Preliminary Study Of Fungal Diversity In Intestines Of Subadult Giant Panda

Our animals are rare and endangered giant panda, which is to maintain normal intestinal flora balance system panda gut microflora group of microorganisms, they are closely related to aspects of nutrition, metabolism, immunity and development and other hosts. Caused by intestinal flora imbalance caused by its series of intestinal microflora disease is the leading cause of death pandas. Most bacteria in the intestinal microflora, some fungi. Currently, research on panda gut microflora found in most aspects of bacterial diversity, and no use of molecular biology methods for its diversity of gut fungi. [Objective] The fungi diversity in the guts of five sub-adult giant pandaswere analyzed. [Method] Using the method of restriction fragment length polymorphism (RFLP) to analyze the fungal internal transcribed spacer sequences (ITS) of the total DNA. ITS regions were amplified with fungal universal primerstoconstruct ITS clone library. The fingerprintswere analyzed by restriction fragment length polymorphism (RFLP) of using the Hhal, HaeⅢ enzymes, respectively. The products were analyzed by sequencing and phylogenetic tree. [Results] Studies have shown that:(1) conversion of the flat from 120 randomly picked clones were analyzed, CT, YY, YX, ZM, XN respectively 105,113,107,101,106 positive clones, the positive rate was 87.5%,94.2%,89.2%,84.2%,88.3%. Was digested with restriction enzyme Hha I and Hae III for positive clones by PCR amplification products, according to the size and number of fragments stripe pattern type, the results indicate that CT, YY, YX, ZM, XN ITS intestines were separated 27,27,18,31,27 a library OTUs.Its diversity analysis, library constructed coverage above 80%, indicating that the constructed library covers a sub-adult giant panda gut most fungal species; species richness (E) were about 0.25, Shannon index (H’) were about 2.57, Simpson index (D) were about 0.11, the diversity of information tables Minya adult panda gut fungi rich; Pielou evenness index (Jsw) were about 0.79, table Minya adult panda intestinal fungal communities better distribution uniformity; according Chaol species diversity index (SChaol) predicts that subadult pandas are about 23 to 67 gut fungi.(2) the CT, YY, YX, ZM, XN OUT library belong to the same randomly selected clones were sequenced strains representing the removal of vector sequences, the resulting sequence in GenBank BLAST homology search, select the highest similarity sequence.The study showed that the gut fungi offivesub-adult giant pandas were mainly composed of Ascomycota (average of 46.24%), Basidiomycota (average of 15.79%), unclassified (average of 29.14%), unculturedfungus (average of 8.83%)Ascomycotawas mainly composed of Saccharomycetes (average of 63.74%) and Dothideomycetes (average of 35.91%); Basidiomycota was mainly composed of Tremellomycetes (average of 65.80%) and Microbotryomycetes (average of 33.15%).The four classes were mainly composed of Candida, Debaryomyces; Pleosporales, Myriangium; Cystofilobasidium, Trichosporon; Leucosporidium, Leucosporidiella. Each sample was contained with different proportion. [Conclusion] A certain proportion offungal florawere existed in the intestines of sub-adult giant pandas, and its diversity could be better analyzedby ITS-RFLP technique.