| Fungal flora refers to all the fungi that complete their life cycle or half life cycle in the soil. Soil fungi which mainly include asexual fungal is the third kind of microorganisms in microflora. Fungi’s requirement about pH is lower than actinomycetes and bacteria which makes fungi the major decomposers in acidic soil especially in forest soil.Penicillium as the most common type of saprophytes, many species of it are widely distributed in nature, almost exist everywhere, in soil, water, air, and so on. They also appear in human, animal, plant surface or in vivo, or even on other fungi. When the immunity system of humans, animals or plants is broken, some species will become opportunistic pathogens which may be harmful to health or even life. So, Penicillium is a class of fungi closed to our daily life and industrial production.Some species in Aspergillus are important and always used in the fermentation industry and food processing industry. The species in genus Aspergillus are widely distributed in the grains, air, soil, as well as a variety of organic materials. Some species of genus Aspergillus produce harmful mycotoxins, and some species can cause mold rot of fruits, vegetables and grain. Therefore, genus Aspergillus is also related to people’s life.In order to analyze the situation of soil fungi in highland areas, soil samples were collected from different habitat in the region of Kekexili, Qinghai Province and Aba, Sichuan Province. The sepatation of collected soil samples were conducted under the conditions of dilution plate method and Dicholoran Rose Bengal Chloranphenicol Media (DRBC) as a selective medium. CTAB method was used to extract the total genome DNA of strains isolated, and amplified ribosomal DNA ITS1-5.8S-ITS2 sequence, β-tubulin (BenA) partial gene sequences and calmodulin (CaM) partial gene sequence of these strains. The results obtained after comparison in Genbank were as follows:The soil samples collected from Kekexili, Qinghai were 122, and 143 strains were isolated and initially 117 strains identified after molecular identification, which are belonging to 12 genera, Penicillium:55 strains classified as 11 species, including three newly recorded species, six strains to be determined; Aspergillus:30 strains classified as nine species, including a newly recorded species, two strains to be determined; Talaromyces:one strain of one species, including a new record species; Cladosporium: two strains belong to two species, two strains to be determined; Fusarium:four strains of two species; Alternaria:one strain one species; Trichoderma:six strains classified as four species; Truncatella:three strains to be determined; Paraphoma:one strain to be determined; Embellisia:two strains belong to one species; Phoma:one strain of one species; Trichocladium:one strain to be determined.The soil samples collected from Aba, Sichuan were 64 and 188 strains were isolated, initially 162 strains identified after molecular identification, belonging to 15 genera, Penicillium:97 strains of 15 species, including six newly recorded species,27 strains to be determined; Aspergillus:six strains of four species, including a newly recorded species, two strains to be determined; Talaromyces:one strain of one species, including a new record species; Cladosporium:three strains of two species, four strains to be determined; Fusarium:one strain of one species; Alternaria:one strain of one species, one strain to be determined; Trichoderma:two strains of one species, a strain to be determined; Truncatella:six strains to be determined; Pestalotiopsis:one strain to be determined; Robillarda:three strains to be determined; Scytalidium:two strains to be determined; Tolypocladium:one strain to be determined; Wardomyces:one strain to be determined; Umbelopsis:one strain to be determined; Myrothecium:one strain to be determined.Due to the primers used to amplify calmodulin and β-tubulin gene sequences in the test were specific for Penicillium and Aspergillus, the primers may be not suitable for all the soil fungi, and the sequence analysis of ribosomal DNA ITS1-5.8S-ITS2 could not identify the strain by itself, so the strains which need to be determined require more analysis. The unidentificaed strains in this study need to be analyzed further by morphological and molecular systematics.A new record Aspergillus species, namely Aspergillus germanicus, was found in the experiments. Polyphasic studies using morphological characters and clamodulin gene, β-tubulin gene and rDNA ITS1-5.8S-ITS2 sequences. Based on the comparisions of morphological and molecular characters of the two Aspergillus isolates AS3.15303 and AS3.15304 from soil in Shandong Province, China with the ex-type of A. germanicus (CBS 123887), the two Chinese isolates were identified as A. germanicus. We confirmed that A. germanicus was a new Chinese record of Aspergillus section Usti. |
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