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Fungal lipochitooligosaccharide symbiotic signals in arbuscular mycorrhiza

Abstract

Arbuscular mycorrhiza (AM) is a root endosymbiosis between plants and glomeromycete fungi. It is the most widespread terrestrial plant symbiosis, improving plant uptake of water and mineral nutrients. Yet, despite its crucial role in land ecosystems, molecular mechanisms leading to its formation are just beginning to be unravelled. Recent evidence suggests that AM fungi produce diffusible symbiotic signals. Here we show that Glomus intraradices secretes symbiotic signals that are a mixture of sulphated and non-sulphated simple lipochitooligosaccharides (LCOs), which stimulate formation of AM in plant species of diverse families (Fabaceae, Asteraceae and Umbelliferae). In the legume Medicago truncatula these signals stimulate root growth and branching by the symbiotic DMI signalling pathway. These findings provide a better understanding of the evolution of signalling mechanisms involved in plant root endosymbioses and will greatly facilitate their molecular dissection. They also open the way to using these natural and very active molecules in agriculture.

Figure 1: Detection and characterization of LCOs in mycorrhized carrot root exudates.
Figure 2: Detection and characterization of LCOs in germinating G. intraradices spore exudates.
Figure 3: Chemical structures of natural and synthetic Myc-LCOs.
Figure 4: Effect of Myc-LCOs on mycorrhization by G. intraradices.
Figure 5: Synthetic Myc-LCOs stimulate M. truncatula root branching by the DMI pathway.

References

  1. 1

    Smith, S. E. & Read, D. J. in Mycorrhizal Symbiosis. 145–187 (Academic Press, 2008)

    Google Scholar 

  2. 2

    Harrison, M. J. Signaling in the arbuscular mycorrhizal symbiosis. Annu. Rev. Microbiol. 59, 19–42 (2005)

    CAS  Article  Google Scholar 

  3. 3

    Oldroyd, G. E. & Downie, J. A. Coordinating nodule morphogenesis with rhizobial infection in legumes. Annu. Rev. Plant Biol. 59, 519–546 (2008)

    CAS  Article  Google Scholar 

  4. 4

    Stacey, G., Libault, M., Brechenmacher, L., Wan, J. & May, G. D. Genetics and functional genomics of legume nodulation. Curr. Opin. Plant Biol. 9, 110–121 (2006)

    CAS  Article  Google Scholar 

  5. 5

    Masson-Boivin, C., Giraud, E., Perret, X. & Batut, J. Establishing nitrogen-fixing symbiosis with legumes: how many rhizobium recipes? Trends Microbiol. 17, 458–466 (2009)

    CAS  Article  Google Scholar 

  6. 6

    Dénarié, J., Debellé, F. & Promé, J. C. Rhizobium lipo-chitooligosaccharide nodulation factors: signaling molecules mediating recognition and morphogenesis. Annu. Rev. Biochem. 65, 503–535 (1996)

    Article  Google Scholar 

  7. 7

    Parniske, M. Arbuscular mycorrhiza: the mother of plant root endosymbioses. Nature Rev. Microbiol. 6, 763–775 (2008)

    CAS  Article  Google Scholar 

  8. 8

    Oldroyd, G. E., Harrison, M. J. & Paszkowski, U. Reprogramming plant cells for endosymbiosis. Science 324, 753–754 (2009)

    ADS  CAS  Article  Google Scholar 

  9. 9

    Catoira, R. et al. Four genes of Medicago truncatula controlling components of a nod factor transduction pathway. Plant Cell 12, 1647–1666 (2000)

    CAS  Article  Google Scholar 

  10. 10

    Wais, R. J. et al. Genetic analysis of calcium spiking responses in nodulation mutants of Medicago truncatula . Proc. Natl Acad. Sci. USA 97, 13407–13412 (2000)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Kosuta, S. et al. Differential and chaotic calcium signatures in the symbiosis signaling pathway of legumes. Proc. Natl Acad. Sci. USA 105, 9823–9828 (2008)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Oldroyd, G. E. & Downie, J. A. Nuclear calcium changes at the core of symbiosis signalling. Curr. Opin. Plant Biol. 9, 351–357 (2006)

