Abstract
Arbuscular mycorrhizal (AM) fungal extraradical hyphae exude their metabolites into the soil. Root exudate metabolites are affected by plant species and P status. However, the effect of P status on AM hyphal exudate metabolites has been unknown. This study aimed to examine hyphal exudate metabolite composition of two AM fungal species and their response to P deficiency through metabolite profiling. Rhizophagus clarus and R. irregularis were grown in a two-compartment in vitro culture system of Linum usitatissimum roots on solid modified Strullu-Romand medium in combination with two P levels (3 µM (P3) and 30 µM (P30)). Hyphal exudates were collected from the hyphal compartment at 118 days after inoculation (DAI). The metabolite composition of the hyphal exudates was determined by capillary electrophoresis/time-of-flight mass spectrometry, resulting in the identification of a total of 141 metabolites at 118 DAI. In the hyphal exudates of R. clarus, the concentrations of 18 metabolites, including sugars, amino acids, and organic acids, were significantly higher (p < 0.05) under P3 than under P30 conditions. In contrast, the concentrations of 10 metabolites, including sugar and amino acids, in the hyphal exudates of R. irregularis were significantly lower (p < 0.05) under P3 than under P30 conditions. These findings suggest that the extraradical hyphae of AM fungi exude diverse metabolites of which concentrations are affected by P conditions and differ between AM fungal species.
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Data availability
Raw data of this study are available from the corresponding author Keitaro Tawaraya on request.
References
Ahonen-Jonnarth U, van Hees PAW, Lundstrom US, Finlay RD (2000) Organic acids produced by mycorrhizal Pinus sylvestris exposed to elevated aluminium and heavy metal concentration. New Phytol 146:557–567. https://doi.org/10.1046/j.1469-8137.2000.00653.x
Allen MF, Moore TS Jr, Christensen M (1980) Phytohormone changes in Bouteloua gracilis infected by vesicular-arbuscular mycorrhizae: I. Cytokinin increases in the host plant. Can J Bot 58:371–374. https://doi.org/10.1139/b80-038
Bharadwaj DP, Alström S, Lundquist P (2012) Interactions among Glomus irregulare, arbuscular mycorrhizal spore-associated bacteria, and plant pathogens under in vitro conditions. Mycorrhiza 22:437–447. https://doi.org/10.1007/s00572-011-0418-7
Cairney JWG, Burke RM (1998) Extracellular enzyme activities of the ericoid mycorrhizal endophyte Hymenoscyphusericae (Read) Korf & Kernan: their likely roles in decomposition of dead plant tissue in soil. Plant Soil 205:181–192. https://doi.org/10.1023/A:1004376731209
Carvalhais LC, Dennis PG, Fedoseyenko D, Hajirezaei MR, Borris R, Wiren NV (2011) Root exudation of sugar, 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
Chen YP, Rekha PD, Arun AB, Shen FT, Lai WA, Young CC (2006) Phosphate solubilizing bacteria from subtropical soil and their tricalcium phosphate solubilizing abilities. Appl Soil Ecol 32:33–41. https://doi.org/10.1016/j.apsoil.2005.12.002
Declerck S, Strullu DG, Plenchette C (1998) Monoxenic culture of the intraradical forms of Glomus sp. isolated from a tropical ecosystem: a proposed methodology for germplasm collection. Mycologia 90:579–585. https://doi.org/10.1080/00275514.1998.12026946
Della Monica IF, Godeas AM, Scervino JM (2020) In vivo modulation of arbuscular mycorrhizal symbiosis and soil quality by fungal P solubilizers. Microb Ecol 79:21–29. https://doi.org/10.1007/s00248-019-01396-6
Doner LW, Becard G (1991) Solubilization of gellan gels by chelation of cations. Biotechnol Tech 5:25–28. https://doi.org/10.1007/BF00152749
