Abstract
Purpose
Legumes form root nodules to gain fixed nitrogen from rhizobia and can also access nitrogen in soil. Data suggest that plants might discriminate among these sources to optimize growth, but recognition of symbiotically fixed nitrogen and its regulation remain poorly understood.
Methods
A greenhouse inoculation study manipulated the molecular form and concentration of nitrogen available using two Lotus japonicus genotypes and the nitrogen-fixing symbiont, Mesorhizobium loti. Plants were supplied with sole organic and inorganic nitrogen sources to simulate forms that plants might receive from symbiotic nitrogen fixation or from the soil. Host benefit from and regulation of symbiosis was investigated by quantifying symbiotic trait variation and isotopic analysis of nitrogen fixation.
Results
Host growth varied in response to fertilization with alanine, aspartic acid, ammonium, and nitrate, suggesting differences in catabolism efficiency. Net benefits of nodulation were reduced or eliminated under all forms of extrinsic fertilization. However, even when symbiosis imposed significant costs, hosts did not reduce investment into nodulation or nitrogen fixation when exposed to aspartic acid, unlike with other nitrogen sources.
Conclusions
L. japonicus can adaptively downregulate investment into symbiosis in the presence of some but not all nitrogen sources. Failure to downregulate any aspect of symbiosis in the presence of aspartic acid suggests that it might be jamming the main signal used by L. japonicus to detect nitrogen fixation.
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References
Appels MA, Haaker H (1991) Glutamate oxaloacetate transaminase in pea root nodules. Plant Physiol 95:740–747
Agren G, Franklin O (2003) Root: shoot ratios, optimization and nitrogen productivity. Ann Bot 92:795–800. https://doi.org/10.1093/aob/mcg203
Amarger N (1981) Competition for nodule formation between effective and ineffective strains of Rhizobium meliloti. Soil Biol Biochem 13:475–480. https://doi.org/10.1016/0038-0717(81)90037-7
Bates D, Mächler M, Bolker B, Walker S (2015) Fitting linear mixed effects models using lme4. J Stat Softw 67:48
Batstone RT, Peters MAE, Simonsen AK, Stinchcombe JR, Frederickson ME (2020) Environmental variation impacts trait expression and selection in the legume-rhizobium symbiosis. Am J Bot 107(2):195–208. https://doi.org/10.1002/ajb2.1432
Bull JJ, Rice WR (1991) Distinguishing mechanism for the evolution of co-operation. J Theor Biol 149(1):63–74. https://doi.org/10.1016/S0022-5193(05)80072-4
Buzas DM, Gresshoff PM (2007) Short and long-distance control of root development by LjHAR1 during the juvenile stage of Lotus japonicus. J Plant Physiol 164:452–459
Carroll BJ, McNeil DL, Gresshoff PM (1985) A supernodulation and nitrate-tolerant symbiotic (nts) soybean mutant. Plant Physiol 78:34–40
Champion RA, Mathis JN, Israel DW, Hunt PG (1992) Response of soybean to inoculation with efficient and inefficient Bradyrhizobium japonicum variants. Crop Sci 32:457–463. https://doi.org/10.2135/cropsci1992.0011183X003200020034x
Corrales A, Mangan SA, Turner BL, Dalling JW (2016) An ectomycorrhizal nitrogen economy facilitates monodominance in a neotropical forest. Ecol Lett 19:383–392. https://doi.org/10.1111/ele.12570
Ferguson BJ, Mens C, Hastwell AH, Zhang M, Su H, Jones CH, Chu X, Gresshoff PM (2018) Legume nodulation: The host controls the party. Plant, Cell Environ 42(1):41–51. https://doi.org/10.1111/pce.13348
Fox J, Weisberg S (2019) An R companion to applied regression. Sage, Thousand Oaks
