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Comparative transcriptomic insights into the mechanisms of electron transfer in Geobacter co-cultures with activated carbon and magnetite

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Abstract

Both activated carbon and magnetite have been reported to promote the syntrophic growth of Geobacter metallireducens and Geobacter sulfurreducens co-cultures, the first model to show direct interspecies electron transfer (DIET); however, differential transcriptomics of the promotion on co-cultures with these two conductive materials are unknown. Here, the comparative transcriptomic analysis of G. metallireducens and G. sulfurreducens co-cultures with granular activated carbon (GAC) and magnetite was reported. More than 2.6-fold reduced transcript abundances were determined for the uptake hydrogenase genes of G. sulfurreducens as well as other hydrogenases in those co-cultures to which conductive materials had been added. This is consistent with electron transfer in G. metallireducens-G. sulfurreducens co-cultures as evinced by direct interspecies electron transfer (DIET). Transcript abundance for the structural component of electrically conductive pili (e-pili), PilA, was 2.2-fold higher in G. metallireducens, and, in contrast, was 14.9-fold lower in G. sulfurreducens in co-cultures with GAC than in Geobacters co-cultures without GAC. However, it was 9.3-fold higher in G. sulfurreducens in co-cultures with magnetite than in Geobacters co-cultures. Mutation results showed that GAC can be substituted for the e-pili of both strains but magnetite can only compensate for that of G. sulfurreducens, indicating that the e-pili is a more important electron acceptor for the electron donor strain of G. metallireducens than for G. sulfurreducens. Transcript abundance for G. metallireducens c-type cytochrome gene GMET_RS14535, a homologue to c-type cytochrome gene omcE of G. sulfurreducens was 9.8-fold lower in co-cultures with GAC addition, while that for OmcS of G. sulfurreducens was 25.1-fold higher in co-cultures with magnetite, than in that without magnetite. Gene deletion studies showed that neither GAC nor magnetite can completely substitute the cytochrome (OmcE homologous) of G. metallireducens but compensate for the cytochrome (OmcS) of G. sulfurreducens. Moreover, some genes associated with central metabolism were up-regulated in the presence of both GAC and magnetite; however, tricarboxylic acid cycle gene transcripts in G. sulfurreducens were not highly-expressed in each of these amended co-cultures, suggesting that there was considerable redundancy in the pathways utilised by G. sulfurreducens for electron transfer to reduce fumarate with the amendment of GAC or magnetite. These results support the DIET model of G. metallireducens and G. sulfurreducens and suggest that e-pili and cytochromes of the electron donor strain are more important than that of the electron acceptor strain, indicating that comparative transcriptomics may be a promising route by which to reveal different responses of electron donor and acceptor during DIET in co-cultures.

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References

  • Aklujkar, M., Coppi, M.V., Leang, C., Kim, B.C., Chavan, M.A., Perpetua, L.A., Giloteaux, L., Liu, A., and Holmes, D.E. (2013). Proteins involved in electron transfer to Fe(III) and Mn(IV) oxides by Geobacter sulfurreducens and Geobacter uraniireducens. Microbiology 159, 515–535.

    Article  PubMed  CAS  Google Scholar 

  • Benjamini, Y., and Yekutieli, D. (2001). The control of the false discovery rate in multiple testing under dependency. Ann Statist 29, 1165–1188.

    Article  Google Scholar 

  • Burrows, L.L. (2012). Pseudomonas aeruginosa twitching motility: type IV pili in action. Annu Rev Microbiol 66, 493–520.

    Article  PubMed  CAS  Google Scholar 

  • Chen, S., Rotaru, A.E., Liu, F., Philips, J., Woodard, T.L., Nevin, K.P., and Lovley, D.R. (2014a). Carbon cloth stimulates direct interspecies electron transfer in syntrophic co-cultures. Bioresour Tech 173, 82–86.

    Article  CAS  Google Scholar 

  • Chen, S., Rotaru, A.E., Shrestha, P.M., Malvankar, N.S., Liu, F., Fan, W., Nevin, K.P., and Lovley, D.R. (2014b). Promoting interspecies electron transfer with biochar. Sci Rep 4, 5019.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Childers, S.E., Ciufo, S., and Lovley, D.R. (2002). Geobacter metallireducens accesses insoluble Fe(iii) oxide by chemotaxis. Nature 416, 767–769.

    Article  PubMed  CAS  Google Scholar 

  • Coppi, M.V., Leang, C., Sandler, S.J., and Lovley, D.R. (2001). Development of a genetic system for Geobacter sulfurreducens. Appl Environ Microbiol 67, 3180–3187.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Coppi, M.V., O’Neil, R.A., and Lovley, D.R. (2004). Identification of an uptake hydrogenase required for hydrogen-dependent reduction of Fe(III) and other electron acceptors by Geobacter sulfurreducens. J Bacteriol 186, 3022–3028.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Holmes, D.E., Dang, Y., Walker, D.J.F., and Lovley, D.R. (2016). The electrically conductive pili of Geobacter species are a recently evolved feature for extracellular electron transfer. Microbial Genomics 2, e000072.

