Endophytic Microbes In Dendrobium Catenatum From Three Cultivation Modes

In this study,we collected roots、stems、leaves and environmental substrate samples from 3 different cultivated modes(potted cultivation,cave-living cultivation,living-tree cultivation)in Dendrobium catenatum C15 species.We test the microbe community composition by tissue culturing method and molecular techniques of 16SrDNA,ITS sequencing method.The study on endopyhtic diversity and the relationship between endophyte and medicinal ingredient content accumulation were meaningful to improve resistance,quality,production of D.catenatum by using endophytes The main results were showed as follw:1、Abundant endophytic fungi from D.catenatum C15 species were identified,and the community composition of different culture modes or tissues are different.By 16SrDNA sequencing and ITS sequencing,the bacteria and fungi in the leaves,stems,roots of D.catenatum and cultivated substrate samples were sequenced accurately.The abundance and difference of the endophyte were obtained by OTU cluster analysis.Alpha diversityof analysis were used to measure endophytic microbes in D.catenatum.The abundance of endophyte in the community ecology was measured as well as the trend of species change was obtained.Beta diversity of endophytic species diversity among different environmental communities was studied.By annotated with the RDP database and the Unite database,the species annotation of all OTU in each sample was obtained.The species of endophyte related to medicinal ingredients of D.catenatum was obtained by analyzing the correlation between the endophytes and the medicinal ingredients of D.catenatum.2、A total of 1865 fungal strains were isolated from all D.catenatum samples,belonging to 1 phylum,3 classes,9 orders,17families and 23 genus.A total of 397 fungi strains were isolated from potted cultivation,791 strains were isolated from cave-living cultivation,677 strains were isolated from living-tree cultivation.A total of 103 endophytic fungi were identified by morphological characteristics.According to the molecular biology,47 fungal taxa were identified,belonging to the fungi 1 phylum,3 classes,9 orders,17 families and 23genera,all of these taxa belonged to Ascomycota。3、A total 5 fungi could be co-cultured with tissue culture plant of D.catenatum by Preliminary selection:103 endophyte fungi were co-cultured with host plant and the symbiotic situation were observed per 7 days.Most tissue culture seedlings showed different degree of pathological changes such as swallowed by mycelium,albefaction and yellow,,etc after 60 days.5 strains passed our tests,their number were C-29(Cytospora sacculus)、C-63(Pezicula neocinnamomea)、C-64(Phyllostict capitalensis)、C-70(Nigrospora chinensis)、C-85(Arthrinium phaeospermum)respectively.