Edayilam N, Montgomery D, Ferguson B, Maroli AS, Martinez N, Powell BA, Tharayil N (2018) Phosphorus stress-induced changes in plant root exudation could potentially facilitate uranium mobilization from stable mineral forms. Environ Sci Technol 52:7652–7662. https://doi.org/10.1021/acs.est.7b05836
Filion M, ST-Arnaud M, Fortin JA, (1999) Direct interaction between the arbuscular mycorrhizal fungus Glomus intraradices and different rhizosphere microorganisms. New Phytol 141:525–533. https://doi.org/10.1046/j.1469-8137.1999.00366.x
Hodge A, Alexander IJ, Gooday GW (1995) Chitinolytic enzymes of pathogenic and ectomycorrhizal fungi. Mycol Res 99:935–941. https://doi.org/10.1016/S0953-7562(09)80752-1
Hooker JE, Piatti P, Cheshire MV, Watson CA (2007) Polysaccharides and monosaccharides in the hyphosphere of the arbuscular mycorrhizal fungi Glomus E3 and Glomus tenue. Soil BiolBiochem 39:680–683. https://doi.org/10.3389/fmicb.2016.00939
Islam MK, Sano A, Majumder MSI, Sakagami JI, Gima S, Hossain MA (2019) Evaluation of organic acid production potential of phosphate solubilizing fungi isolated from soils in Okinawa, Japan. Appl Ecol Environ Res 17: 15191–15201.https://doi.org/10.15666/aeer/1706_1519115201
Jakobsen I, Abbott LK, Robson AD (1992) External hyphae of vesicular-arbuscular mycorrhizal fungi associated with Trifolium subterraneum L. New Phytol 120:371–380. https://doi.org/10.1111/j.1469-8137.1992.tb01077.x
Johansen A, Finlay RD, Olsson PA (1996) Nitrogen metabolism of external hyphae of the arbuscular mycorrhizal fungus Glomus intraradices. New Phytol 133:705–712. https://doi.org/10.1111/j.1469-8137.1996.tb01939.x
Johansson EM, Fransson PMA, Finlay RD, van Hees PAW (2008) Quantitative analysis of root and ectomycorrhizal exudates as a response to Pb, Cd and As stress. Plant Soil 313:39–54. https://doi.org/10.1007/s11104-008-9678-1
Koide RT, Kabir Z (2000) Extraradical hyphae of the mycorrhizal fungus Glomus intraradices can hydrolyse organic phosphate. New Phytol 148:511–517. https://doi.org/10.1046/j.1469-8137.2000.00776.x
Li X, Sun X, Wang G, Amombo E, Zhou X, Du Z, Zhang Y, Xie Y, Fu J (2019) Inoculation with Aspergillus aculeatus alters the performance of perennial ryegrass under phosphorus deficiency. J Am Soc Hortic Sci 144: 182–192. https://doi.org/10.21273/JASHS04581-1s8
Li XL, George E, Marschner H (1991) Extension of the phosphorus depletion zone in VA-mycorrhizal white clover in a calcareous soil. Plant Soil 136:41–48. https://doi.org/10.1007/BF02465218
Martin-Rodriguez JA, Leon-Morcillo R, Vierheilig H, Ocampo JA, Ludwig-Muller J, Garcia-Garrido JM (2011) Ethylene-dependent/ethylene-independent ABA regulation of tomato plants colonized by arbuscular mycorrhiza fungi. New Phytol 190:193–205. https://doi.org/10.1111/j.1469-8137.2010.03610.x
Meier IC, Pritchard SG, Brzostek ER, McCormack ML, Phillips RP (2015) The rhizosphere and hyphosphere differ in their impacts on carbon and nitrogen cycling in forests exposed to elevated CO2. New Phytol 205:1164–1174. https://doi.org/10.1111/nph.13122
Miller RM, Jastrow JD, Reinhardt DR (1995) External hyphal production of vesicular-arbuscular mycorrhizal fungi in pasture and tallgrass prairie communities. Oecologia 103:17–23. https://doi.org/10.1007/BF00328420
Miller SB, Heuberger AL, Broeckling CD, Jahn CE (2019) Non-targeted metabolomics reveals sorghum rhizosphere-associated exudates are influenced by the below ground interaction of substrate and sorghum genotype. Int J Mol Sci 20:431. https://doi.org/10.3390/ijms20020431
