Analysis Of Community Structure Of Endophytic Fungi From Several Varieties Of Camellia Sinensis Based On Microbial Culture-Independent And High-Throughput Sequencing Methods

The endophytic fungi of tea tree refer to the fungi that grow in the healthy tissue of tea tree not causing obvious pathological changes.Based on the cultivation conditions and geographical climate of different varieties of tea tree,the endophytic fungi in the tea plants formed a specific community structure and were in rich of microbial diversity.The combination of traditional microbial cultivation technology and modern high-throughput sequencing technology can make the research results more reliable and comprehensive.In this paper,the endophytic fungi from different varieties of tea trees(different varieties of the same geography,the same variety of different geography)were separated,purificated,identificated by molecular biology and the community structure was analyzed.On the other hand,the diversity and community structure of endophytic fungi from Shuchazao of different ages(1～2 years,20 years,40 years and 60 years old)Shucheng County were analyzed comprehensively and systematically through high-throughput sequencing technology.The results are listed as below:1.The separation and purification of endophytic fungi in different tea trees and the analysis of community structure.17 fungal strains were isolated and cultivated from 12 different geographic tea trees.All those endophytic fungi included Colletotrichum sp.,Guignardia mangiferae.,Alternaria sp.,Phomopsis sp.,Penicillium sp.,Aspergillus versicolor,Botryotinia fuckeliana,Pleosporales,Glomerella cingulata,Cladosporium perangustum,Diaporthe eucalyptorum,Pestalotiopsis sp,Ascomycota.sp.,and two unknown strains.Among them,the Colletotrichum sp.(especially Colletotrichum gloeosporioides),Guignardia mangiferae and Alternaria sp.are the most abundant and dominant fungi in these tea plants.The separation rate of endophytic fungus from leaves of middle and lower parts of trees(different varieties of the same geography)are much higher than upper part.And there is unobvious difference of the isolated species between different leaf layers.For the tea trees of the same variety of different geography,the separation rate and the number of isolates of tea trees from Hefei are higher than that of Dongzhi county and Shucheng county.2.The analysis of endophytic fungi diversity from Shuchazao in different ages by high-throughput sequencing methodWe used high-throughput sequencing method to analyze the community structure and the diversity of endophytic fungus from Shuchazao in four ages(1～2 years,20 years,40 years and 60 years)groups to get the group composition of the endogenous fungi in the phylum,the class,the order,the family and the species levels.Among them,the top 10 species at the classification level of the genus were respectively:Saccharomycopsis,Cladosporium,Strelitziana,Pseudoramichloridium,Zymoseptoria,Penicillifer,Trichoderma,Pseudeurotium,Paraconiothyrium,Melanconiella.The fungi of Cladosporium,Strelitziana and Zymoseptoria have an advantage.The abundance and diversity of endophytic fungi in soil were higher than that of leaves,and the number and variety of fungi were different in different tree ages.Among them,there was the highest abundance and diversity of endophytic fungus in the rhizosphere soil of Shuchazao of sixty years old.The abundance of Cladosporium,Strelitziana and Zymoseptoria in the leaves of was significantly higher than that in the rhizosphere soil.The abundance of Strelitziana and the maturity of the leaf layers were positively correlated with the age of the tea tree.The abundance of Zymoseptoria was decreased accompanied by the tea tree grow.The difference of diversity is small when the age of tea tree is almost near,and even when the age is old enough.