Galloway JN, Dentener DG, Capone EW, Boyer RW, Howarth S, Seitzinger P, Asner GP, Cleveland CC, Green PA, Holland EA, Karl DM, Michaels AF, Porter JH, Townsend AR, Vorosmarty CJ (2004) Nitrogen cycles: past, present, and future. Biogeochemistry 70(2):153–226. https://doi.org/10.1007/s10533-004-0370-0
Gan Y, Stulen I, Keulen HV, Kuiper PJC (2004) Low concentrations of nitrate and ammonium stimulate nodulation and N2 fixation while inhibiting specific nodulation (nodule DW g-1 root dry weight) and specific N2 fixation (N2 fixed g-1 root dry weight) in soybean. Plant Soil 258:281–292. https://doi.org/10.1023/B:PLSO.0000016558.32575.17
Gano-Cohen KA, Wendlant CE, Moussawi KA, Stokes PJ, Quides KW, Weisberg AJ, Chang JF, Sachs JL (2020) Recurrent mutualism breakdown events in a legume rhizobia metapopulation. Proc R Soc B 287:20192549. https://doi.org/10.1098/rspb.2019.2549
Gerz M, Bueno CG, Ozinga WA, Zobel M, Moora M (2018) Niche determination and expansion of plant species are associated with mycorrhizal symbiosis. J Ecol 106:254–264. https://doi.org/10.1111/1365-2745.12873
Habinshuti SJ, Maseko ST, Dakora FD (2021) Inhibition of N2 fixation by N fertilization of common bean (Phaseolus vulgaris L.) plants grown on fields of farmers in the eastern cape of South Africa, measured Using 15N natural abundance and tissue ureide analysis. Frontiers in Agronomy. 3:692933. https://doi.org/10.3389/fagro.2021.692933
Hahn M, Studer D (1986) Competitiveness of a nif− Bradyrhizobium japonicum mutant against the wild-type strain. FEMS Microbiol Lett 33:143–148. https://doi.org/10.1111/j.1574-6968.1986.tb01228.x
Heath KD (2010) Intergenomic epistasis and coevolutionary constrains in plants and rhizobia. Evolution 64–5:1446–1458. https://doi.org/10.1111/j.1558-5646.2009.00913.x
Heath KD, Stock AJ, Stinchcombe JR (2010) Mutualism variation in the nodulation response to nitrate. J Evol Biol 23(11):2494–2500. https://doi.org/10.1111/j.1420-9101.2010.02092.x
Hoeksema JD, Chaudhary VB, Gehring CA, Johnson NC, Karst J, Koide RT, Pringle A, Zabinski C, Bever JD, Moore JC, Wilson GWT, Klironomos JN, Umbanhowar J (2010) A meta-analysis of context-dependency in plant response to inoculation with mycorrhizal fungi. Ecol Lett 13:394–407. https://doi.org/10.1111/j.1461-0248.2009.01430.x
Hoeksema JD, Bruna EM (2015) Context-dependent Outcomes of Mutualistic Interactions. In: Bronstein JL (ed) Mutualism. Oxford University Press, pp 181–202
Hussain AKM, Jiang Q, Broughton WJ, Gresshoff PM (1999) Lotus japonicus nodulates and fixes nitrogen with the broad host range Rhizobium sp. NGR234. Plant Cell Physiol 40(8):894–899. https://doi.org/10.1093/oxfordjournals.pcp.a029619
Kawaguchi M (2000) Lotus japonicus ‘Miyakojima’ MG-20: An early-flowering accession suitable for indoor handling. J Plant Res 113:507–509. https://doi.org/10.1007/PL00013961
Kawaguchi M, Imaizumi-Anraku H, Koiwa H, Niwa S, Ikuta A, Syono K, Akao S (2002) Root, root hair, and symbiotic mutants of the model legume Lotus japonicus. Molec Plant Microbe Interact 15(1):17–26
Kiers T, Rousseau RA, West SA, Denison RF (2003) Host Sanctions and the legume-rhizobium mutualism. Nature 425:78–81. https://doi.org/10.1038/nature01931
Kiers ET, Rousseau RA, Denison RF (2006) Measured sanctions: legumes hosts detect quantitative variation in rhizobium cooperation and punish accordingly. Evol Ecol Res 8:1077–1086
Klucas RV (1974) Studies on soybean nodule senescence. Plant Physiol 54:612–616
Krusell L, Madsen LH, Sato S, Aubert G, Genua A, Szczyglowski K, Duc G, Kaneko T, Tabata S, de Bruijn F, Pajuelo E, Sandal N, Stougaard J (2002) Shoot control of root development and nodulation is mediated by a receptor-like kinase. Nature 420:422–426. https://doi.org/10.1038/nature01207
