| Hoya R.Br.is a creeping perennial herbaceous evergreen plant of the Asclepiadaceae family,which has high ornamental value and medicinal value.C hina is one of the main distribution areas of Hoya,which is mainly distributed in the tropical a nd subtropical regions of South of C hina.The current research on Hoya more concentrated in tissue culture,pharmaceutical value and taxonomic research etc.,but there are no reports on diversity and phylogeny of endophytic fungi in different Hoya species.In this study,we used both traditional tissue segments culture method and molecular biological methods to analyze and compare the community structures,diversity and molecular phylogeny of endophytic fungi with leaves of Hoya sp.1 and Hoya sp.2?Hoya sp.1 was collected from C hangjiang County and Ledong County in Hainan island,and Hoya sp.2 was collected from Haikou C ity and Ledong County in Hainan island?Their codes are BWA?JFA?HYS and JFS.The major results and conclusions are listed be lo w:(1)The research use of traditional tissue segments culture method received a total of 205 strains of endophytic fungi were isolated,which were divided into 31 operational taxonomic units.Among them,60 strains of endophytic fungi were isolated from BWA,divided into 14 operational taxonomic units,and classified into Sordariomycetes(71.67%)?Dothideomycetes(26.67%)and Agaricomycetes(1.67%)of Ascomycota(98.33%)and Basidiomycota(1.67%),however,1 operational taxonomic units can only be identified as Sordariomycetes class,the others were classified into 9 genera,8 families,7 orders;42 strains of endophytic fungi were isolated from HYS,divided into 7 operational taxonomic units,and classified into 2 genera,2 families,2 orders,Sordariomycetes,Ascomycota;45 strains of endophytic fungi were isolated from JFA,divided into 11 operational taxonomic units,and classified into 8 genera,8 families,5 orders,Sordariomycetes(77.78%)and Dothideomycetes(22.22%)of Ascomycota;58 strains of endophytic fungi were isolated from JFS,divided into 8 operational taxonomic units,and classified into 5 genera,5 families,5 orders,Sordariomycetes(74.14%)and Dothideomycetes(25.86%)of Ascomycota.(2)The research use of molecular clone detection method received a total of 121 operational taxonomic units among 323 endophytic fungi cloning sequences from different ITS gene clone libraries.Among them,a total of 27 operational taxonomic units among 78 endophytic fungi cloning sequences were received from BWA,and classified into Ascomycota(91.02%)and Basidiomycota(8.98%),however,5 operational taxonomic units can only be identified as class or phylum,the others were classified into11 orders,7classes;a total of 35 operational taxonomic units among 94 endophytic fungi cloning sequences were received from HYS,and classified into Ascomycota(98.94%)and Basidiomycota(1.06%),however,3 operational taxonomic units can only be identified as class,the others were classified into 5 orders,4 classes;a total of 23 operational taxonomic units among 68 endophytic fungi cloning sequences were received from JFA,and classified into Ascomycota(98.53%)and Basidiomycota(1.47%),however,5 operational taxonomic units can only be identified as class or phylum,the others were classified into 7 orders,4 classes;a total of 48 operational taxonomic units among 83 endophytic fungi cloning sequences were received from JFS,and classified into Ascomycota(79.52%)and Basidiomycota(20.48%),however,22 operational taxonomic units can only be identified as class or phylum,the others were classified into 12 orders,8 classes?(3)The diversity index of endophytic fungi was obtained by the traditional method showed that BWA was higher than JFA,HYS was higher than JFS,JFA was higher than JFS,but the results obtained by molecular cloning method were just the opposite;The diversity index of endophytic fungi was obtained by the traditional method was lower than that of molecular cloning method;There were significant differences between the two methods in the detection of endophytic fungi,and some of them could not be detected in the two methods simultaneously.(4)Based on the comparison of the morphological characteristics of fungi,the phylogenetic analysis of ITS gene fragments and the literature review,it is very possible to determine the fungus(BWAJL49)is most likely to be a new species of Sordariomycetes,and two fungi represented by JFSJL16 are likely to be Endomelanconiopsis microspora,which is a newly recorded species in C hina.The results are helpful to form a more comprehensive understanding of the community structures and diversity of endophytic fungi in Hoya R.Br.,and provide theoretical basis and resource basis for the understanding and utilization of this important resource,has theoretical and practical significance. |
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