Studies On Isolation And Identification Endophytic Fungi And Molecular Systematics Of Flos Lonicerae

Objective:The study on isolation and identification endophytic fungi and molecular systematics of Flos Lonicerae can provide the scientific basis for the classification and identification of endophytic fungi of Flos Lonicerae;and it supplies a theoretical basis for in-depth study and medicinal value.Methods:First, The endophytic fungi of Flos Lonicerae were isolated and purified using the tissue block method. Second, With the morphological method, we identified the endophytic fungi. The ITS sequences were extracted according to the method of molecular biology., Through the soft of blast in NCBI, analysis the sequence of the four ITS sequences. Third, Four isolated strains were analyzed and their ITS sequence were determined with the method of molecular biology. Similarty analysis was done by Blast program in GenBank. Four, With the soft of clustalx, we built the phylogenetic tree.Results: Frist, In this study, we got four pure strains. They were numbered JYH1,JYH2,JYH3 and JYH4.Second, According to the soft of Blast program in GenBank ,we matched their ITS sequences and we found the homology between JYH1 and Trichoderma aureoviride reached 99%, homology between JYH2 and Penicillium marneffei reached 99%, homology between JYH3 and Phylloporia ribis reached 97% and homology between JYH4 and Acremonium sp. reached 99%, So we can conclude that JYH1 was belongd to Trichoderma aureoviride, JYH2 was belonged to Penicillium marneffei, JYH3 was belonged to Phylloporia ribis and JYH4 was belonged to Acremonium sp..Third, Phylogeny of endophytic fungi of Flos Lonicerae and relative gerena were studied based on the sequence and other fourteen relative sequences from GenBank database, we built the phylogenetic tree. Conclusions: First, The method of molecular biology to the identification of endophytic fungi was ease, accuracy and feasible. Second, The study showed that Phylloporia ribis belong to Phylloporia and it is very near with Phellinus. Third, The four endophytic fungi of we isolated and identified have great medicinal value and base on the in-depth study.