OBJECTIVE: Studied fungi diversity in the guts of larval Hepialus gonggaensis using culture-independent and traditional culturing methods. METHODS: For the culture-independent method, the total DNA of fungus was extracted from the intestinal contents and internal transcribed spacer (ITS) regions were amplified with fungal universal primers. A near-full length ITS gene library was constructed. Subsequently, the fingerprints of the microorganisms were analyzed by isolated plasmid and digestion with Msp I, HaeIII and Taq I enzymes, respectively. Restriction fragment length polymorphism (RFLP) analysis based on the fungal ITS sequences indicated that the library established includes 23 operational taxonomic units (OTUS) and a phylogenetic tree depicted the linkage of the isolated fungi. RESULTS: Abundant fungi were in the intestines of H. gonggaensis larvae, but their abundance was very different. The dominant fungi belonged to Mortierellaone and Trichosporon and accounted for 46.34 % and 39.02 % of the total ITS clones, respectively. Only three genera of fungi were identified from eight isolated fungal populations by traditional culturing methods. CONCLUSION: We could get more information by combined traditional culturing and molecular biology methods.
OBJECTIVE: Studied fungi diversity in the guts of larval Hepialus gonggaensis using culture-independent and traditional culturing methods. METHODS: For the culture-independent method, the total DNA of fungus was extracted from the intestinal contents and internal transcribed spacer (ITS) regions were amplified with fungal universal primers. A near-full length ITS gene library was constructed. Subsequently, the fingerprints of the microorganisms were analyzed by isolated plasmid and digestion with Msp I, HaeIII and Taq I enzymes, respectively. Restriction fragment length polymorphism (RFLP) analysis based on the fungal ITS sequences indicated that the library established includes 23 operational taxonomic units (OTUS) and a phylogenetic tree depicted the linkage of the isolated fungi. RESULTS: Abundant fungi were in the intestines of H. gonggaensis larvae, but their abundance was very different. The dominant fungi belonged to Mortierellaone and Trichosporon and accounted for 46.34 % and 39.02 % of the total ITS clones, respectively. Only three genera of fungi were identified from eight isolated fungal populations by traditional culturing methods. CONCLUSION: We could get more information by combined traditional culturing and molecular biology methods.