The Proteomics Analysis Of Typical Lesions Of Poplar Canker (Botryosphaeria Dothidea)

Poplar is a very important economic species. Poplar canker disease is one of the major diseases which damage poplar trunks and branches. It has significant economic and ecological significance that study the pathogenic mechanism of poplar canker disease. Proteomics is an emerging research area after the genome era, it is an important means to reveal the physical phenomenon of life in today’s biological research, it is an important method that reval the life of tree biology. The proteomics analysis of typical lesion of poplar canker disease is very helpful for elucidating the pathogenic mechanism and the resistance mechanism of poplar canker. It also provides reference for the proteomics study of other trees interaction with pathogen. Bark of107poplar which was collected in September was used as the experimental material. A suitable method for extracting the protein of poplar bark was established. And the proteomic method was used to screeing the differentially expressed proteins of the typical lesion which was produced by the interaction of poplar and canker pathogens, Differential expressed proteins were identificated by mass spectrometry and bioinformatics analysis. The main results were showed in this research are as following:1. A kind of two-dimensional electrophoresis method that suitable for poplar bark is established. For the protein extracting process, the settling time of the sample and the volume of lysis solution and the lysis time of different gradient were setted. The optimal conditions of extracting the protein of poplar bark were established, the optimal (?)recipitating time of the protein was12hours; the optimal amount of the lysis solution which is added to the sample powder was500μL per garm; the optimal lysis time was3(?)uors.In this study, the good effect of the isoelectric focusing (IEF) program is determined hrough many experiments:Grad500V,2hours; Grad1000V,1hours; Grad8000V,(?)0000vh; Step8000V,60000vh. In the reseaech, the IPG strip of pH4-7that with the length of24cm was used, and the concentration of the separate gel is12percent. Good results were gained.2. According to the analysis of the two-dimensional electrophoresis gal map of the (?)ealthy tissue and lesion tissue of poplar bark. A total of54differentially expressed proteins were detected. Of which21protein spots were qualitative differential expressed, and17(?)rotein spots were only detected in the healthy tissue protein gel map; The other5protein pots are only detected in the sick tissue protein gel map. A total of33protein spots were ualitative differentially expressed, take the healthy gel map of the bark protein as a control, a number of20protein spots are up-regulation and the other11proteins spots are down-regulation in expression of the sick tissue protein gel map,3. There were46protein sptos chosen for mass spectrometric analysis, After being analysed by Mascot software, the datebase was used toquery the results, the basic attributes of these differential proteins are identificated, such as the isoelectric and the molecular weight and so on. Take the detect score of the results≥73as the standard, a number of22proteins were tested and were positive, a total of6protein spots were clear identificated, most of the others are predicate protein. These six proteins were:ascorbate peroxidase, heat shock protein, cellulose synthase, leucine rich-repeat receptor-like kinases, bark storage protein and enlase. These proteins are involved in defense response, signal transduction and regulation, the metabolism of amino acid, energy metabolism; transcription and other physiological processes. These proteinss may be associated with the resistance for poplar resistance to canker pathogen.