    Article  PubMed  PubMed Central  Google Scholar 

  • Kato, S., Hashimoto, K., and Watanabe, K. (2012). Microbial interspecies electron transfer via electric currents through conductive minerals. Proc Natl Acad Sci USA 109, 10042–10046.

    Article  PubMed  Google Scholar 

  • Kato, S., Hashimoto, K., and Watanabe, K. (2013). Iron-oxide minerals affect extracellular electron-transfer paths of Geobacter spp.. Microb Environ 28, 141–148.

    Article  Google Scholar 

  • Leang, C., Qian, X., Mester, T., and Lovley, D.R. (2010). Alignment of the c-type cytochrome OmcS along Pili of Geobacter sulfurreducens. Appl Environ MicroBiol 76, 4080–4084.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Li, R., Yu, C., Li, Y., Lam, T.W., Yiu, S.M., Kristiansen, K., and Wang, J. (2009). SOAP2: an improved ultrafast tool for short read alignment. Bioinformatics 25, 1966–1967.

    Article  PubMed  CAS  Google Scholar 

  • Liu, F., Rotaru, A.E., Shrestha, P.M., Malvankar, N.S., Nevin, K.P., and Lovley, D.R. (2012). Promoting direct interspecies electron transfer with activated carbon. Energ Environ Sci 5, 8982–8989.

    Article  CAS  Google Scholar 

  • Liu, F., Rotaru, A.E., Shrestha, P.M., Malvankar, N.S., Nevin, K.P., and Lovley, D.R. (2015). Magnetite compensates for the lack of a pilin-associated c-type cytochrome in extracellular electron exchange. Environ Microbiol 17, 648–655.

    Article  PubMed  CAS  Google Scholar 

  • Mahadevan, R., Palsson, B.Ø., and Lovley, D.R. (2011). In situ to in silico and back: elucidating the physiology and ecology of Geobacter spp. using genome-scale modelling. Nat Rev Micro 9, 39–50.

    Article  CAS  Google Scholar 

  • Malvankar, N.S., and Lovley, D.R. (2014). Microbial nanowires for bioenergy applications. Curr Opin Biotech 27, 88–95.

    Article  PubMed  CAS  Google Scholar 

  • Malvankar, N.S., Tuominen, M.T., and Lovley, D.R. (2012). Lack of cytochrome involvement in long-range electron transport through conductive biofilms and nanowires of Geobacter sulfurreducens. Energ Environ Sci 5, 8651–8659.

    Article  CAS  Google Scholar 

  • Malvankar, N.S., Vargas, M., Nevin, K., Tremblay, P.L., Evans-Lutterodt, K., Nykypanchuk, D., Martz, E., Tuominen, M.T., and Lovley, D.R. (2015). Structural basis for metallic-like conductivity in microbial nanowires. MBio 6, e00084–15.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Malvankar, N.S., Vargas, M., Nevin, K.P., Franks, A.E., Leang, C., Kim, B.C., Inoue, K., Mester, T., Covalla, S.F., Johnson, J.P., Rotello, V.M., Tuominen, M.T., and Lovley, D.R. (2011). Tunable metallic-like conductivity in microbial nanowire networks. Nat Nanotech 6, 573–579.

    Article  Google Scholar 

  • McCallum, M., Tammam, S., Khan, A., Burrows, L.L., and Howell, P.L. (2017). The molecular mechanism of the type IVa pilus motors. Nat Commun 8, 15091.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Mehta, T., Coppi, M.V., Childers, S.E., and Lovley, D.R. (2005). Outer membrane c-type cytochromes required for Fe(III) and Mn(IV) oxide reduction in Geobacter sulfurreducens. Appl Environ MicroBiol 71, 8634–8641.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Mortazavi, A., Williams, B.A., McCue, K., Schaeffer, L., and Wold, B. (2008). Mapping and quantifying mammalian transcriptomes by RNASeq. Nat Meth 5, 621–628.

    Article  CAS  Google Scholar 

  • Okamoto, A., Saito, K., Inoue, K., Nealson, K.H., Hashimoto, K., and Nakamura, R. (2014). Uptake of self-secreted flavins as bound cofactors for extracellular electron transfer in Geobacter species. Energ Environ Sci 7, 1357–1361.

    Article  CAS  Google Scholar 

  • Pokkuluri, P.R., Londer, Y.Y., Yang, X., Duke, N.E.C., Erickson, J., Orshonsky, V., Johnson, G., and Schiffer, M. (2010). Structural characterization of a family of cytochromes c7 involved in Fe(III) respiration by Geobacter sulfurreducens. Biochim Biophys Acta 1797, 222–232.

    Article  PubMed  CAS  Google Scholar 

  • Reguera, G., McCarthy, K.D., Mehta, T., Nicoll, J.S., Tuominen, M.T., and Lovley, D.R. (2005). Extracellular electron transfer via microbial nanowires. Nature 435, 1098–1101.

