Corynespora cassiicola is a cosmopolitan fungus with a broad host range. One of the hosts often severely impacted is the rubber tree, Hevea brasiliensis on which it causes devastating leaf defoliation. To date, control of the disease is mainly focused on the application of fungicides, but this only provides short-term solution. Functional genomics through transformation allows evaluation of the disease effectors which may eventually lead to more precise control measures to curb the disease. A transformation system for C. cassiicola mediated by Agrobacterium tumefaciens was successfully established with resistance to hygromycin and DsRed as markers. The protocol was optimised with regard to Agrobacterium strains, C. cassiicola isolates, and other parameters. Combination of isolate CLN23 and A. tumefaciens strain LBA4404 carrying pCAMDsRed gave the highest number of transformants. The optimum fungal age, temperature for incubation and co-cultivation period were 7 days old, 28 °C and 4 days, respectively. However, these parameters are highly subjected to interaction between the variables. The transformants carried the DsRed reporter gene, and demonstrated readily apparent red fluorescence, which allowed disease development in planta to be observed. This robust transformation system should pave the way for future genetic manipulations of C. cassiicola.

棒状杆菌Corynespora cassiicola是一种大都会真菌，具有广泛的宿主范围。经常遭受严重破坏的寄主之一是橡胶树巴西橡胶树，它在橡胶树上造成毁灭性的落叶。迄今为止，该病的控制主要集中在杀真菌剂的应用上，但这仅提供了短期解决方案。通过转化的功能基因组学可以评估疾病的影响因子，最终可能会导致更精确的控制措施来遏制疾病。成功建立了以根癌农杆菌和DsRed为标记的根癌农杆菌介导的C. cassiicola转化体系。该协议针对农杆菌菌株进行了优化，C. cassiicola分离株和其他参数。分离株CLN23和携带pCAMDsRed的根癌农杆菌LBA4404的组合得到最高数量的转化体。最佳真菌年龄，孵育温度和共培养期分别为7天，28°C和4天。但是，这些参数高度受变量之间的相互作用的影响。转化体带有DsRed报告基因，并显示出明显的红色荧光，可以观察到植物体内的疾病发展。这种强大的转化体系应该为未来C. cassiicola的遗传操作铺平道路。