Screening Of Pathogenesis Related Proteins Mediated By Rice Disease Resistance Gene Xa21

Rice (Oryza sativa) is a very important crop, rice disease can reduce the yield of rice, it can even result in the lost of harvest. Rice bacterial blight is one of the most important rice diseases.The study to the mechanism of rice disease resistance gene has great significances, The study can benefit us of protecting rice from destroy, enhance rice yield and solve the problem of world people's famine. Xa21, which was cloned by Song et al in 1995, is a resistance gene of rice bacterial blight. Rice bacterial blight was caused by the bacterial pathogen, Xanthomonas oryzae pv. Oryzae, Xoo. We have known that XA21 mainly contains three domains, the Leucine Zipper, an extracelluar leucine-rich-repeat domain, which includes 23 LRR, the transmembrane domain and a serine-threonine kinase-like domain. The characteristic of the receptor-like kinase XA21 has been researched systematically in the latest research in bacteria expression system. To find pathogenesis related proteins of rice disease resistance gene Xa21, the technique of two-dimensional electrophoresis and mass spectrum analysis were used. The experiment establishes the foundation for the research of the interaction of protein and protein and biochemical prosperities analysis.The method that we practiced in detail are as follow. First of all, the rice leaves of trans-genetic material 4021 were inoculated the bacterial pathogen, Xanthomonas oryzae pv. Oryzae, Xoo.After the leaves proteins were extracted by the method of TCA- acetone, the method of two-dimensional electrophoresis was used to separate the proteins. Secondly, we utilize the software to analysis the 2DE maps. The different spots were analysed by MALDI-TOF. 48 differential expression proteins were obtained. Combined to former microarray results, four proteins may relate to the Pathogenesis pathway of Xa21. The E.Coil bacterial expression system is used to clone and express the four proteins, when confirm that the target proteins were expressed abundantly enough, we purify the proteins and prepare antibodies. The experiment establishes the foundation for the research of the further study of the mechanism of disease resistance.