Lenth, R. V. (2016). Least-Squares Means: The R Package lsmeans. J Stat Software, 69(1):1–33. https://doi.org/10.18637/jss.v069.i01
Lin J, Roswanjaya YP, Kohlen W, Stougaard J, Reid D (2021) Nitrate restricts nodule organogenesis through inhibition of cytokinin biosynthesis in Lotus japonicus. Nat Commun 12:6544. https://doi.org/10.1038/s41467-021-26820-9
Lodwig EM, Hosie AHF, Bourdès A, Findlay K, Allaway D, Karunakaran R, Downie DA, Poole PS (2003) Amino-acid cycling drives nitrogen fixation in the legume-Rhizobium symbiosis. Nature 422:722–726. https://doi.org/10.1038/nature01527
Magori S, Oka-Kira E, Shibata S, Umehara Y, Kouchi H, Hase Y, Tanaka A, Sato S, Tabata S, Kawaguchi M (2009) TOO MUCH LOVE, a root regulator associated with the long-distance control of nodulation in Lotus japonicus. Molec Plant Microbe Interact 22(3):259–268. https://doi.org/10.1094/MPMI-22-3-0259
Meeks JC, Wolk CP, Schilling N, Shaffer PW (1978) Initial organic products of fixation of [13N]dinitrogen by root nodules of soybean (Glycine max). Plant Physiol 61:980–983
Murray-Stoker D, Johnson MTJ (2021) Ecological consequences of urbanization on a legume-rhizobia mutualism. Oikos 130:1750–1761. https://doi.org/10.1111/oik.08341
Ngom M, Gray K, Diagne N, Oshone R, Fardoux J, Gherbi H, Hocher V, Svistoonoff S, Laplaze L, Tisa LS, Sy MO, Champion A (2016) Symbiotic performance of diverse Frankia strains on salt-stressed Casuarina glauca and Casuarina equisetifolia plants. Front Plant Sci 7:1331. https://doi.org/10.3389/fpls.2016.01331
Nishimura R, Hayashi M, Wu G, Kouchi H, Imaizumi-Anraku H, Murakami Y, Kawasaki S, Akao S, Ohmori M, Nagasawa M, Harada K, Kawaguchi M (2002) HAR1 mediates systemic regulation of symbiotic organ development. Nature 420:426–429. https://doi.org/10.1038/nature01231
Ohyama T, Kumazawa K (1980) Nitrogen assimilation in soybean nodules. Soil Sci Plant Nutr 26(2):205–213
Oka-Kira E, Tateno K, Miura K, Haga T, Hayashi M, Harada K, Sato S, Tabata S, Shikazono N, Tanaka A, Watanabe Y, Fukuhara I, Nagata T, Kawaguchi M (2005) Klavier (klv), a novel hypernodulation mutant of Lotus japonicus affected in vascular tissue organization and floral induction. Plant J 44:505–515. https://doi.org/10.1111/j.1365-313X.2005.02543.x
Oono R, Anderson CG, Denison RF (2011) Failure to fix nitrogen by non-reproductive symbiotic rhizobia triggers host sanctions that reduce fitness of their reproductive clonemates. Proc R Soc 278:2698–2705. https://doi.org/10.1098/rspb.2010.2193
Ortiz-Barbosa GS, Torres-Martinez L, Manci A, Neal S, Soubra T, Khairi F, Trinh J, Cardenas P, Sachs JL (2022) No disruption of rhizobial symbiosis during early stages of cowpea domestication. Evolution 76(3):496–511. https://doi.org/10.1111/evo.14424
Pfau T, Christian N, Masakapalli SK, Sweetlove LJ, Poolman MG, Ebenhoh O (2018) The interwined metabolism during symbiotic nitrogen fixation elucidated by metabolic modelling. Sci Rep 8:12504. https://doi.org/10.1038/s41598-018-30884-x
Porter SS, Simms EL (2014) Selection for cheating across disparate environments in the legume-rhizobium mutualism. Ecol Lett 17:1121–1129. https://doi.org/10.1111/ele.12318
Quides KW, Stomackin GM, Lee H, Chang JH, Sachs JL (2017) Lotus japonicus alters in planta fitness of Mesorhizobium loti dependent on symbiotic nitrogen fixation. PLoS ONE. 12(9):e0185568. https://doi.org/10.1371/journal.pone.0185568
Quides KW, Salaheldine F, Jariwala R, Sachs JL (2021) Dysregulation of host-control causes interspecific conflict over host investment into symbiotic organs. Evolution. https://doi.org/10.1111/evo.14173
