A Study On Diversity Of Endophytic Fungi From Nothapodytes Pittosporoides In Guizhou Province

Endophytic fungi are important fungal resources,which play an important role in the process of ecosystem cycle.Nothapodytes pittosporoides is a kind of important medicinal plant,which contains the important medicinal ingredient camptothecin.This research aims to collect samples of N.pittosporoides from Xiuwen country in four seasons,Wanshan district and Bozhou district of Guizhou province.Separate and purify of endophytic fungi from different tissue parts of N.pittosporoide,and to evaluate its fungal diversity.1037 strains from 1444 N.pittosporoides tissue blocks were isolated,which belong to 3 classes,9 orders,31 genera in Ascomycetes.Among them,656 strains were isolated from Xiuwen,of which,151 strains were isolated from from the spring tissue,114 stains were isolated from the summer tissue,247 strains were isolated from the autumn tissue,144 strains were isolated from the winter tissue;223 strains were isolated from Wanshan,and 258 strains from Bozhou.Based on phylogenetic analysis of ITS and 5.8S rDNA sequences of these morphotypes,the results showed that Dothideomycetes were accounting for 58.1%,Sordariomycetes for 32.3%and Eurotiomycetes for 9.68%.The dominant species was Diaporthe,which had the absolute superiority of 679 strains.Meanwhile Colletotrichum and Phyllosticta of Cladosporium were the second groups with relatively low proportion.Amongst the 31 genera of endophytic fungi isolated,the sporulation and spore structure of 12 genera were observed,of which the sporulation and spore structure of Diaporthe,Colletotrichum,Alternaria and Fusarium were clearer.In addition,12 endophytic fungal genera were identified via both morphology and molecular biology method.They were Bipolaris,Periconia,Zasmidium,Phyllosticta,Hypoxylon,Cercophora,Australiasca,Penicillium,Aspergillus.The distribution of endophytic fungi was different in different seasons.The endophytic fungi quantity in autumn was the highest,of which the diversity index and the Simpson index was higher than those of the other three seasons.In different districts,Xiuwen’s colonization rate was the highest,but Wanshan‘s separation rate was the highest.The three districts had their own unique species respectively,there were 7 in Xiuwen and Wanshan,2 in Bozhou.Xiuwen had the highest diversity index,Simpson index and evenness index for endophytiv fungi,while Wanshan had the highest abundance.In addition,the colonization rate and isolation rate of stem were higher than those of the leaf,while the diversity index,Simpson index and richness index of leaf were significantly higher than those of stem and other tissues.Moreover,with certain host tissues pecificity,the specific tissue of N.pittosporoides had their unique specific of endophytic fungi.