Study On Artificial Mvcorrhizal Synthesis And Shiroes Soil Microbial Diversity Of Thelephora Ganbajun

Thelephora ganbajun Zang is a wild,endemic edible mushroom in Yunnan,belonging to the genus Thelephora in the family Thelephoraceae and phylum Basidiomycota.It forms symbiotic association with a variety of Pinaceae species,as a kind of ectomycorrhizal fungus.It is impossible to produce mature fruiting bodies of T.ganbajun on artificial media,like the other saprobic edible mushrooms.Thus,only the limited wild growing mushroom which makes it highly commercial value.The endangered situation and ecological degradation,caused by over-picking has seriously hampered the development of T.ganbajun industry.In this thesis,the comparative study on the shiroes situation in the forming period of wild T,ganbajun,Boletus spp.and Rhizopogon spp.was carried out,providing the morphological basis for the formation of T.ganbajun fruiting body and mycorrhizae;In order to explore the formation of T.ganbajun mycorrhizae,the artificial infection experiment was carried out using T.ganbajun hyphae with the seedlings of Pinus kesiya var.langbianensis and Pinus yunnanensis Faranch;Soil samples of T.ganbajun shiroes and non-shiroes were collected from four localities in Yunnan Province.Bacteria and fungi were cultured by traditional plate culturing method,and the DNA sequences of ITS and the V3,V4 regions of 16S rDNA in soil samples of shiroes and non-shiroes were sequenced useing Illumina Miseq high-throughput sequencing technique,to compare the diversity,similarities and differences of culture-independent fungal and bacterial communities in the two soil samples,which will provide theoretical support for artificial culture of edible fungi,including T.ganbajun,Boletus edulis,Tricholoma matsutake.The main results were as follows:1.A large number of mycorrhizae and mycelium were found in the shiroes of T.ganbajun,Boletus spp.and Rhizopogon spp.,there are some differences between the three kinds of ectomycorrhizal structure,which can be used as an auxiliary method to determine and monitor the corresponding fungal species.2.Mycorrhizae of T.ganbajun was successfully synthesized on the seedlings of P.kesiya var.langbianensis and P.yunnanensis Faranch,and the mycorrhizae colonized on the cuttings of P.kesiya var.langbianensis seedlings was obserred to form mantle,and the growing condition and number proved to be the best.3.Using the traditional plate isolation method,53 strains of fungi and 102 strains of bacteria were isolated and purified from the shiroes soil,and 65 strains of fungi and 60 strains of bacteria were isolated and purified from the non-shiroes soil.The analysis of phylogenetic relationship was based on ITS and 16S rDNA gene sequenceing.The results showed that there were.some differences in fungal and bacterial communites between two types of soils,but the predominant fungal categories on genera level were Penicillium,Trichoderma and Mortierella,while Bacillus,Burkholderia and Streptomyces were the dominant bacterial genera in the two types of soils.Through the analysis of high-throughput sequencing data,the results showed that the diversity of bacteria and fungi in the shiroes soil were higher than in the non-shiroes soil,but the difference was not significant,and the microbial composition of the shiroes soil samples in four areas is similar.