Rastogi VP, Watson RJ (1991) Aspartate aminotransferase activity is required for aspartate catabolism and symbiotic nitrogen fixation in Rhizobium meliloti. J Bacteriol 2879–2887. https://doi.org/10.1128/jb.173.9.2879-2887.1991
Regus JU, Gano KA, Hollowell AC, Sachs JL (2014) Efficiency of partner choice and sanctions in Lotus is not altered by nitrogen fertilization. Proc R Soc B 281:20132587. https://doi.org/10.1098/rspb.2013.2587
Regus JU, Gano KA, Hollowell AC, Sofish V, Sachs JL (2015) Lotus hosts delimit the mutualism-parasitism continuum of Bradyrhizobium. J Evol Biol 28:447–456. https://doi.org/10.1111/jeb.12579
Regus JU, Wendlandt CE, Bantay RM, Gano-Cohen KA, Gleason NJ, Hollowell AC, O’Neill MR, Shahin KK, Sachs JL (2017a) Nitrogen deposition decreases the benefits of symbiosis in a native legume. Plant Soil 2017(414):159–170. https://doi.org/10.1007/s11104-016-3114-8
Regus JU, Quides KW, O’Neill MR, Suzuki R, Savory EA, Chang JH, Sachs JL (2017b) Cell autonomous sanctions in legumes target ineffective rhizobia in nodules with mixed infections. Am J Bot 104(9):1–14. https://doi.org/10.3732/ajb.1700165
Reinprecht Y, Schram L, Marsolais F, Smith TH, Hill B, Pauls KP (2020) Effects of nitrogen application on nitrogen fixation in common bean production. Front Plant Sci 11:1172. https://doi.org/10.3389/fpls.2020.01172
Rosendahl L, Dilworth MJ, Glenn AR (1992) Exchange of metabolites across the peribacteroid membrane in pea root nodules. J Plant Physiol. 139: 635–638.
Sachs JL, Simms EL (2008) The origins of uncooperative rhizobia. Oikos 117:961–964. https://doi.org/10.1111/j.2008.0030-1299.16606.x
Sachs JL, Kembel SW, Lau AH, Simms EL (2009) In situ phylogenetic structure and diversity of wild Bradyrhizobium communities. Appl Environ Microbiol 75:4727–4735. https://doi.org/10.1128/AEM.00667-09
Sachs JL, Russel JE, Lii YE, Black KC, Lopez G, Patil AS (2010) Host control over infection and proliferation of a cheater symbiont. J Evol Biol 23:1919–1927. https://doi.org/10.1111/j.1420-9101.2010.02056.x
Sachs JL, Quides KW, Wendlant CE (2018) Legumes versus rhizobia: a model for ongoing conflict in symbiosis. New Phytol 219:1199–1206. https://doi.org/10.1111/nph.15222
Sawada H, Kuykendall LD, Young JM (2003) Changing concepts in the systematics of bacterial nitrogen-fixing legume symbionts. J Gen Appl Microbiol 49:155–179. https://doi.org/10.2323/jgam.49.155
Schauser L, Handberg K, Sandal N, Stiller J, Thykjaer T, Pajuelo E, Nielsen A, Stougaard J (1998) Symbiotic mutants deficient in nodule establishment identified after T-DNA transformation of Lotus japonicus. Mol Gen Genet 159:414–423
Schnabel E, Journet EP, de Carvalho-Niebel F, Duc G, Frugoli J (2005) The Medicago truncatula SUNN gene encodes a CLV1-like leucine-rich repeat receptor kinase that regulates nodule number and root length. Plant Mol Biol 58:809–822
Simard SW, Beiler KJ, Bingham MA, Deslippe JR, Philip LJ, Teste FP (2012) Mycorrhizal networks: Mechanisms, ecology and modelling. Fungal Biol Rev 39–60. https://doi.org/10.1016/j.fbr.2012.01.001
Simonsen AK, Han S, Rekret P, Rentschler CS, Heath KD, Stinchcombe JR (2015) Short-term fertilizer application alters phenotypic traits of symbiotic nitrogen fixing bacteria. PeerJ 3:e1291. https://doi.org/10.7717/peerj.1291
Simonsen AK, Dinnage R, Barrett LG, Prober SM, Thrall PH (2017) Symbiosis limits establishment of legumes outside their native range at a global scale. Nat Commun 8:14790. https://doi.org/10.1038/ncomms14790
Somasegaran P, Hoben HJ (1994) Handbook for rhizobia: methods in legume rhizobium technology. Springer Verlag, New York, p 450