    Article  PubMed  CAS  Google Scholar 

  • Rotaru, A.E., Shrestha, P.M., Liu, F., Ueki, T., Nevin, K., Summers, Z.M., and Lovley, D.R. (2012). Interspecies electron transfer via hydrogen and formate rather than direct electrical connections in cocultures of Pelobacter carbinolicus and Geobacter sulfurreducens. Appl Environ MicroBiol 78, 7645–7651.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Shrestha, P.M., Kube, M., Reinhardt, R., and Liesack, W. (2009). Transcriptional activity of paddy soil bacterial communities. Environ Microbiol 11, 960–970.

    Article  PubMed  CAS  Google Scholar 

  • Shrestha, P.M., Rotaru, A.E., Summers, Z.M., Shrestha, M., Liu, F., and Lovley, D.R. (2013). Transcriptomic and genetic analysis of direct interspecies electron transfer. Appl Environ Microbiol 79, 2397–2404.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Smith, J.A., Lovley, D.R., and Tremblay, P.L. (2013). Outer cell surface components essential for Fe(III) oxide reduction by Geobacter metallireducens. Appl Environ Microbiol 79, 901–907.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Summers, Z.M., Fogarty, H.E., Leang, C., Franks, A.E., Malvankar, N.S., and Lovley, D.R. (2010). Direct exchange of electrons within aggregates of an evolved syntrophic coculture of anaerobic bacteria. Science 330, 1413–1415.

    Article  PubMed  CAS  Google Scholar 

  • Tan, Y., Adhikari, R.Y., Malvankar, N.S., Ward, J.E., Woodard, T.L., Nevin, K.P., and Lovley, D.R. (2017). Expressing the Geobacter metallireducens PilA in Geobacter sulfurreducens yields pili with exceptional conductivity. MBio 8, e02203–16.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Thiele, J.H., and Zeikus, J.G. (1988). Control of interspecies electron flow during anaerobic-digestion: significance of formate transfer versus hydrogen transfer during syntrophic methanogenesis in flocs. Appl Environ Microbiol 54, 20–29.

    PubMed  PubMed Central  CAS  Google Scholar 

  • Thorpe, C.L., Morris, K., Boothman, C., and Lloyd, J.R. (2012). Alkaline Fe (III) reduction by a novel alkali-tolerant Serratia sp. isolated from surface sediments close to Sellafield nuclear facility, UK. FEMS Microbiol Lett 327, 87–92.

    Article  PubMed  CAS  Google Scholar 

  • Tremblay, P.L., Aklujkar, M., Leang, C., Nevin, K.P., and Lovley, D. (2012). A genetic system for Geobacter metallireducens: role of the flagellin and pilin in the reduction of Fe(III) oxide. Environ Microbiol Rep 4, 82–88.

    Article  PubMed  CAS  Google Scholar 

  • Tremblay, P.L., and Lovley, D.R. (2012). Role of the NiFe hydrogenase Hya in oxidative stress defense in Geobacter sulfurreducens. J Bacteriol 194, 2248–2253.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Vargas, M., Malvankar, N.S., Tremblay, P.L., Leang, C., Smith, J.A., Patel, P., Snoeyenbos-West, O., Nevin, K.P., and Lovley, D.R. (2013). Aromatic amino acids required for Pili conductivity and long-range extracellular electron transport in Geobacter sulfurreducens. MBio 4, e00210-13–e00210-13.

    Article  PubMed Central  Google Scholar 

  • Zheng, S., Zhang, H., Li, Y., Zhang, H., Wang, O., Zhang, J., and Liu, F. (2015). Co-occurrence of Methanosarcina mazei and Geobacteraceae in an iron (III)-reducing enrichment culture. Front Microbiol 6, 941.

    PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the Major Research plan (91751112) and the General Programme (41371257, 41573071) of the National Natural Science Foundation of China, Shandong Natural Science Fund for Distinguished Young Scholars (JQ201608), the Young Taishan Scholars Programme of Shandong Province (tsqn20161054) and the Key Research Project for Frontier Science of the Chinese Academy of Sciences (QYZDJ-SSW-DQC015).

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Correspondence to Fanghua Liu or Oumei Wang.

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Table S1

Statistical data of unique Illumina sequence reads of G. metallireducens and G. sulfurreducens transcripts from co-cultures of G.m_G.s, G.m_G.s_GAC, and G.m_G.s_magnetite.

Table S2

Genes predicated to encode proteins that were differentially expressed in G. metallireducens cells grown in both co-cultures of G.m_G.s_GAC and G.m_G.s_magnetite compared with those grown in G.m_G.s. The values were normalised log base2 (ratio: G.m_G.s_GAC/G.m_G.s, G.m_G.s_magnetite/G.m_G.s).

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Supplementary material, approximately 60.6 KB.

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Zheng, S., Liu, F., Li, M. et al. Comparative transcriptomic insights into the mechanisms of electron transfer in Geobacter co-cultures with activated carbon and magnetite. Sci. China Life Sci. 61, 787–798 (2018). https://doi.org/10.1007/s11427-017-9177-1

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