Abstract
The largest natural source of methane (CH4) to the atmosphere is wetlands, which produce 20% to 50% of total global emissions. Vascular plants play a key role regulating wetland CH4 emissions through multiple mechanisms. They often contain aerenchymatous tissues which act as a diffusive pathway for CH4 to travel from the anoxic soil to the atmosphere and for O2 to diffuse into the soil and enable methanotrophy. Plants also exude carbon from their roots which stimulates microbial activity and fuels methanogenesis. This study investigated these mechanisms in a laboratory experiment utilizing rootboxes containing either Carex aquatilis plants, silicone tubes that simulated aerenchymatous gas transfer, or only soil as a control. CH4 emissions were over 50 times greater from planted boxes than from control boxes or simulated plants, indicating that the physical transport pathway of aerenchyma was of little importance when not paired with other effects of plant biology. Plants were exposed to 13CO2 at two time-points and subsequent enrichment of root tissue, rhizosphere soil, and emitted CH4 was used in an isotope mixing model to determine the proportion of plant-derived versus soil-derived carbon supporting methanogenesis. Results showed that carbon exuded by plants was converted to CH4 but also that planted boxes emitted 28 times more soil-derived carbon than the other experimental treatments. At the end of the experiment, emissions of excess soil-derived carbon from planted boxes exceeded the emission of plant-derived carbon. This result signifies that plants and root exudates altered the soil chemical environment, increased microbial metabolism, and/or changed the microbial community such that microbial utilization of soil carbon was increased (e.g. microbial priming) and/or oxidation of soil-derived CH4 was decreased (e.g., by microbial competition for oxygen).
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References
Aller RC (1998) Mobile deltaic and continental shelf muds as suboxic, fluidized bed reactors. Mar Chem 61:143–155. https://doi.org/10.1016/s0304-4203(98)00024-3
Armstrong W (1971) Radial oxygen losses from intact rice roots as affected by distance from the apex, respiration and waterlogging. Physiol Plant 25:192–197. https://doi.org/10.1111/j.1399-3054.1971.tb01427.x
Aselmann I, Crutzen PJ (1989) Global distribution of natural freshwater wetlands and rice paddies, their net primary productivity, seasonality and possible methane emissions. J Atmos Chem 8:307–358. https://doi.org/10.1007/bf00052709
Basiliko N, Stewart H, Roulet NT, Moore TR (2012) Do root exudates enhance peat decomposition? Geomicrobiol J 29:374–378. https://doi.org/10.1080/01490451.2011.568272
Bender MM (1971) Variations in the 13C/12C ratios of plants in relation to the pathway of photosynthetic carbon dioxide fixation. Phytochemistry 10:1239–1244
Botz R, Pokojski H-D, Schmitt M, Thomm M (1996) Carbon isotope fractionation during bacterial methanogenesis by CO2 reduction. Org Geochem 25:255–262. https://doi.org/10.1016/s0146-6380(96)00129-5
Canfield DE (1994) Factors influencing organic carbon preservation in marine sediments. Chem Geol 114:315–329. https://doi.org/10.1016/0009-2541(94)90061-2
Carvalhais LC, Dennis PG, Fedoseyenko D, Hajirezaei M-R, Borriss R, von Wirén N (2011) Root exudation of sugars, amino acids, and organic acids by maize as affected by nitrogen, phosphorus, potassium, and iron deficiency. J Plant Nutr Soil Sci 174:3–11. https://doi.org/10.1002/jpln.201000085
Chanton JP, Martens CS, Kelley CA, Crill PM, Showers WJ (1992) Methane transport mechanisms and isotopic fractionation in emergent macrophytes of an Alaskan tundra lake. J Geophys Res. https://doi.org/10.1029/90jd01542
