The Isolation And Identification Of Endophytic Fungi From Dendrobium Sinense And Host Trees

Orchidaceae is a typical mycorrhizal plant,and most species have a high ornamental and medicinal value.Because of such a precious,it triggered a man-made excessive excavation and ecological damage.Under the natural environment,the seed germination and seedling growth of Orchidaceae must be endogenous fungi.According to the previous study of host on the host trees have a preference,its many factors,such as nutrition,plant hormones,fungi and so on.In this study,we investigated the presence of fungi in orchids and host trees fungi from the perspective of fungi.In order to explore the correlation between endophytic fungi between D.sinense and host trees,the fungi were isolated and purified by the individual and root system of D.sinense and the corresponding host trees.By combining traditional morphological techniques and molecular biology identification techniques,To study the relationship between Orchidaceae and endophytic fungi of host,so as to screen and identify the endophytic fungi which had important effects on D.sinense,and isolated fungi from root and shoot of D.sinense.The distribution of fungi was analyzed according to different seasons.The isolation and identification of samples collected in dry season and rainy season were studied by tissue isolation method and grinding embedding method.The main results are as follows:?1?Isolation results:351 fungi were isolated from the dry season,and 147 strains were obtained after purification.The average isolation rate of endophytic fungi was 57.0%,the nutrient roots?G?,protocorms?Y?,roots and leaves?SG,SJ,SL?of adult plants were 57.0%)were 40.4%,20.5%,76.4%,76.7%and 70.9%,respectively.The average isolation rate of host trees endophytic fungi was 89.1%,and the host trees?GP?reached100.0%,seedling host trees?SP?was 89.5%,and protocorm host trees?YP,78%?was lower.The 385 strains of fungi were isolated from the rainy season and 144 strains were purified.The average isolation rate of endophytic fungi was 81.2%,the vegetative roots ?G₂?,the root and shoot?SG₂,SJ₂,SL₂?of the seedlings were 59.3%,respectively,and the isolation rate of the sampling materials was 80.4%,80.5%,91.1%and 94.1% respectively.The average isolation rate of endophytic fungi was 90.7%,the root of vegetative root?GP₂?was 93.1%,the seedling host trees?SP₂?was 88.2%The maximum reproducibility of D.sinense was 13.0%higher than that of host trees?12.9%?,and the maximum reproducibility of D.sinense was 20.0%and 22.5%,respectively.The results of the sampling of D.sinense and the host trees were found to be better than D.sinense,but the diversity of the two species was more abundant than that of the host trees.?2?Morphological identification.The morphological observation showed that there were 7 families and 8 genera of the genus Dendrobium in the dry season.There were 3 enera and 3 genera in the host endophytic fungi,among which the genera belonged to Aspergillaceae?Aspergillus sp.?,Xylariaceae?Daldinia sp.and Nemania sp.?,and Trichoderma sp.of the Hypocreaceae,in which the carbon sponge Nemania and richoderma were the dominant genus.In the rainy season,the genus D.sinense was identified as 4 families and 5 genera,and the host trees was 4 families and 4 genera.Among them,they belong to the genus Nectriaceae?Fusarium sp.?,Xylariaceae Nemania sp.),Hypocreaceae?Trichoderma sp.?and Cladosporiaceae?Cladosporium sp.?.The dominant genus is the other three genera except Cladosporium sp.The results howed that the common genera of D.sinense and the host trees were Nemania and Trichoderma.?3?Molecular biology identification.The results of fungi identification of .sinense and host trees in the dry season were 24 species and 31 genera,which all the ungi identified in the dry season were 17 families and 29 genera,the rainy season were 15families and 21 genera.The endophytic fungi of D.sinense were identified in 15 families and 21 genera,and the endophytic fungi were 14 families.The endophytic fungi and their hosts were 11 genera and 37.9%,of which 7 were dominant genus.The endophytic fungi of D.sinense were identified in 13 families and 18 genera,and their host identification was9 families and 12 genera.The similarity was 42.9%of D.sinense and the host trees,and 6genera were dominant genera.Molecular biology identification was more accurate than morphological identification.?4?Correlation analysis.The total of 31 genera of D.sinense and host fungi in the dry season was identified,including 29 genera of D.sinense and 21 genera of the host trees.There were 16 genera between the two genera,while the host trees had five unique,which found that the distribution of strains of Dendrobium was more abundant than the host. mong the 16 common genera,Xylaria,Colletotrichum,Pestalotiopsis,Trichoderma,Nemania,Fusarium of the six genera were dominant.At the same time,through the statistical analysis of the community results at the classification level,it was further confirmed that the six fungi were distributed in D.sinense and host trees.It was inferred that these fungi have play a"bridge"role in D.sinense and the host trees...