Abstract
Arbuscular mycorrhizal fungi (AMF) are symbiotic soil fungi that are intimately associated with the roots of the majority of land plants. They colonise the interior of the roots and the hyphae extend into the soil. It is well known that bacterial colonisation of the rhizosphere can be crucial for many pathogenic as well as symbiotic plant–microbe interactions. However, although bacteria colonising the extraradical AMF hyphae (the hyphosphere) might be equally important for AMF symbiosis, little is known regarding which bacterial species would colonise AMF hyphae. In this study, we investigated which bacterial communities might be associated with AMF hyphae. As bacterial-hyphal attachment is extremely difficult to study in situ, we designed a system to grow AMF hyphae of Glomus intraradices and Glomus proliferum and studied which bacteria separated from an agricultural soil specifically attach to the hyphae. Characterisation of attached and non-attached bacterial communities was performed using terminal restriction fragment length polymorphism and clone library sequencing of 16S ribosomal RNA (rRNA) gene fragments. For all experiments, the composition of hyphal attached bacterial communities was different from the non-attached communities, and was also different from bacterial communities that had attached to glass wool (a non-living substratum). Analysis of amplified 16S rRNA genes indicated that in particular bacteria from the family of Oxalobacteraceae were highly abundant on AMF hyphae, suggesting that they may have developed specific interactions with the fungi.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
References
Andrade G, Linderman RG, Bethlenfalvay GJ . (1998). Bacterial associations with the mycorrhizosphere and hyphosphere of the arbuscular mycorrhizal fungus Glomus mosseae. Plant Soil 202: 79–87.
Artursson V, Jansson JK . (2003). Use of bromodeoxyuridine immunocapture to identify active bacteria associated with arbuscular mycorrhizal hyphae. Appl Environ Microbiol 69: 6208–6215.
Azcon-Aguilar C, Barea JM . (1996). Arbuscular mycorrhizas and biological control of soil-borne plant pathogens—an overview of the mechanisms involved. Mycorrhiza 6: 457–464.
Becard G, Fortin JA . (1988). Early events of vesicular arbuscular mycorrhiza formation on Ri T-DNA transformed roots. New Phytol 108: 211–218.
Bianciotto V, Andreotti S, Balestrini R, Bonfante P, Perotto S . (2001). Mucoid mutants of the biocontrol strain Pseudomonas fluorescens CHA0 show increased ability in biofilm formation on mycorrhizal and nonmycorrhizal carrot roots. Mol Plant Microbe Interact 14: 255–260.
Bianciotto V, Minerdi D, Perotto S, Bonfante P . (1996). Cellular interactions between arbuscular mycorrhizal fungi and rhizosphere bacteria. Protoplasma 193: 123–131.
Bonfante P, Anca IA . (2009). Plants, mycorrhizal fungi, and bacteria: a network of interactions. Annu Rev Microbiol 63: 363–383.
Borowicz VA . (2001). Do arbuscular mycorrhizal fungi alter plant-pathogen relations? Ecology 82: 3057–3068.
Brucker RM, Harris RN, Schwantes CR, Gallaher TN, Flaherty DC, Lam BA et al. (2008). Amphibian chemical defense: antifungal metabolites of the microsymbiont Janthinobacterium lividum on the salamander Plethodon cinereus. J Chem Ecol 34: 1422–1429.
Budi SW, van Tuinen D, Martinotti G, Gianinazzi S . (1999). Isolation from the Sorghum bicolor mycorrhizosphere of a bacterium compatible with arbuscular mycorrhiza development and antagonistic towards soilborne fungal pathogens. Appl Environ Microbiol 65: 5148–5150.
Bürgmann H, Pesaro M, Widmer F, Zeyer J . (2001). A strategy for optimizing quality and quantity of DNA extracted from soil. J Microbiol Methods 45: 7–21.
Cruz AF, Horii S, Ochiai S, Yasuda A, Ishii T . (2008). Isolation and analysis of bacteria associated with spores of Gigaspora margarita. J Appl Microbiol 104: 1711–1717.
