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首页> 外文期刊>Applied Microbiology >Surveying the Microbiome of Ants: Comparing 454 Pyrosequencing with Traditional Methods To Uncover Bacterial Diversity
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Surveying the Microbiome of Ants: Comparing 454 Pyrosequencing with Traditional Methods To Uncover Bacterial Diversity

机译：调查蚂蚁的微生物组：比较454焦磷酸测序与传统方法以发现细菌多样性

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We are only beginning to understand the depth and breadth of microbial associations across the eukaryotic tree of life. Reliably assessing bacterial diversity is a key challenge, and next-generation sequencing approaches are facilitating this endeavor. In this study, we used 16S rRNA amplicon pyrosequencing to survey microbial diversity in ants. We compared 454 libraries with Sanger-sequenced clone libraries as well as cultivation of live bacteria. Pyrosequencing yielded 95,656 bacterial 16S rRNA reads from 19 samples derived from four colonies of one ant species. The most dominant bacterial orders in the microbiome of the turtle ant Cephalotes varians were Rhizobiales , Burkholderiales , Opitutales , Xanthomonadales , and Campylobacterales , as revealed through both 454 sequencing and cloning. Even after stringent quality filtering, pyrosequencing recovered 445 microbe operational taxonomic units (OTUs) not detected with traditional techniques. In comparing bacterial communities associated with specific tissues, we found that gut tissues had significantly higher diversity than nongut tissues, and many of the OTUs identified from these groups clustered within ant-specific lineages, indicating a deep coevolutionary history of Cephalotes ants and their associated microbes. These lineages likely function as nutritional symbionts. One of four ant colonies investigated was infected with a Spiroplasma sp. (order Entomoplasmatales ), a potential ant pathogen. Our work shows that the microbiome associated with Cephalotes varians is dominated by a few dozen bacterial lineages and that 454 sequencing is a cost-efficient tool to screen ant symbiont diversity.
机译：我们才刚刚开始了解整个真核生物树中微生物关联的深度和广度。可靠地评估细菌多样性是一个关键的挑战，下一代测序方法正在促进这一努力。在这项研究中，我们使用16S rRNA扩增子焦磷酸测序来调查蚂蚁中的微生物多样性。我们将454个文库与Sanger测序的克隆文库以及活细菌培养进行了比较。焦磷酸测序从19种样品中获得95,656个细菌16S rRNA读数，这些样品来自一种蚂蚁物种的四个菌落。通过454测序和克隆揭示，乌龟蚂蚁Cephalotes varians的微生物组中最主要的细菌顺序是根瘤菌，Burkholderiales，Opitutales，Xanthomonadales和弯曲杆菌。即使经过严格的质量过滤，焦磷酸测序仍可恢复445种微生物操作分类单位（OTU），而传统技术则无法检测到这些分类单位。在比较与特定组织相关的细菌群落时，我们发现肠道组织比非肠道组织具有更高的多样性，并且从这些组中鉴定出的许多OTU都聚集在蚂蚁特异性谱系中，这表明头孢蚂蚁及其相关微生物具有深厚的共同进化史。 。这些谱系可能起营养共生作用。被调查的四个蚂蚁殖民地之一感染了螺旋藻。 （订购Entomoplasmatales），一种潜在的蚂蚁病原体。我们的工作表明，与头孢霉菌相关的微生物组主要由数十种细菌谱系控制，而454测序是筛选蚂蚁共生体多样性的一种经济高效的工具。

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