[Analysis of fungal diversity in intestines of Hepialus gonggaensis larvae].
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.


CONCLUSIONS
We could get more information by combined traditional culturing and molecular biology methods.
Yu
Hewei
H
Key Lab of Genetic Function and Regulation, Bioengineering College, Chongqing University, Chongqing Engineering and Technology Center of Fungal Insecticide, Chongqing 400030, China. yuhewei54321@163.com
Wang
Zhongkang
Z
Liu
Li
L
Xia
Yuxian
Y
Yin
Youping
Y
Yuan
Qing
Q
Cao
Yueqing
Y
Peng
Guoxiong
G
D000818:Animals; D044822:Biodiversity; D018929:Cell Culture Techniques; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005658:Fungi / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D015723:Gene Library; D007422:Intestines / Q000382:microbiology*; D007814:Larva / Q000033:anatomy & histology / Q000382:microbiology; D007915:Lepidoptera / Q000033:anatomy & histology* / Q000382:microbiology*; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer