Study Of Main Species Of Myxomycetes On Ontogeny, Molecular Systematics And Chemical Component

Myxomycete was an unusual group which both like Fungus and animal kingdom. Because of theirunique biological characteristics, they were used as laboratory materials by Cell Technology and GeneticTechnology. Nowadays, in order to find new medical resources, people were interested in the study ofMyxomycetes on active component. But the utilization of Myxomycetes was constrained because thecultivation and identification were uncertain radically. In order to develop new crude medical resources andimprove the utilized space of Myxomycetes, the yielded fructifications were collected, cultured and studiedon ontogeny by agar culture. Their systematic relationship were analyzed in molecular level. The chemicalcomponent was seperated and identified. This study is the basis of the utilization of Myxomycetes.The specimens and samples were collected in Lu Shuihe, Linjiang, Antu, Jiangyuan and Changchunin Jilin Province. Fructifications of Physarum tenerum, Physarum citrinum, Diachea bulbillosa,Metatrichia vesparium, Arcyria globosa, Physarum viride, Hemitrichia clavata, Arcyria cinerea,Comatricha tenerrima, Stemonitis flavogenita, Comatricha elegans, Trichia erecta, Stemonitis lingnicola,Stemonitis fusca, Ceratiomyxa fruticulosa, Comatricha solitaria, Perichaena depressa and Hemitrichiaclavata were harvested from the moist chamber culture by the substrates collected from those places.Among them, the process from plasmodia to mature fructifications of Physarum tenerum, Physarumcitrinum, Diachea bulbillosa, Metatrichia vesparium, Areyria globosa and Physarum viride were observed.The process from young fructifications to mature fructifications of Hemitrichia clavata and Arcyria cinemawere also observed. Those nineteen species of Myxomycetes were all mature, and their morphologicalcharacteristics were the same as the yielded fructifications. The environment of moist chamber culture issimilar to the living environment of Myxomycetes. Myxomycetes lived well in changed temperature. Moistchamber culture is provided to aid to field collection, and it can make materials for the later research.The differences between the fructifications harvested from moist chamber culture and fieldspecimens were morphologically compared in peridia, spores and capillitia by anatomical lens and lightmicroscopy. The living process of Myxomycetes were described. The results show that the living stage ofMyxomycetes' fructification has its own specification, but there only have some in common.The life cycle of three Myxomycetes in Physarales were observed by moist chamber culture,hanging drop culture and oat-agar culture which include spores, swarm cells, myxamoebae, plasmodia andfructifications. This is the basis for the study of Myxomycetes' specification. The laboratory cultivation methods of Myxomycetes were explored. This study also try to culture other Myxomycetes in Trichiales,but just obtained the plasmodia and the mature fructifications were not obtained eventually. The resultsshow that the Trichiales were difficult to culture compared with Physarales.From all the stages of life cycle, the plasmodia of Physarella oblonga, Physarum globuliferum andPhysarum tenerum are all phaneroplasmodium. The fructifications changed from a series of color transitionand matured eventually. It can be concluded that the life cycles of Myxomycetes in the same Family havesome same characteristics, but every Myxomycete has its' own specification at the same time. For example,the sporation of Physarella oblonga and Physarum tenerum are all opening method, but the sporation ofPhysarum globuliferum is pore method. The color of plasmodia of different species is different. Theprocess of the emergence of fructification has its' own specification. It was the first time to report the lifecycle of Physarum globuliferum and Physarum tenerum.The phylogenetic taxonomy of Myxomycetes were studied by modern molecular biology technique.It was the first time to extract DNA of Myxomycetes by Fungus Genomic DNA Extraction Kit. DNA ofMyxomycetes were also extracted by CTAB method. The results show that DNA obtained by the above twomethods both can be used for molecular biology research. Polymerase chain reaction (PCR) amplificationof rDNA from Diderma lucidum, Tubifera ferruginosa, Physarum globuliferum, Arcyria cinerea,Physarella oblonga and Trichia erecta using three pairs of primers. The PCR products were cloned andsequenced. Four sequences of Arcyria cinerea (ITS rDNA), Physarum globuliferum (ITS rDNA), Didermalucidum (12S rDNA) and Tubiferaferruginosa (12S rDNA) were eventually sequenced. The sequences ofArcyria cinerea (ITS rDNA) and Tubifera ferruginosa (12S rDNA) were submitted to GenBank, and theaccession numbers are EF513148 and EF513149 separately. The four sequences are all the first reports ofthose Myxomycetes' sequences in the world.The chemical component of Trichia decipiens was preliminary studied. The fructifications of Trichiadecipiens were used to extract by methanol. After that, a compound was obtained and purified bypreparation silica gel plate and HPLC method. It is a aliphatic hydrocarbon from the analysis of ~1H-NMR,UV, HPLC and MS.