利用培养和免培养方法研究了一个云南红壤的真菌多样性。扩增的18S rRNA基因片段经过RsaⅠ、HinfⅠ、HaeⅢ三种限制性内切酶消化后,培养方法共获得16种限制性酶切长度多态类型(Restriction Fragment Length Polymorphism,RFLP),而免培养方法仅获得了12种RFLP类型。每个代表类型经DNA测序及系统发育分析表明,免培养方法得到的真菌全部属于子囊菌门。优势物种与Aspergillus niger类似,占免培养真菌的40.9%。其次是Penicillium属和Paecilomyces属真菌,分别占免培养真菌的18%和17%。此外,有13个克隆仅和环境克隆具有较近的亲缘关系。培养方法获取的真菌包括Ascomycota门(11.5%)、Zygomycota门(86.5%)以及Basidiomycota门(1.9%)真菌,优势克隆来自Mortierella属,占全部培养克隆的55.8%,有21.2%的序列与Absidia glauca相似。两种方法观察到的真菌物种完全不同。
The diversity of fungi in Yunnan red soil was studied through cultivation and free of cultivation methods. After the amplified 18S rRNA gene fragments were digested with RsaⅠ, HinfⅠ, and HaeⅢ restriction enzymes, the cultivation method obtained a total of 16 RFLP (Restriction Fragment Length Polymorphism), the method of free of cultivation method obtained only 12 RFLP. Analyses with DNA sequencing and phylogenesis indicated that fungi obtained with free of cuhivation were all belong to Ascomycota. And dominant species was similar to Aspergillus niger, account for 40.9% of the fungi of free of cultivation. Followed with fungi of the genus of Penicillium and the genus of Paecilomyces, respectively account for 18% and 17% of fungi of free of cultivation. Moreover, there were 13 clones fairly osculated with the environmental clones. Fungi obtained with cultivation method included Ascomycota (11.5%), Zygomycota (86.5%) , and Basidiomycota ( 1.9% ) , and the dominant clones came from the genus of Mortierella, account for 55.8% of all cultivation clones, and there were 21.2% of sequences were similar to Absidia glauca. Fungal species observed with both methods were totally different.