Agamennone Valeria, Jakupović Dennis, Weedon James T, Suring Wouter J, van Straalen Nico M, Roelofs Dick, Röling Wilfred F M
Department of Ecological Science, VU University Amsterdam, De Boelelaan 1085-1087, 1081 HV Amsterdam, the Netherlands
Department of Ecological Science, VU University Amsterdam, De Boelelaan 1085-1087, 1081 HV Amsterdam, the Netherlands.
FEMS Microbiol Ecol. 2015 Nov;91(11). doi: 10.1093/femsec/fiv128. Epub 2015 Oct 22.
The springtail Folsomia candida is an important model organism for soil ecology, ecotoxicology and ecogenomics. The decomposer activities of soil invertebrates like Folsomia depend on their relationship with microbial communities including gut symbionts. In this paper, we apply high-throughput sequencing to provide a detailed characterization of the bacterial community associated with parthenogenetic F. candida. First, we evaluated a method to suppress the amplification of DNA from the endosymbiont Wolbachia, to prevent it from interfering with the identification of less abundant operational taxonomic units (OTUs). The suppression treatment applied was effective against Wolbachia and did not interfere with the detection of the most abundant OTUs (59 OTUs, contributing over 87% of the reads). However, this method did affect the inferred community composition. Significant differences were subsequently observed in the composition of bacterial communities associated with two different strains of F. candida. A total of 832 OTUs were found, of which 45% were only present in one strain and 17% only in the other. Among the 20 most abundant OTUs, 16 were shared between strains. Denaturing gradient gel electrophoresis and clone libraries, although unable to capture the full diversity of the bacterial community, provided results that supported the NGS data.
弹尾虫秀丽隐杆线虫是土壤生态学、生态毒理学和生态基因组学的重要模式生物。像秀丽隐杆线虫这样的土壤无脊椎动物的分解活动取决于它们与包括肠道共生体在内的微生物群落的关系。在本文中,我们应用高通量测序来详细表征与孤雌生殖的秀丽隐杆线虫相关的细菌群落。首先,我们评估了一种抑制内共生菌沃尔巴克氏体DNA扩增的方法,以防止其干扰丰度较低的操作分类单元(OTU)的鉴定。所应用的抑制处理对沃尔巴克氏体有效,且不干扰最丰富的OTU(59个OTU,占读数的87%以上)的检测。然而,这种方法确实影响了推断的群落组成。随后观察到与两种不同菌株的秀丽隐杆线虫相关的细菌群落组成存在显著差异。总共发现了832个OTU,其中45%仅存在于一种菌株中,17%仅存在于另一种菌株中。在最丰富的20个OTU中,有16个在菌株之间共享。变性梯度凝胶电泳和克隆文库虽然无法捕捉细菌群落的全部多样性,但提供了支持NGS数据的结果。