Abstract
Coarse woody debris is an important biomass pool in forest ecosystems that numerous groups of insects have evolved to take advantage of. These insects are ecologically important and represent useful natural analogs for biomass to biofuel conversion. Using a range of molecular approaches combined with microelectrode measurements of oxygen, we have characterized the gut microbiome and physiology of Odontotaenius disjunctus, a wood-feeding beetle native to the eastern United States. We hypothesized that morphological and physiological differences among gut regions would correspond to distinct microbial populations and activities. In fact, significantly different communities were found in the foregut (FG), midgut (MG)/posterior hindgut (PHG) and anterior hindgut (AHG), with Actinobacteria and Rhizobiales being more abundant toward the FG and PHG. Conversely, fermentative bacteria such as Bacteroidetes and Clostridia were more abundant in the AHG, and also the sole region where methanogenic Archaea were detected. Although each gut region possessed an anaerobic core, micron-scale profiling identified radial gradients in oxygen concentration in all regions. Nitrogen fixation was confirmed by 15N2 incorporation, and nitrogenase gene (nifH) expression was greatest in the AHG. Phylogenetic analysis of nifH identified the most abundant transcript as related to Ni–Fe nitrogenase of a Bacteroidetes species, Paludibacter propionicigenes. Overall, we demonstrate not only a compartmentalized microbiome in this beetle digestive tract but also sharp oxygen gradients that may permit aerobic and anaerobic metabolism to occur within the same regions in close proximity. We provide evidence for the microbial fixation of N2 that is important for this beetle to subsist on woody biomass.
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Acknowledgements
This work was supported by the Department of Energy, Office of Biological and Environmental Research, Genomic Sciences program through the Lawrence Livermore National Laboratory Biofuels Scientific Focus Area (SFA) award SCW1039. Part of this work (ELB, JAC-N, UK, GLA) was performed at Lawrence Berkeley National Laboratory under the Department of Energy contract number DE-AC02-05CH11231. Contributions of JPR are under the auspices of the US DOE at LLNL (DE-AC52-07NA27344). Additional support was provided from the NSF-GRFP program to NHN, the LSU Boyd Professor Research fund to MB, and JAC-N was also supported in part by a grant from ‘Consejo Nacional de Ciencia y Tecnología’ (CONACyT, Mexico). We thank Hector Urbina for collection of beetles; George Stanley for help in conducting the nitrogen uptake experiments; Doug Wendell and Gail Ackermann form the QIIME-DB for their support with the submission of the sequencing data to the EBI.
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Ceja-Navarro, J., Nguyen, N., Karaoz, U. et al. Compartmentalized microbial composition, oxygen gradients and nitrogen fixation in the gut of Odontotaenius disjunctus. ISME J 8, 6–18 (2014). https://doi.org/10.1038/ismej.2013.134
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DOI: https://doi.org/10.1038/ismej.2013.134
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