Reconstructed genomes from North Pond, western flank of the Mid-Atlantic Ridge, from 2012-2014

<p>Crustal fluids were collected from the single horizon at U1382A and from the shallow, middle and deep horizons in U1383C (Edwards et al., 2012) using a mobile pumping system designed for microbial sampling from CORK fluid delivery lines as described in Meyer et al. (2016) and Cowen et al. (2012). Deployed with the ROV system, mobile pumping system connectors are attached to the CORK wellhead via an umbilical to the hydrological zone of interest within the aquifer.&nbsp; In 2012, 12 l of each fluid sample were filtered on to a 0.22 μm Sterivex-GP filter (Merck Millipore, Billerica, MA, USA) as described in Meyer et al. (2016). In 2014, 12 l of each sample was filtered in situ and immediately fixed with RNALater (Thermo Fisher Scientific, Waltham, MA, USA), as described previously (Akerman et al., 2013). After sampling in 2012, a battery-powered GeoMICROBE sled was left at each CORK for time series autonomous sampling of the fluid delivery lines (Cowen et al., 2012). For each filter sample, ~10 l of fluid were filtered in situ and immediately fixed with RNALater.&nbsp; Sleds were deployed in April 2012 and recovered in April 2014 with samples collected. Upon sled recovery, filters were transferred to fresh RNALater and stored at −80 °C, while all bag samples were stored at 4 °C (Cowen et al., 2012). Deep bottom water was sampled in 2012 and 2014 via a CTD at 100 m above the seafloor and filtered in the same manner as the crustal fluids onto Sterivex filters. Total microbial biomass in fluids was enumerated with DAPI (4′,6′-diamidino-2-phenylindole; Sigma-Aldrich, St Louis, MO, USA) and epifluorescent microscopy (Porter and Feig, 1980).&nbsp;</p>

<p>地壳流体采集自U1382A钻孔的单一地层，以及U1383C钻孔的浅、中、深三层地层（Edwards等，2012），所使用的移动泵送系统专为通过CORK流体输送管线进行微生物采样设计，其细节描述见Meyer等（2016）和Cowen等（2012）。该移动泵送系统与遥控潜水器（ROV）系统一同部署，其连接器通过脐带缆连接至CORK井口，延伸至含水层内目标水文区。2012年，每份流体样本取12升，经0.22微米Sterivex-GP滤器（美国马萨诸塞州比勒利卡市默克密理博公司）过滤，操作方法见Meyer等（2016）。2014年，每份样本取12升进行原位过滤，并立即用RNALater（美国马萨诸塞州沃尔瑟姆市赛默飞世尔科技公司）固定，方法如先前所述（Akerman等，2013）。2012年采样后，在每个CORK处留存一台电池供电的GeoMICROBE滑橇，用于对流体输送管线进行时间序列自主采样（Cowen等，2012）。对于每个滤器样本，约10升流体经原位过滤后立即用RNALater固定。滑橇于2012年4月部署，2014年4月回收并取回样本。滑橇回收后，滤器转移至新的RNALater中并于-80℃保存，而所有袋装样本则于4℃保存（Cowen等，2012）。深层底层水于2012年和2014年通过CTD（温盐深仪）在海底以上100米处采集，并采用与地壳流体相同的方法过滤至Sterivex滤器。流体中的总微生物生物量通过DAPI（4'，6'-二脒基-2-苯基吲哚；美国密苏里州圣路易斯市西格玛奥德里奇公司）染色结合落射荧光显微镜计数（Porter和Feig，1980）。</p>