Chanton JP, Glaser PH, Chasar LS, Burdige DJ, Hines ME, Siegel DI, Tremblay LB, Cooper WT (2008) Radiocarbon evidence for the importance of surface vegetation on fermentation and methanogenesis in contrasting types of boreal peatlands. Glob Biogeochem Cycles. https://doi.org/10.1029/2008gb003274
Chen R, Senbayram M, Blagodatsky S, Myachina O, Dittert K, Lin X, Blagodatskaya E, Kuzyakov Y (2014) Soil C and N availability determine the priming effect: microbial N mining and stoichiometric decomposition theories. Glob Change Biol 20:2356–2367. https://doi.org/10.1111/gcb.12475
Ciais P, Sabine C, Bala G, Bopp L, Brovkin V, Canadell J, Chhabra A, DeFries R, Galloway J, Heimann M, Jones C, Le Quéré C, Myneni RB, Piao S, Thornton P (2013) Carbon and other biogeochemical cycles. In: Stocker TF, Qin D, Plattner G-K, Tignor M, Allen SK, Boschung J, Nauels A, Xia Y, Bex V, Midgley PM (eds) Climate change 2013: the physical science basis. Contribution of Working Group I to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press, Cambridge, pp 465–570
Conrad R (2009) The global methane cycle: recent advances in understanding the microbial processes involved. Environ Microbiol Rep 1:285–292. https://doi.org/10.1111/j.1758-2229.2009.00038.x
Coplen TB, Brand WA, Gehre M, Gröning M, Meijer HAJ, Toman B, Verkouteren RM (2006) New guidelines for δ13C measurements. Anal Chem 78:2439–2441. https://doi.org/10.1021/ac052027c
Corbett JE, Tfaily MM, Burdige DJ, Glaser PH, Chanton JP (2015) The relative importance of methanogenesis in the decomposition of organic matter in northern peatlands. J Geophys Res Biogeosci 120:280–293. https://doi.org/10.1002/2014jg002797
Craine JM, Morrow C, Fierer N (2007) Microbial nitrogen limitation increases decomposition. Ecology 88:2105–2113. https://doi.org/10.1890/06-1847.1
Crombie AT, Murrell JC (2014) Trace-gas metabolic versatility of the facultative methanotroph Methylocella silvestris. Nature 510:148–151. https://doi.org/10.1038/nature13192
Dedysh SN, Knief C, Dunfield PF (2005) Methylocella species are facultatively methanotrophic. J Bacteriol 187:4665–4670. https://doi.org/10.1128/jb.187.13.4665-4670.2005
Ding W, Cai Z, Tsuruta H (2004) Summertime variation of methane oxidation in the rhizosphere of a Carex dominated freshwater marsh. Atmos Environ 38:4165–4173. https://doi.org/10.1016/j.atmosenv.2004.04.022
Dorodnikov M, Knorr K-H, Kuzyakov Y, Wilmking M (2011) Plant-mediated CH4 transport and contribution of photosynthates to methanogenesis at a boreal mire: a 14C pulse-labeling study. Biogeosciences 8:2365–2375. https://doi.org/10.5194/bg-8-2365-2011
Feisthauer S, Vogt C, Modrzynski J, Szlenkier M, Krüger M, Siegert M, Richnow H-H (2011) Different types of methane monooxygenases produce similar carbon and hydrogen isotope fractionation patterns during methane oxidation. Geochim Cosmochim Acta 75:1173–1184. https://doi.org/10.1016/j.gca.2010.12.006
Fetzer S, Conrad R (1993) Effect of redox potential on methanogenesis by Methanosarcina barkeri. Arch Microbiol 160:108–113. https://doi.org/10.1007/bf00288711
Forkel M, Carvalhais N, Rödenbeck C, Keeling R, Heimann M, Thonicke K, Zaehle S, Reichstein M (2016) Enhanced seasonal CO2 exchange caused by amplified plant productivity in northern ecosystems. Science 351:696–699. https://doi.org/10.1126/science.aac4971
Fritz C, Pancotto VA, Elzenga JTM, Visser EJW, Grootjans AP, Pol A, Iturraspe R, Roelofs JGM, Smolders AJP (2011) Zero methane emission bogs: extreme rhizosphere oxygenation by cushion plants in Patagonia. N Phytol 190:398–408. https://doi.org/10.1111/j.1469-8137.2010.03604.x