Davies FT, Potter JR, Linderman RG . (1993). Drought resistance of mycorrhizal pepper plants independent of leaf P concentration—response in gas exchange and water relations. Physiol Plant 87: 45–53.
de Boer W, Leveau JHJ, Kowalchuk GA, Gunnewiek P, Abeln ECA, Figge MJ et al. (2004). Collimonas fungivorans gen. nov., sp nov., a chitinolytic soil bacterium with the ability to grow on living fungal hyphae. Int J Syst Evol Microbiol 54: 857–864.
Declerck S, Cranenbrouck S, Dalpe Y, Seguin S, Grandmougin-Ferjani A, Fontaine J et al. (2000). Glomus proliferum sp nov.: a description based on morphological, biochemical, molecular and monoxenic cultivation data. Mycologia 92: 1178–1187.
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.
Frey-Klett P, Garbaye J, Tarkka M . (2007). The mycorrhiza helper bacteria revisited. New Phytol 176: 22–36.
Garbaye J . (1994). Helper bacteria: a new dimension to the mycorrhizal symbiosis. New Phytol 128: 197–210.
Gleave AP, Taylor RK, Morris BAM, Greenwood DR . (1995). Cloning and sequencing of a gene encoding the 69-kDa extracellular chitinase of Janthinobacterium lividum. FEMS Microbiol Ecol 131: 279–288.
Green SJ, Michel FC, Hadar Y, Minz D . (2007). Contrasting patterns of seed and root colonization by bacteria from the genus Chryseobacterium and from the family Oxalobacteraceae. ISME J 1: 291–299.
Izumi H, Anderson IC, Alexander IJ, Killham K, Moore ERB . (2006). Endobacteria in some ectomycorrhiza of Scots pine (Pinus sylvestris). FEMS Microbiol Ecol 56: 34–43.
Johansson JF, Paul LR, Finlay RD . (2004). Microbial interactions in the mycorrhizosphere and their significance for sustainable agriculture. FEMS Microbiol Ecol 48: 1–13.
Koch AM, Kuhn G, Fontanillas P, Fumagalli L, Goudet J, Sanders IR . (2004). High genetic variability and low local diversity in a population of arbuscular mycorrhizal fungi. PNAS 101: 2369–2374.
Lane DJ . (1991). 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds). Nucleic Acid Techniques in Bacterial Systematics. John Wiley & Sons: New York pp 115–175.
Lincoln SP, Fermor TR, Tindall BJ . (1999). Janthinobacterium agaricidamnosum sp nov., a soft rot pathogen of Agaricus bisporus. Int J Syst Bacteriol 49: 1577–1589.
Mansfeld-Giese K, Larsen J, Bodker L . (2002). Bacterial populations associated with mycelium of the arbuscular mycorrhizal fungus Glomus intraradices. FEMS Microbiol Ecol 41: 133–140.
Marschner P, Baumann K . (2003). Changes in bacterial community structure induced by mycorrhizal colonisation in split-root maize. Plant Soil 251: 279–289.
Mayo K, Davis RE, Motta J . (1986). Stimulation of germination of spores of Glomus versiforme by spore-associated bacteria. Mycologia 78: 426–431.
Muyzer G, de Waal EC, Uitterlinden AG . (1993). Profiling of complex microbial-populations by denaturing gradient gel-electrophoresis analysis of polymerase chain reaction-amplified genes-coding for 16S ribosomal-RNA. Appl Environ Microbiol 59: 695–700.
Newsham KK, Fitter AH, Watkinson AR . (1995). Multi-functionality and biodiversity in arbuscular mycorrhizas. Trends Ecol Evol 10: 407–411.
Offre P, Pivato B, Mazurier S, Siblot S, Berta G, Lemanceau P et al. (2008). Microdiversity of Burkholderiales associated with mycorrhizal and nonmycorrhizal roots of Medicago truncatula. FEMS Microbiol Ecol 65: 180–192.
Offre P, Pivato B, Siblot S, Gamalero E, Corberand T, Lemanceau P et al. (2007). Identification of bacterial groups preferentially associated with mycorrhizal roots of Medicago truncatula. Appl Environ Microbiol 73: 913–921.
Pivato B, Offre P, Marchelli S, Barbonaglia B, Mougel C, Lemanceau P et al. (2009). Bacterial effects on arbuscular mycorrhizal fungi and mycorrhiza development as influenced by the bacteria, fungi, and host plant. Mycorrhiza 19: 81–90.
