Data repository of "How do male danaine butterflies locate pyrrolizidine alkaloids?: Foreleg as an olfactory alkaloid-sensing organ"

This dataset contains results from behavioral and electrophysiological experiments

A Long-Term Cultivation of an Anaerobic Methane-Oxidizing Microbial Community from Deep-Sea Methane-Seep Sediment Using a Continuous-Flow Bioreactor

<div><p>Anaerobic oxidation of methane (AOM) in marine sediments is an important global methane sink, but the physiological characteristics of AOM-associated microorganisms remain poorly understood. Here we report the cultivation of an AOM microbial community from deep-sea methane-seep sediment using a continuous-flow bioreactor with polyurethane sponges, called the down-flow hanging sponge (DHS) bioreactor. We anaerobically incubated deep-sea methane-seep sediment collected from the Nankai Trough, Japan, for 2,013 days in the bioreactor at 10°C. Following incubation, an active AOM activity was confirmed by a tracer experiment using ¹³C-labeled methane. Phylogenetic analyses demonstrated that phylogenetically diverse <i>Archaea</i> and <i>Bacteria</i> grew in the bioreactor. After 2,013 days of incubation, the predominant archaeal components were <u>an</u>aerobic <u>me</u>thanotroph (ANME)-2a, Deep-Sea Archaeal Group, and Marine Benthic Group-D, and <i>Gammaproteobacteria</i> was the dominant bacterial lineage. Fluorescence <i>in situ</i> hybridization analysis showed that ANME-1 and -2a, and most ANME-2c cells occurred without close physical interaction with potential bacterial partners. Our data demonstrate that the DHS bioreactor system is a useful system for cultivating fastidious methane-seep-associated sedimentary microorganisms.</p></div

Statistical analysis of clone libraries.

a<p>A phylotype was defined as ≥97% sequence identity.</p>b<p>Numbers in parentheses indicate the 95% confidence interval.</p

(A and G–I) FISH and (B–F) CARD-FISH images of microbial cells cultivated in the DHS bioreactor.

<p>Photomicrographs of DAPI-stained cells (left) and epifluorescence (right) showing identical fields. (A) Chain-forming ANME-1-350-stained cells in the 2,013-day sample (Alexa Fluor 488, green). (B) An ANME-2a-647-stained coccoid-shaped cell in the 2,013-day sample (Fluorescein, green). (C and D) Color overlay of ANME-2c-760- (Fluorescein, green) and EUB338-stained cells (Alexa Fluor 594, red) in the 903-day sample. (E) A MBGB-380-stained coccoid-shaped cell in the 2,013-day sample (Fluorescein, green). (F) A MBGD-318-stained rod-shaped cell in the 2,013-day sample (Fluorescein, green). (G) A MCOCID442-stained coccoid-shaped cell in the 2,013-day sample. (H) Mγ669-stained coccoid-shaped cells in the 2,013-day sample (Alexa Fluor 488, green). (I) UncGAM731-stained irregular rod-shaped cells in the 2,013-day sample (Alexa Fluor 488, green). Bars represent 10 µm.</p

Comparison of AOM enrichments in different types of continuous-flow bioreactors.

<p>NR: not reported.</p>a<p>Data from long-term incubations with short columns described in Wegener and Boetius <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105356#pone.0105356-Wegener1" target="_blank">[22]</a>.</p>b<p>Data from high-flow experiments with NON-SEEP sediments described in Girguis <i>et al</i>. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105356#pone.0105356-Girguis2" target="_blank">[25]</a>.</p>c<p>Data from new high flow core experiments described in Steeb <i>et al</i>. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105356#pone.0105356-Steeb1" target="_blank">[23]</a>.</p>d<p>Dominant ANME type is shown in bold.</p