Oldroyd GED (2013) Speak, friend and enter: signaling systems that promote beneficial symbiotic associations in plants. Nat Rev Microbiol 11(4):252–263. https://doi.org/10.1038/nrmicro2990
Rodriguez H, Gonzalez T, Goire I, Bashan Y (2004) Gluconic acid production and phosphate solubilization by the plant growth-promoting bacterium Azospirillums pp. Naturwissenschaften 91:552–555. https://doi.org/10.1007/s00114-004-0566-0
Sato T, Hachiya S, Inamura N, Ezawa T, Cheng W, Tawaraya K (2019) Secretion of acid phosphatase from extraradical hyphae of the arbuscular mycorrhizal fungus Rhizophagus clarus is regulated in response to phosphate availability. Mycorrhiza 29:599–605. https://doi.org/10.1007/s00572-019-00923-0
Sato T, Ezawa T, Cheng W, Tawaraya K (2015) Release of acid phosphatase from extraradical hyphae of arbuscular mycorrhizal fungus Rhizophagus clarus. J Soil Sci Plant Nutr 61:269–274. https://doi.org/10.1080/00380768.2014.993298
St-Arnaud M, Hamel C, Vimard B, Caron M, Fortin JA (1996) Enhanced hyphal growth and spore production of the arbuscular mycorrhizal fungus Glomus intraradices in an in vitro system in the absence of host roots. Mycol Res 100:328–332. https://doi.org/10.1016/S0953-7562(96)80164-X
Sumner LW, Mendes P, Dixon RA (2003) Plant metabolomics: large-scale phytochemistry in the functional genomics era. Phytochemistry 62(6):817–836. https://doi.org/10.1016/s0031-9422(02)00708-2
Sung J, Lee S, Lee Y, Ha S, Song B, Kim T, Waters BM, Krishnan HB (2015) Metabolic profiling from leaves and roots of tomato (Solanum lycopersicum L.) plants grown under nitrogen, phosphorus or potassium deficient condition. Plant Sci 241:55–64. https://doi.org/10.1016/j.plantsci.2015.09.027
Tawaraya K, Horie R, Wagatsuma T, Saito K, Oikawa A (2018) Metabolite profiling of shoot extract, root extract, and root exudate of rice under nitrogen and phosphorus deficiency. J Soil Sci Plant Nutr 64:312–322. https://doi.org/10.1080/00380768.2018.1476828
Tawaraya K, Horie R, Saito K, Wagatsuma T, Saito K, Oikawa A (2014) Metabolite profiling of root exudates of common bean under phosphorus deficiency. Metabolites 4:599–611. https://doi.org/10.3390/metabo4030599
Tawaraya K, Horie R, Shinano T, Wagatsuma T, Saito K, Oikawa A (2014) Metabolite profiling of soybean root exudates under phosphorus deficiency. J Soil Sci Plant Nutr 60:679–694. https://doi.org/10.1080/00380768.2014.945390
Tawaraya K, Horie R, Saito A, Shinano T, Wagatsuma T, Saito K, Oikawa A (2013) Metabolite profiling of shoot extracts, root extracts, and root exudates of rice plant under phosphorus deficiency. J Plant Nutr 36:1138–1159. https://doi.org/10.1080/01904167.2013.780613
Tawaraya K, Naito M, Wagatsuma T (2006) Solubilization of insoluble inorganic phosphate by hyphal exudates of arbuscular mycorrhizal fungi. J Plant Nutr 29:657–665. https://doi.org/10.1080/01904160600564428
Toljander JF, Artursson V, Paul LR, Jansson JK, Finlay RD (2006) Attachment of different soil bacteria to arbuscular mycorrhizal fungal extraradical hyphae is determined by hyphal vitality and fungal species. FEMS Microbiol Lett 254:34–40. https://doi.org/10.1111/j.1574-6968.2005.00003.x
Toljander JF, Lindahl BD, Paul LR, Elfstrand M, Finlay RD (2007) Influence of arbuscular mycorrhizal mycelial exudates on soil bacterial growth and community structure. FEMS Microbiol Ecol 61:295–304. https://doi.org/10.1111/j.1574-6941.2007.00337.x
Toro M, Azcón R, Barea J (1997) Improvement of arbuscular mycorrhizal development by inoculation of soil with phosphate-solubilizing rhizobacteria to improve rock phosphate bioavaiability (32P) and nutrient cycling. Appl Eniviron Microbiol 63:4408–4412. https://doi.org/10.1128/AEM.63.11.4408-4412.1997
