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首页> 外文期刊>Applied Microbiology >Pyrosequencing-Derived Bacterial, Archaeal, and Fungal Diversity of Spacecraft Hardware Destined for Mars
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Pyrosequencing-Derived Bacterial, Archaeal, and Fungal Diversity of Spacecraft Hardware Destined for Mars

机译：焦磷酸测序的细菌，古细菌和真菌多样性，发往火星的航天器硬件

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Spacecraft hardware and assembly cleanroom surfaces (233 m~(2) in total) were sampled, total genomic DNA was extracted, hypervariable regions of the 16S rRNA gene (bacteria and archaea) and ribosomal internal transcribed spacer (ITS) region (fungi) were subjected to 454 tag-encoded pyrosequencing PCR amplification, and 203,852 resulting high-quality sequences were analyzed. Bioinformatic analyses revealed correlations between operational taxonomic unit (OTU) abundance and certain sample characteristics, such as source (cleanroom floor, ground support equipment [GSE], or spacecraft hardware), cleaning regimen applied, and location about the facility or spacecraft. National Aeronautics and Space Administration (NASA) cleanroom floor and GSE surfaces gave rise to a larger number of diverse bacterial communities (619 OTU; 20 m~(2)) than colocated spacecraft hardware (187 OTU; 162 m~(2)). In contrast to the results of bacterial pyrosequencing, where at least some sequences were generated from each of the 31 sample sets examined, only 13 and 18 of these sample sets gave rise to archaeal and fungal sequences, respectively. As was the case for bacteria, the abundance of fungal OTU in the GSE surface samples dramatically diminished (9× less) once cleaning protocols had been applied. The presence of OTU representative of actinobacteria, deinococci, acidobacteria, firmicutes, and proteobacteria on spacecraft surfaces suggests that certain bacterial lineages persist even following rigorous quality control and cleaning practices. The majority of bacterial OTU observed as being recurrent belonged to actinobacteria and alphaproteobacteria, supporting the hypothesis that the measures of cleanliness exerted in spacecraft assembly cleanrooms (SAC) inadvertently select for the organisms which are the most fit to survive long journeys in space.
机译：采样航天器的硬件和组装洁净室表面（共233 m〜（2）），提取总基因组DNA，提取16S rRNA基因的高变区（细菌和古细菌）和核糖体内部转录间隔区（ITS）区（真菌）。进行454标签编码的焦磷酸测序PCR扩增，并对203,852个高质量序列进行了分析。生物信息学分析显示，操作分类单元（OTU）的丰度与某些样品特征之间的相关性，例如来源（洁净室地板，地面支撑设备[GSE]或航天器硬件），所应用的清洁方案以及设施或航天器的位置。美国国家航空航天局（NASA）的洁净室地板和GSE表面产生了比共置航天器硬件更大的细菌群落（619 OTU; 20 m〜（2））（187 OTU; 162 m〜（2））。与细菌焦磷酸测序的结果相反，从所检查的31个样本集中，至少每个产生了一些序列，而这些样本集中只有13和18个分别产生了古细菌和真菌序列。与细菌一样，一旦应用了清洁方案，GSE表面样品中真菌OTU的含量就会大大降低（减少了9倍）。在航天器表面上存在代表放线菌，脱球菌，酸性细菌，硬毛和蛋白细菌的OTU，这表明即使严格的质量控制和清洁措施，某些细菌谱系仍然存在。被观察到复发的大多数细菌OTU属于放线菌和α变形杆菌，支持以下假设：在航天器组装洁净室（SAC）中采用的清洁措施无意中选择了最适合在太空中长期生存的生物。

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