Galand P, Yrjälä K, Conrad R (2010) Stable carbon isotope fractionation during methanogenesis in three boreal peatland ecosystems. Biogeosciences. https://doi.org/10.5194/bg-7-3893-2010
Games LM, HayesRobert JM, Gunsalus P (1978) Methane-producing bacteria: natural fractionations of the stable carbon isotopes. Geochim Cosmochim Acta 42:1295–1297. https://doi.org/10.1016/0016-7037(78)90123-0
Gaudinski JB, Trumbore SE, Davidson EA, Zheng S (2000) Soil carbon cycling in a temperate forest: radiocarbon-based estimates of residence times, sequestration rates and partitioning of fluxes. Biogeochemistry 51:33–69. https://doi.org/10.1023/a:1006301010014
Gedney N, Cox PM, Huntingford C (2004) Climate feedback from wetland methane emissions. Geophys Res Lett 31:L20503. https://doi.org/10.1029/2004gl020919
Gelwicks JT, Risatti JB, Hayes JM (1994) Carbon isotope effects associated with aceticlastic methanogenesis. Appl Environ Microbiol 60:467–472
Girkin NT, Turner BL, Ostle N, Craigon J, Sjögersten S (2018a) Root exudate analogues accelerate CO2 and CH4 production in tropical peat. Soil Biol Biochem 117:48–55. https://doi.org/10.1016/j.soilbio.2017.11.008
Girkin NT, Turner BL, Ostle N, Sjögersten S (2018b) Composition and concentration of root exudate analogues regulate greenhouse gas fluxes from tropical peat. Soil Biol Biochem 127:280–285. https://doi.org/10.1016/j.soilbio.2018.09.033
Graham EB, Tfaily MM, Crump AR, Goldman AE, Bramer LM, Arntzen E, Romero E, Resch CT, Kennedy DW, Stegen JC (2017) Carbon inputs from riparian vegetation limit oxidation of physically bound organic carbon via biochemical and thermodynamic processes. J Geophys Res Biogeosci 122:3188–3205. https://doi.org/10.1002/2017jg003967
Hamer U, Marschner B (2002) Priming effects of sugars, amino acids, organic acids and catechol on the mineralization of lignin and peat. J Plant Nutr Soil Sci 165:261–268. https://doi.org/10.1002/1522-2624(200206)165:3%3c261:aid-jpln261%3e3.0.co;2-i
Hodson EL, Poulter B, Zimmermann NE, Prigent C, Kaplan JO (2011) The El Niño-Southern Oscillation and wetland methane interannual variability. Geophys Res Lett 38:L08810. https://doi.org/10.1029/2011gl046861
Houlden A, Timms-Wilson TM, Day MJ, Bailey MJ (2008) Influence of plant developmental stage on microbial community structure and activity in the rhizosphere of three field crops. FEMS Microbiol Ecol 65:193–201. https://doi.org/10.1111/j.1574-6941.2008.00535.x
Idso SB, Kimball BA, Anderson MG, Mauney JR (1987) Effects of atmospheric CO2 enrichment on plant growth: the interactive role of air temperature. Agric Ecosyst Environ 20:1–10. https://doi.org/10.1016/0167-8809(87)90023-5
Intergovernmental Panel on Climate Change (ed) (2014) Climate change 2013—the physical science basis: Working Group I Contribution to the Fifth Assessment Report of the Intergovernmental Panel on Climate Change. Cambridge University Press, Cambridge
Joabsson A, Christensen TR, Wallén B (1999) Vascular plant controls on methane emissions from northern peatforming wetlands. Trends Ecol Evol 14:385–388. https://doi.org/10.1016/s0169-5347(99)01649-3
Jonasson S, Lee JA, Callaghan TV, Havström M, Parsons AN (1996) Direct and indirect effects of increasing temperatures on subarctic ecosystems. Ecol Bull 180–191
Jones DL, Dennis PG, Owen AG, van Hees PAW (2003) Organic acid behavior in soils—misconceptions and knowledge gaps. Plant Soil 248:31–41. https://doi.org/10.1023/a:1022304332313
Kayranli B, Scholz M, Mustafa A, Hedmark Å (2009) Carbon storage and fluxes within freshwater wetlands: a critical review. Wetlands 30:111–124. https://doi.org/10.1007/s13157-009-0003-4
