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METAGENOMIC ANALYSIS OF SOIL MICROBIAL COMMUNITIES

机译:土壤微生物群落的计量经济学分析

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Ramonda serbica and Ramonda nathaliae, rare resurrection plants growing in the Balkan Peninsula, produce a high amount of phenolic compounds as a response to stress. The composition and size of bacterial communities in two rhizosphere soil samples of these plants were analyzed using a metagenomic approach. Fluorescent in situ hybridization (FISH) experiments together with DAPI staining showed that the metabolically active bacteria represent only a small fraction, approximately 5%, of total soil bacteria. Using universal bacteria – specific primers 16S rDNA genes were amplified directly from metagenomic DNAs and two libraries were constructed. The Restriction Fragment Length Polymorphism (RLFP) method was used in library screening. Amongst 192 clones, 35 unique operational taxonomic units (OTUs) were determined from the rhizosphere of R. nathaliae, and 13 OTUs out of 80 clones in total from the library of R. serbica. Representative clones from each OTU were sequenced. The majority of sequences from metagenomes showed very little similarity to any cultured bacteria. In conclusion, the bacterial communities in the studied soil samples showed quite poor diversity.
机译:巴尔干半岛上罕见的复活植物Ramonda serbica和Ramonda nathaliae会产生大量酚类化合物,以应对压力。使用宏基因组学方法分析了这些植物的两个根际土壤样品中细菌群落的组成和大小。荧光原位杂交(FISH)实验和DAPI染色表明,代谢活性细菌仅占土壤细菌总数的一小部分,约占5%。使用通用细菌–直接从宏基因组DNA中扩增特异性引物16S rDNA基因,并构建了两个文库。限制性片段长度多态性(RLFP)方法用于文库筛选。在192个克隆中,从nathaliae根际中确定了35个独特的操作分类单位(OTU),在来自R. serbica库的总共80个克隆中确定了13个OTU。对来自每个OTU的代表性克隆进行测序。来自元基因组的大多数序列显示与任何培养细菌几乎没有相似性。总之,研究土壤样品中的细菌群落显示出很差的多样性。

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