Van Hees PAW, Rosling A, Essen S, Godbold DL, Jones DL, Finlay RD (2006) Oxalate and ferricrocin exudation by the extramatrical mycelium of an ectomycorrhizal fungus in symbiosis with Pinus sylvestris. New Phytol 169:367–378. https://doi.org/10.1111/j.1469-8137.2005.01600.x
Villegas J, Fortin JA (2001) Phosphorus solubilization and pH changes as a result of the interactions between soil bacteria and arbuscular mycorrhizal fungi on a medium containing NH4+ as nitrogen source. Can J Bot 79:865–870. https://doi.org/10.1139/b01-069
Witzel K, Strehmel N, Baldermann S, Neugart S, Becker Y, Becker M, Berger B, Scheel D, Grosch R, Schreiner M, Ruppel S (2017) Arabidopsis thaliana root and root exudate metabolism is altered by the growth-promoting bacterium Kosakonia radicincitans DSM 16656T. Plant Soil 419:557–573. https://doi.org/10.1007/s11104-017-3371-1
Wu M, Wei Q, Xu L, Li H, Oelmüller R, Zhang W (2018) Piriformospora indica enhances phosphorus absorption by stimulating acid phosphatase activities and organic acid accumulation in Brassica napus. Plant Soil 432:333–344. https://doi.org/10.1007/s11104-018-3795-2
Yang C, Zhao W, Wang Y, Zhang L, Huang S, Lin J (2020) Metabolomics analysis reveals the alkali tolerance mechanis in Puccinellia tenuiflora plants inoculated with arbuscular mycorrhizal fungi. Microorganism 8:327. https://doi.org/10.3390/microorganisms8030327
Zhang L, Peng Y, Zhou J, George TS, Feng Gu (2020) Addition of fructose to the maize hyphosphere increases phosphatase activity by changing bacterial community structure. Soil BiolBiochem 142(107724):1–9. https://doi.org/10.1016/j.soilbio.2020.107724
Zhang L, Fan J, Feng G, Declerck S (2018) The arbuscularmycorrhizal fungus Rhizophagus irregularis MUCL 43194 induces the gene expression of citrate synthase in the tricarboxylic acid cycle of the phosphate-solubilizing bacterium Rahnella aquatilis HX2. Mycorrhiza 29:69–75. https://doi.org/10.1007/s00572-018-0871-7
Zhang L, Xu M, Liu Y, Zhang F, Hodge A, Feng G (2016) Carbon and phosphorus exchange may enable cooperation between an arbuscular mycorrhizal fungus and a phospahate-solybilizing bacterium. New Phytol 210:1022–1032. https://doi.org/10.1111/nph.13838
Zhou Y, Neuhäuser B, Neumann G, Ludewig U (2020) LaALMT1 mediates malate release from phosphorus-deficient white lupin root tips and metal root to shoot translocation. Plant Cell Environ 43:1691–1706. https://doi.org/10.1111/pce.13762
Ziegler J, Schmidt S, Chutia R, Muller J, Bottcher C, Strehmel N, Scheel D, Abel S (2016) Non-targeted profiling of semi-polar metabolites in Arabidopsis root exudates uncovers a role for coumarin secretion and lignification during the local response to phosphate limitation. J Exp Bot 67:1421–1432. https://doi.org/10.1093/jxb/erv539
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This study was supported by JSPS KAKENHI Grant Numbers JP15K07332 and JP18K05368.
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NL, NI, Tantriani, TS and KT designed the study and performed the experiments. NL, NI, Tantriani, TS AO, WC, and KT analyzed the data. NL and KT wrote the manuscript. All authors read and approved the final manuscript.
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Luthfiana, N., Inamura, N., Tantriani et al. Metabolite profiling of the hyphal exudates of Rhizophagus clarus and Rhizophagus irregularis under phosphorus deficiency. Mycorrhiza 31, 403–412 (2021). https://doi.org/10.1007/s00572-020-01016-z
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DOI: https://doi.org/10.1007/s00572-020-01016-z