利用变性梯度凝胶电泳分析正红菇菌根围土壤真菌群落多样性

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以正红菇(Russula griseocarnosa)菌根围土壤为研究对象,通过提取土壤基因组DNA,以通用引物扩增真菌18S rRNA基因V1+V2区,将PCR产物进行变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis),获得土壤微生物群落的DNA特征指纹图谱,并对图谱中的优势条带回收测序,通过Blast进行同源性比对并构建系统发育树,进而分析正红菇菌根围真菌群落组成及多样性。同源性比对结果表明,在回收测序的19条DGGE条带中,4条为非真菌真核生物序列,系统发育分析显示全部序列可以分为4类菌群,GroupⅠ主要为担子菌门(Basidiomycota)真菌,GroupⅡ主要为子囊菌门(Ascomycota)真菌,GroupⅢ为未知真菌,GroupⅣ主要为节肢动物门生物(Arthropoda)。 The rhizosphere soil of Russula griseocarnosa was used as the research object. The genomic DNA of soil was extracted, and the V1 + V2 region of 18S rRNA gene of fungus was amplified by universal primers. The PCR products were subjected to denaturing gradient gel electrophoresis ) To obtain the DNA fingerprint of the soil microbial community. The dominant bands in the map were recovered and sequenced. The homology comparison was made by Blast and the phylogenetic tree was constructed. Then the composition and diversity Sex. Homology comparison showed that among the 19 DGGE bands recovered and sequenced, 4 were non-eukaryotic eukaryotic sequences. Phylogenetic analysis showed that all the sequences could be divided into 4 groups. GroupⅠ mainly consisted of basidiomycota Basidiomycota fungi. Group II is mainly composed of Ascomycota fungi, Group III is an unknown fungus and Group IV is mainly Arthropoda.
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