| Located at Zoige county, Aba Tibetan Autonomous Prefecture, Sichuan Province, Zoige wetland occupies an important position at the wetland biodiversity protection in the Qinghai Tibet Plateau. Fungal diversity, as an important part of Ruoergai wetland species diversity, has great research value. Collecting a total of 100 soil samples in Ruoergai wetland, the paper studies soil fungal diversity and ecological niche of Ruoergai wetland by the means of dilution plate and Miseq high throughput sequencing.The result taking use of dilution plate to test 100 soil samples shows that the average number of soil fungi in Zoige wetland is 237.86±49.68(1.05?3705.4)cfu.g-and the number of fungi in the 4 soil types is following as:the subalpine meadow soil (446.27+195.92 cfu.g-1)> meadow soil (304.84+206.20 cfu.g-1)> aeolian sandy soil (136.09+49.05 cfu.g-1)> peat soil (132.95+50.57 cfu.g-1). A total of 593 fungus strains were isolated.337 strains of fungi were identified, in which includes ascomycota, basidiomycota and zygomycota. There were 249 strains of Ascomycota, accounting for 73.88%, including Penicillium spp., Trichoderma spp., Aspergillus spp., Fusarium sp., Alternaria sp., Vertieillum sp., Phoma spp., Pestalotiopsis sp., Trichocladium sp., Chaetomium spp., Myrothecium sp., Cladosporium sp., Cordyceps sp., Epicoccum sp., Didymella sp., Paraphaeosphaeria spp., Oidiodendron sp., Nectria sp., Trichocladium spp, Meyerozyma spp. and Mortierella spp.. There were 85 strains of basidiomycota, accounting for25.22%, including Mucor spp., Rhizopus spp., Mortierella spp., Actinomucor spp. And Zygorhynchus sp.. There were 3 strains of zygomycota, accounting for 0.89%, including Schizophyllum sp.. The relative abundance of M.hiemalis was 50.11% in aeolian sandy soil, which made it a dominant group. In Zoige wetland soil fungi, the frequency of separation among the top 5 are Mucor hiemalis, Penicillium subrubescens, Penicillium sp., Cladosporium sphaerospermum and Trichoderma harzianum. Penicillium dipodomyicola and Penicillium subrubescens are dominant groups in meadow soil, with the relative abundance of 34.43% and 11.56% respectively. P. subrubescens and Penicillium sp are dominant groups in subalpine meadow soil, with the relative abundance of 20.43% and 18.14% respectively. Cladosporium sphaerospermum and Trichoderma harzianum are dominant groups in peat soil, with the relative abundance of 20.99% and 20.22% respectively.The average diversity index of Zoige wetlands fungi is 1.03±0.06 (0-2.75) average richness index 5.82±0.40 (1-24), and the average of evenness index 0.63±0.02 (0.06-1), then the diversity index of the 4 kinds of soil is as following: aeolian sandy soil (1.24+0.24)> subalpine meadow soil (1.02+0.12)> meadow soil (0.89+0.12)> peat soil (0.65+0.18). The richness index of the 4 kinds of soil types is shown as aeolian sandy soil (9+1.69)> subalpine meadow soil (5.31+0.53)> meadow soil (4.5+0.47)> peat soil (3.2+0.61). The evenness index of the 4 kinds of soil types is described as peat soil (0.69+0.11)> sub alpine meadow soil (0.61+ 0.05)> meadow soil (0.59+0.05)> aeolian sandy soil (0.58+0.09). For the soil type determiners, the ecological niche of Penicillium subrubescens, Mucor hiemalis and Trichoderma harzianum are wider. For the soil depth determiner, the ecological niche of Penicillium skrjabinii, Penicillium subrubescens, Penicillium, Penicillium dipodomyicola, Fusarium sp., Cladosporium sphaerospermum, Actinomucor elegans, Mucor hiemalis, Mortierella alpine and Trichoderma saturnisporum are wider.High throughput sequencing was carried out on 11 samples of the 4 soil types, the average Reads number per sample was 4127655337. The total number of detected fungi is 701 species, belonging to 385 genera, including Zygomycota, Ascomycota, Chytridiomycota, Basidiomycota, Glomeromycota, and Blastocladiomycota. Blastocladiomycota only survive in subalpine meadow soil. At the species level classification, species detected in meadow soil is 42, in aeolian sandy soil 59, and in the peat soil 64. Result of PCoA (Principal Co-ordinates Analysis) shows that samples of different soil types have a common intersection. If the average diversity index of soil fungi in Zoige wetland is 7.76+0.31 (6.11-9.3), and the average OTU richness 3569+341.81 (1536-5324), then diversity index of 4 soil types showes peat soil (8.80+0.50)> sub alpine meadow soil (8.34+0.23)> meadow soil (7.19+ 0.71)> aeolian sandy soil (7.05+0.48). The richness of OTU in the 4 soil types is expressed as peat soil (4063.50+9.50)> meadow soil (3789.67+516.18)> sub alpine meadow soil (3530+1017.25)> aeolian sandy soil (3057.67). For the soil type determiner, ecological niche of Penicillum, Fusarium, Aspergillus, Cladosporium, Phoma, Mortierella, Chaetomium, Coryespora, Preussia, Cryptococcus and Glomus is wider, which shows that the preference for resources is small, and the adaptability to soil types is wider.Comparing with the results of the dilution plate and high throughput sequencing, we find that high throughput sequencing provides greater information than the traditional soil fungal isolation method, and the fungal diversity detected by it is much higher than the dilution plate.13 genera of top 20 for the relative abundance detected the high throughput sequencing are not separated by the traditional method of separation. However, common soil fungi, such as wood mold could not be measured by the high throughput sequencing. For the calculation of ecological niche, comparing with the traditional separation of cultivating method and high-throughput sequencing, we find that the ecological niche of Penicillium spp, Fusarium sp., Aspergillus spp., Cladosporium sp., Alternaria sp., and Mortierella.alpina spp. measured by high-throughput sequencing are much higher. In conclusion, high throughput sequencing is a more accurate and efficient method for the study of soil microbial ecology. |
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