proteins. Pathogenesis-related protein PRB1-3and Mitogen-activated protein kinase12were existed in two pathways above.6. In order to analysis the11proteins which were found in plant-pathogen interaction pathway and plant hormone signal transduction pathway, we found out the corresponding gene sequences for RT-PCR verification, the result showed that the protein expression spectrum data was real and reliable.