Streeter J (1988) Inhibition of legume nodule formation and N2 fixation by nitrate. Crit Rev Plant Sci 7(1):1–23. https://doi.org/10.1080/07352688809382257
Sulieman S, Tran LP (2012) Asparagine: an amide of particular disctinction in the regulation of symbiotic nitrogen fixation of legumes. Crit Rev Biotechnol 1–19. https://doi.org/10.3109/07388551.2012.695770
Szczyglowski K, Shaw RS, Wopereis J, Copeland S, Hamburger D, Kasiborski B, Dazzo FB, de Bruijn FJ (1998) Nodule organogenesis and symbiotic mutants of the model legume Lotus japonicus. Molecular Plant Microbe Interactions 11(7):684–697
Tyerman SD, Whitehead LF, Day DA (1995) A channel-like transporter for NH4+ on the symbiotic interface of N2-fixing plants. Nature 378:629–632
Unkovich M, Herridge D, Peoples M, Cadisch G, Boddey B, Giller K, Alves B, Chalk P (2008) Measuring Plant-associated Nitrogen Fixation in Agricultural Systems. Australian Centre for International Agricultural Research (ACIAR), Canberra
Waters JK, Hughes BL II, Purcell LC, Gerhardt KO, Mawhinney TP, Emerich DW (1998) Alanine, not ammonia, is excreted from N2-fixing soybean nodule bacterioids. Proc Natl Acad Sci 95:12038–12042. https://doi.org/10.1073/pnas.95.20.12038
Weese DJ, Heath KD, Dentinger BTM, Lau JA (2015) Long-term nitrogen addition causes the evolution of less cooperative mutualists. Evolution 69(3):631–642. https://doi.org/10.1111/evo.12594
Wendlandt CE, Regus JU, Gano-Cohen KA, Hollowell AC, Quides KW, Lyu JY, Adinata ES, Sachs JL (2019) Host investment into symbiosis varies among genotypes of the legume Acmispon strigosus, but host sanctions are uniform. New Phytol 221:446–458. https://doi.org/10.1111/nph.15378
West SA, Kiers ET, Simms EL, Denison RF (2002) Sanctions and mutualism stability: why do rhizobia fix nitrogen. Proc R Soc London B 269:685–694. https://doi.org/10.1098/rspb.2001.1878
Westhoek A, Field E, Rehling F, Mulley G, Webb I, Poole PS, Turnbull LA (2017) Policing the legume-Rhizobium symbiosis: a critical test of partner choice. Sci Rep 7:1419. https://doi.org/10.1038/s41598-017-01634-2
White J, Prell J, James EK, Poole P (2007) Nutrient sharing between symbionts. Plant Physiol 144:604–614. https://doi.org/10.1104/pp.107.097741
Wopereis J, Pajuelo E, Dazzo FB, Jiang Q, Gresshoff PM, De Bruijn FJ, Stougaard J, Szczyglowski K (2000) Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype. Plant J 23(1):97–114
Xia X, Ma C, Dong S, Xu Y, Gong Z (2017) Effects of nitrogen concentrations on nodulation and nitrogenase activity in dual root systems of soybean plants. Soil Sci Plant Nutr 63(5):470–482. https://doi.org/10.1080/00380768.2017.1370960
Acknowledgements
We thank LegumeBase at the University of Miyazaki Japan, from which we acquired MG-20 seeds. We thank Masayoshi Kawaguchi at the National Institute for Basic Biology, Okazaki, Aichi, Japan, for providing the har1-7 mutant seeds.
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11104_2022_5762_MOESM1_ESM.csv
Supplementary file1 Experimental setup and plants during harvest. (A) Experimental set up in the greenhouse (B) har1-7 plant inoculated with rhizobia and fertilized with potassium nitrate (100%) (C) MG20 plant inoculated with rhizobia and fertilized with potassium nitrate (100%). (CSV 76.0 KB)
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Ortiz-Barbosa, G.S., Torres-Martínez, L., Rothschild, J. et al. Lotus japonicus regulates root nodulation and nitrogen fixation dependent on the molecular form of nitrogen fertilizer. Plant Soil 483, 533–545 (2023). https://doi.org/10.1007/s11104-022-05762-1
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DOI: https://doi.org/10.1007/s11104-022-05762-1