垃圾填埋场中厌氧真菌18SrDNA的PCR扩增及鉴定

采用机械破壁法直接从来自7个不同地区的垃圾填埋场滤液样本中提取真菌DNA，应用真菌通用引物NS1和NS8扩增18SrDNA（约1800bp)，多聚酶链式反应（PCR）产物的琼脂糖凝胶电泳结果表明所有的样本均得到了扩增；以PCR产物作为模板，采用厌氧真菌Chytridiomycetes科的专用引物Chyt-719和Chyt-1553进行二次PCR扩增（约857bp)，该阳性扩增产物克隆和测序结果首次表明在食草动物瘤胃中存在的厌氧真菌Chytridiomycetes也存在于垃圾填埋场中，且为Neocallimastix属。

PCR Amplification of Anaerobic Fungal 18S rDNA from Landfill Sites

Nucleic acid based techniques are widely used to characterize microbial communities in environmental samples.In this study,seven landfill leachates were tested for the presence of 18S rRNA genes from anaerobic fungi from the family Chytridiomycetes.A nested Polymerase Chain Reaction (PCR) strategy was employed to amplify the 18S rRNA gene region of Chytridiomycetous fungi from landfill sites samples.All of the PCR products from landfill leachates amplified with the first primer pair NS1/NS8 and the second primer pair Chyt\|719/Chyt\|1553 were subjected to agarose gel electrophoresis to show seven samples containing fungi from the family Chytridiomycetes.One of the positive results was then selected for cloning and sequencing of the PCR product to confirm the identity of the amplified DNA.The molecular data presented here reveals for the first time that Chytridiomycetous fungi associated with the anaerobic rumen environment are also present in landfill sites.