Study On Culture, Biological Characteristics And Phagocytosis To Bacteria Of Plasmodia In Major Species Of Myxomycetes

Myxomycetes, both having the characteristics of protoza and fungi, are the special and important groups in the eukaryotes. Its natural products have the effect of anti tumor, cytotoxicity and bacteriostasis. Because of its special characteristics, making it the status of the biological world has not been uniform. Meanwhile, due to its disputed status, it provides various methods for research. In this paper, many culture methods have been used to study on biology and activity of Myxomycetes.Agar culture method by different media and nutrient solution has been used to study the optimum culture condition of spore germination and plasmodium growing of Physarum melleum, Physarum globuliferum, Physarum pusillum, Hemitrichia clavata, Hemitrichia serpula, Stemonitopsis hyperopta, Arcyria denudata, Arcyria incarnata, Arcyria cinerea, Badhamia affinis, Cribraria violacea, Diderma globosum.Oats-agar medium and pine needle nutrient solution are the optimum combination for spore germination of Physarum melleum, Physarum globuliferum and Physarum pusillum. Pine needle medium and pine needle nutrient solution are the optimum combination for spore germination of Hemitrichia clavata, Arcyria incarnata, Arcyria cinerea and Diderma globosum. Agaric-agar medium and agaric nutrient solution are the optimum combination for spore germination of Hemitrichia serpula. Mircoelements-agar medium and pine needle nutrient solution are the optimum combination for spore germination of Arcyria denudata.The results show that the optimum media and proportion for plasmodium growing of Physarum melleum, Physarum globuliferum and Physarum pusillum is agaric-agar medium, of Hemitrichia clavata, Hemitrichia serpula and Arcyria denudata is miroelements-agar medium, of Arcyria incarnata and Arcyria cinerea are all chosed media except pine needle-agar medium, of Diderma globosum are miroelements-agar medium and oats-agar medium.Moist chamber culture method, oat-agar culture method were used to study the process of the individual development of Physarum pusillum. The life cycle of spore-to-spore was completed on oat-agar culture media. The results showed its complete life cycle including spore germination, a unicellular amoeboid or swarm cell stage, a multinucleate plasmodium stage and maturity of fructifications stage.Liquid fermentation combined with oat-agar culture were used to produce plasmodium of Physarum.Then plasmodium was induced to obtain fruitification in lab by hunger and adjusting light (3000,6000,9000,12000lx) and temperature (20,22,24,26℃). Fruitification of Physarum melleum, P. globuliferum, P. compressum and sclerotium of P. melleum were obtained. On oat-agar medium, the optimum conditions of plasmodium of P. melleum to fruitification were 26℃,6000 lx; of P. globuliferum were 24℃,6000 lx; of P. compressum were 26℃,6000 lx. In liquid medium, the optimum conditions of plasmodium of P. globuliferum to fruitification were 24℃,9000 lx; of P. compressum to fruitification were 20℃,6000 lx. The optimum conditions of plasmodium of P. melleum to sclerotium were 22℃,6000 lx.Cultured plasmodia of 8 Physarales species (Physarum melleum, P. globuliferum, P. compressum, P. cinereum, P. pusillum, P. bivalve, Badhamia cinerascens, and Didymium nigripes) were obtained by liquid methods, determined the suitable media and culture conditions of each species by single factor test. The suitable media was oat liquid media and the optimum conditions of P. melleum were pH 9.0,26.5℃, and 110r/min; of P. globuliferum were pH 9.0, 26.0℃and 100r/min; of P. compressum were pH 4.0,25.5℃and 100r/min; of P. cinereum were pH 8.0,25.5℃and 100r/min; of P. pusillum were pH 5.0,26.5℃and 110r/min; of P. bivalve were pH 6.0,25.5℃and 90r/min; of B. cinerascens were pH 7.0,26.0℃and 100r/min; of Didymium nigripes were pH 9.0,25.0℃and 110r/min, respectively. The results showed the suitable media and culture conditions of Physarales myxomycetes providing the foundation for liquid fermentation of myxomycetes plasmodia.Extracting the sporophores of Physarum melleum by methyl alcohol, acetone, chloroform and petroleum ether, bacteriostasis study has been done using above four extractions by filter paper method. The results showed that four extractions had no bacteriostat activity; however it is the first study on the chemical composition of Physarum melleum.Owing to its ability of phagocytose nutrient substance(including bacterium), the study on the phagocytosis of Physarum melleum, Physarum globuliferum, Physarum compressum, Physarum pusillum, Physarum bivalve, Badhamia cinerascens, Didymium nigripes to had been done. The results showed that the plasmodium and bacterium are symbiosis, the form of phagocytosis is different by different plasmodia, it is concluded that there is bacteriostatic enzyme in plasmodia.