Ravnskov S, Nybroe O, Jakobsen I . (1999). Influence of an arbuscular mycorrhizal fungus on Pseudomonas fluorescens DF57 in rhizosphere and hyphosphere soil. New Phytol 142: 113–122.
Requena N, Jimenez I, Toro M, Barea JM . (1997). Interactions between plant-growth-promoting rhizobacteria (PGPR), arbuscular mycorrhizal fungi and Rhizobium spp. in the rhizosphere of Anthyllis cytisoides, a model legume for revegetation in mediterranean semi-arid ecosystems. New Phytol 136: 667–677.
Rillig MC, Mummey DL, Ramsey PW, Klironomos JN, Gannon JE . (2006). Phylogeny of arbuscular mycorrhizal fungi predicts community composition of symbiosis-associated bacteria. FEMS Microbiol Ecol 57: 389–395.
Smith SE, Read DJ . (1997). Mycorrhizal Symbiosis, 2nd edn. Academic Press: London.
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.
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 Ecol 254: 34–40.
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.
Trappe JM . (1987). Phylogenetic and ecological aspects of mycotrophy in the angiosperms from an evolutionary standpoint. In: Safir GR (eds). Ecophysiology of VA Mycorrhizal Plants. CRC Press: Boca Raton, pp 5–25.
Tylka GL, Hussey RS, Roncadori RW . (1991). Axenic germination of vesicular-arbuscular mycorrhizal fungi – effects of selected Streptomyces-species. Phytopathology 81: 754–759.
Wamberg C, Christensen S, Jakobsen I, Muller AK, Sørensen SJ . (2003). The mycorrhizal fungus (Glomus intraradices) affects microbial activity in the rhizosphere of pea plants (Pisum sativum). Soil Biol Biochem 35: 1349–1357.
Wang Q, Garrity GM, Tiedje JM, Cole JR . (2007). Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73: 5261–5267.
Weller DM . (2007). Pseudomonas biocontrol agents of soilborne pathogens: looking back over 30 years. Phytopathology 97: 250–256.
Whipps JM . (2001). Microbial interactions and biocontrol in the rhizosphere. J Exp Bot 52: 487–511.
Wyss P, Boller T, Wiemken A . (1992). Testing the effect of biological control agents on the formation of vesicular arbuscular mycorrhiza. Plant Soil 147: 159–162.
Xavier LJC, Germida JJ . (2002). Response of lentil under controlled conditions to co-inoculation with arbuscular mycorrhizal fungi and rhizobia varying in efficacy. Soil Biol Biochem 34: 181–188.
Xavier LJC, Germida JJ . (2003). Bacteria associated with Glomus clarum spores influence mycorrhizal activity. Soil Biol Biochem 35: 471–478.
Acknowledgements
We thank Hans-Rudolf Pfeiffer from the Faculty of Geosciences and the Environment at the University of Lausanne for chemical analysis of the soil that was used to extract bacteria. We also thank the reviewers for their useful comments on the paper. This research was supported by a FBM Interdisciplinary grant from the University of Lausanne.
Author information
Authors and Affiliations
Corresponding author
Additional information
Supplementary Information accompanies the paper on The ISME Journal website
Supplementary information
Rights and permissions
About this article
Cite this article
Scheublin, T., Sanders, I., Keel, C. et al. Characterisation of microbial communities colonising the hyphal surfaces of arbuscular mycorrhizal fungi. ISME J 4, 752–763 (2010). https://doi.org/10.1038/ismej.2010.5
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/ismej.2010.5
Keywords
This article is cited by
-
Core hyphosphere microbiota of Fusarium oxysporum f. sp. niveum
Environmental Microbiome (2024)
-
A symbiotic footprint in the plant root microbiome
Environmental Microbiome (2023)
-
Short-Term Evaluation of the Spatial Distribution of Trophic Groups of Amoebae in the Rhizosphere of Zea mays Inoculated with Rhizophagus intraradices
Microbial Ecology (2023)
-
Hyphosphere microbiome of arbuscular mycorrhizal fungi: a realm of unknowns
Biology and Fertility of Soils (2023)
-
Disentangling arbuscular mycorrhizal fungi and bacteria at the soil-root interface
Mycorrhiza (2023)