Keiluweit M, Nico PS, Kleber M, Fendorf S (2016) Are oxygen limitations under recognized regulators of organic carbon turnover in upland soils? Biogeochemistry 127:157–171. https://doi.org/10.1007/s10533-015-0180-6
King JY, Reeburgh WS, Regli SK (1998) Methane emission and transport by Arctic sedges in Alaska: results of a vegetation removal experiment. J Geophys Res Atmos 103:29083–29092. https://doi.org/10.1029/98jd00052
Krohn J, Lozanovska I, Kuzyakov Y, Parvin S, Dorodnikov M (2017) CH4 and CO2 production below two contrasting peatland micro-relief forms: an inhibitor and δ13C study. Sci Total Environ 586:142–151. https://doi.org/10.1016/j.scitotenv.2017.01.192
Kummerow J, Ellis BA (1984) Temperature effect on biomass production and root/shoot biomass ratios in two Arctic sedges under controlled environmental conditions. Can J Bot 62:2150–2153. https://doi.org/10.1139/b84-294
Kuzyakov Y (2010) Priming effects: interactions between living and dead organic matter. Soil Biol Biochem 42:1363–1371. https://doi.org/10.1016/j.soilbio.2010.04.003
Larsen M, Borisov SM, Grunwald B, Klimant I, Glud RN (2011) A simple and inexpensive high resolution color ratiometric planar optode imaging approach: application to oxygen and pH sensing. Limnol Oceanogr Methods 9:348–360. https://doi.org/10.4319/lom.2011.9.348
Lehmeier CA, Ballantyne F, Min K, Billings SA (2016) Temperature-mediated changes in microbial carbon use efficiency and 13C discrimination. https://doi.org/10.5194/bg-13-3319-2016
Lenzewski N, Mueller P, Meier RJ, Liebsch G, Jensen K, Koop-Jakobsen K (2018) Dynamics of oxygen and carbon dioxide in rhizospheres of Lobelia dortmanna—a planar optode study of belowground gas exchange between plants and sediment. N Phytol 218:131–141. https://doi.org/10.1111/nph.14973
Londry KL, Dawson KG, Grover HD, Summons RE, Bradley AS (2008) Stable carbon isotope fractionation between substrates and products of Methanosarcina barkeri. Org Geochem 39:608–621. https://doi.org/10.1016/j.orggeochem.2008.03.002
Lu Y, Conrad R (2005) In situ stable isotope probing of methanogenic archaea in the rice rhizosphere. Science 309:1088–1090. https://doi.org/10.1126/science.1113435
Manies KL, Fuller CC, Jones MC, Waldrop MP, McGeehin JP (2017) Soil data for a thermokarst bog and the surrounding permafrost plateau forest, located at Bonanza Creek Long Term Ecological Research Site, Interior Alaska. U.S. Geological Survey, Reston
Megonigal JP, Schlesinger WH (1997) Enhanced CH4 emission from a wetland soil exposed to elevated CO2. Biogeochemistry 37:77–88. https://doi.org/10.1023/a:1005738102545
Megonigal JP, Whalen SC, Tissue DT, Bovard BD, Allen AS, Albert DB (1999) A plant–soil–atmosphere microcosm for tracing radiocarbon from photosynthesis through methanogenesis. Soil Sci Soc Am J 63:665–671. https://doi.org/10.2136/sssaj1999.03615995006300030033x
Mueller P, Jensen K, Megonigal JP (2016) Plants mediate soil organic matter decomposition in response to sea level rise. Glob Change Biol 22:404–414. https://doi.org/10.1111/gcb.13082
Neumann RB, Blazewicz SJ, Conaway CH, Turetsky MR, Waldrop MP (2015) Modeling CH4 and CO2 cycling using porewater stable isotopes in a thermokarst bog in Interior Alaska: results from three conceptual reaction networks. Biogeochemistry 127:57–87. https://doi.org/10.1007/s10533-015-0168-2
Nielsen CS, Michelsen A, Ambus P, Deepagoda TKKC, Elberling B (2017) Linking rhizospheric CH4 oxidation and net CH4 emissions in an arctic wetland based on 13CH4 labeling of mesocosms. Plant Soil 412:201–213. https://doi.org/10.1007/s11104-016-3061-4
O’Leary MH (1988) Carbon isotopes in photosynthesis. BioScience 38:328–336. https://doi.org/10.2307/1310735
Picek T, Čížková H, Dušek J (2007) Greenhouse gas emissions from a constructed wetland—plants as important sources of carbon. Ecol Eng 31:98–106. https://doi.org/10.1016/j.ecoleng.2007.06.008
Popp TJ, Chanton JP, Whiting GJ, Grant N (1999) Methane stable isotope distribution at a Carex dominated fen in north central Alberta. Glob Biogeochem Cycles 13:1063–1077. https://doi.org/10.1029/1999gb900060
Popp TJ, Chanton JP, Whiting GJ, Grant N (2000) Evaluation of methane oxidation in the rhizosphere of a Carex dominated fen in northcentral Alberta, Canada. Biogeochemistry 51:259–281. https://doi.org/10.1023/a:1006452609284
Rasse DP, Rumpel C, Dignac M-F (2005) Is soil carbon mostly root carbon? Mechanisms for a specific stabilisation. Plant Soil 269:341–356. https://doi.org/10.1007/s11104-004-0907-y
Riley WJ, Subin ZM, Lawrence DM, Swenson SC, Torn MS, Meng L, Mahowald NM, Hess P (2011) Barriers to predicting changes in global terrestrial methane fluxes: analyses using CLM4Me, a methane biogeochemistry model integrated in CESM. Biogeosciences 8:1925–1953. https://doi.org/10.5194/bg-8-1925-2011
Robroek BJM, Albrecht RJH, Hamard S, Pulgarin A, Bragazza L, Buttler A, Jassey VE (2016) Peatland vascular plant functional types affect dissolved organic matter chemistry. Plant Soil 407:135–143. https://doi.org/10.1007/s11104-015-2710-3
Schimel JP (1995) Plant transport and methane production as controls on methane flux from Arctic wet meadow tundra. Biogeochemistry 28:183–200. https://doi.org/10.1007/bf02186458
Schipper LA, Reddy KR (1996) Determination of methane oxidation in the rhizosphere of Sagittaria lancifolia using methyl fluoride. Soil Sci Soc Am J 60:611–616. https://doi.org/10.2136/sssaj1996.03615995006000020039x
Segers R, Leffelaar PA (2001) Modeling methane fluxes in wetlands with gas-transporting plants: 1. Single-root scale. J Geophys Res Atmos 106:3511–3528. https://doi.org/10.1029/2000jd900484
Shannon RD, White JR (1994) A three-year study of controls on methane emissions from two Michigan peatlands. Biogeochemistry 27:35–60. https://doi.org/10.1007/bf00002570
Ström L, Christensen TR (2007) Below ground carbon turnover and greenhouse gas exchanges in a sub-Arctic wetland. Soil Biol Biochem 39:1689–1698. https://doi.org/10.1016/j.soilbio.2007.01.019
Ström L, Ekberg A, Mastepanov M, Christensen TR (2003) The effect of vascular plants on carbon turnover and methane emissions from a tundra wetland. Glob Change Biol 9:1185–1192. https://doi.org/10.1046/j.1365-2486.2003.00655.x
Theisen AR, Ali MH, Radajewski S, Dumont MG, Dunfield PF, McDonald IR, Dedysh SN, Miguez CB, Murrell JC (2005) Regulation of methane oxidation in the facultative methanotroph Methylocella silvestris BL2. Mol Microbiol 58:682–692. https://doi.org/10.1111/j.1365-2958.2005.04861.x
Trinder CJ, Artz RRE, Johnson D (2008) Contribution of plant photosynthate to soil respiration and dissolved organic carbon in a naturally recolonising cutover peatland. Soil Biol Biochem 40:1622–1628. https://doi.org/10.1016/j.soilbio.2008.01.016
Turner J, Moorberg CJ, Wong A, Waldo NB, Hunt BK, Shea K, Waldrop MP, Turetsky MR, Neumann RB Getting to the root of plant-mediated methane emissions and oxidation in a thermokarst bog. (Manuscript in review)
Valentine DL, Chidthaisong A, Rice A, Reeburgh WS, Tyler SC (2004) Carbon and hydrogen isotope fractionation by moderately thermophilic methanogens. Associate editor: N. E. Ostrom. Geochim Cosmochim Acta 68:1571–1590. https://doi.org/10.1016/j.gca.2003.10.012
Wang Z-P, Han X-G (2005) Diurnal variation in methane emissions in relation to plants and environmental variables in the Inner Mongolia marshes. Atmos Environ 39:6295–6305. https://doi.org/10.1016/j.atmosenv.2005.07.010
Weigel HJ, Pacholski A, Burkart S, Helal M, Heinemeyer O, Kleikamp B, Manderscheid R, Fruhauf C, Hendrey GF, Lewin K, Nagy J (2005) Carbon turnover in a crop rotation under free air CO2 enrichment (FACE). Pedosphere 15:728–738
Whalen SC (2005) Biogeochemistry of methane exchange between natural wetlands and the atmosphere. Environ Eng Sci 22:73–94. https://doi.org/10.1089/ees.2005.22.73
Whiticar MJ (1999) Carbon and hydrogen isotope systematics of bacterial formation and oxidation of methane. Chem Geol 161:291–314. https://doi.org/10.1016/s0009-2541(99)00092-3
Whiting GJ, Chanton JP (1992) Plant-dependent CH4 emission in a subarctic Canadian fen. Glob Biogeochem Cycles 6:225–231. https://doi.org/10.1029/92gb00710
Wieczorek AS, Drake HL, Kolb S (2011) Organic acids and ethanol inhibit the oxidation of methane by mire methanotrophs. FEMS Microbiol Ecol 77:28–39. https://doi.org/10.1111/j.1574-6941.2011.01080.x
Ye R, Doane TA, Morris J, Horwath WR (2015) The effect of rice straw on the priming of soil organic matter and methane production in peat soils. Soil Biol Biochem 81:98–107. https://doi.org/10.1016/j.soilbio.2014.11.007
Yu ZC (2012) Northern peatland carbon stocks and dynamics: a review. Biogeosciences 9:4071–4085. https://doi.org/10.5194/bg-9-4071-2012
Yvon-Durocher G, Allen AP, Bastviken D, Conrad R, Gudasz C, St-Pierre A, Thanh-Duc N, del Giorgio PA (2014) Methane fluxes show consistent temperature dependence across microbial to ecosystem scales. Nature 507:488–491. https://doi.org/10.1038/nature13164
Zhang Z, Zimmermann NE, Stenke A, Li X, Hodson EL, Zhu G, Huang C, Poulter B (2017) Emerging role of wetland methane emissions in driving 21st century climate change. Proc Natl Acad Sci USA 114:9647–9652. https://doi.org/10.1073/pnas.1618765114
Acknowledgements
We thank Jesse Turner, Megan Nims, Olivia Hargrave, Robert Ardissono and Marina Kochuten for laboratory assistance, Kathryn Cogurt for feedback and editing the manuscript, and Christopher Anderton for coordinating access to instruments at the Environmental Molecular Sciences Laboratory. This material is based upon work supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research under Award Number DE-SC-0010338. A portion of this research was performed under the Facilities Integrating Collaborations for User Science (FICUS) Program and used resources at the Environmental Molecular Sciences Laboratory (grid.436923.9), which is a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research and operated under Contract No. DE-AC05-76RL01830. This material is based upon work supported by the U.S. Department of Energy, Office of Science, Office of Workforce Development for Teachers and Scientists, Office of Science Graduate Student Research (SCGSR) Program. The SCGSR Program is administered by the Oak Ridge Institute for Science and Education (ORISE) for the DOE. ORISE is managed by ORAU under Contract Number DE-SC0014664. Students were additionally supported by the following Fellowships and Grants: UW College Of Engineering Dean’s Fellowship/Ford Motor Company Fellowship, UW CEE Valle Scholarship, UW Mary Gates Scholarship, and the Carleton College Kolenkow Reitz Fellowship.
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Waldo, N.B., Hunt, B.K., Fadely, E.C. et al. Plant root exudates increase methane emissions through direct and indirect pathways. Biogeochemistry 145, 213–234 (2019). https://doi.org/10.1007/s10533-019-00600-6
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DOI: https://doi.org/10.1007/s10533-019-00600-6