    CAS  Article  Google Scholar 

  13. 13

    Riely, B. K., Ané, J. M., Penmetsa, R. V. & Cook, D. R. Genetic and genomic analysis in model legumes bring Nod-factor signaling to center stage. Curr. Opin. Plant Biol. 7, 408–413 (2004)

    CAS  Article  Google Scholar 

  14. 14

    Weidmann, S. et al. Fungal elicitation of signal transduction-related plant genes precedes mycorrhiza establishment and requires the dmi3 gene in Medicago truncatula . Mol. Plant Microbe Interact. 17, 1385–1393 (2004)

    CAS  Article  Google Scholar 

  15. 15

    Oláh, B., Brière, C., Bécard, G., Dénarié, J. & Gough, C. Nod factors and a diffusible factor from arbuscular mycorrhizal fungi stimulate lateral root formation in Medicago truncatula via the DMI1/DMI2 signalling pathway. Plant J. 44, 195–207 (2005)

    Article  Google Scholar 

  16. 16

    Gutjahr, C. et al. Presymbiotic factors released by the arbuscular mycorrhizal fungus Gigaspora margarita induce starch accumulation in Lotus japonicus roots. New Phytol. 183, 53–61 (2009)

    CAS  Article  Google Scholar 

  17. 17

    Kuhn, H., Kuster, H. & Requena, N. Membrane steroid-binding protein 1 induced by a diffusible fungal signal is critical for mycorrhization in Medicago truncatula . New Phytol. 185, 716–733 (2010)

    CAS  Article  Google Scholar 

  18. 18

    Remy, W., Taylor, T. N., Hass, H. & Kerp, H. Four hundred-million-year-old vesicular arbuscular mycorrhizae. Proc. Natl Acad. Sci. USA 91, 11841–11843 (1994)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Sprent, J. I. Evolving ideas of legume evolution and diversity: a taxonomic perspective on the occurrence of nodulation. New Phytol. 174, 11–25 (2007)

    CAS  Article  Google Scholar 

  20. 20

    Price, N. P. et al. Broad-host-range Rhizobium species strain NGR234 secretes a family of carbamoylated, and fucosylated, nodulation signals that are O-acetylated or sulphated. Mol. Microbiol. 6, 3575–3584 (1992)

    CAS  Article  Google Scholar 

  21. 21

    Andriankaja, A. et al. AP2-ERF transcription factors mediate Nod factor-dependent Mt ENOD11 activation in root hairs via a novel cis-regulatory motif. Plant Cell 19, 2866–2885 (2007)

    CAS  Article  Google Scholar 

  22. 22

    D’Haeze, W. & Holsters, M. Nod factor structures, responses, and perception during initiation of nodule development. Glycobiology 12, 79R–105R (2002)

    Article  Google Scholar 

  23. 23

    Roche, P., Lerouge, P., Ponthus, C. & Promé, J. C. Structural determination of bacterial nodulation factors involved in the Rhizobium meliloti-alfalfa symbiosis. J. Biol. Chem. 266, 10933–10940 (1991)

    CAS  PubMed  Google Scholar 

  24. 24

    Ardourel, M. et al. Rhizobium meliloti lipooligosaccharide nodulation factors: different structural requirements for bacterial entry into target root hair cells and induction of plant symbiotic developmental responses. Plant Cell 6, 1357–1374 (1994)

    CAS  Article  Google Scholar 

  25. 25

    Spaink, H. P. et al. Structural identification of metabolites produced by the NodB and NodC proteins of Rhizobium leguminosarum . Mol. Microbiol. 13, 821–831 (1994)

    CAS  Article  Google Scholar 

  26. 26

    Spaink, H. P., Wijfjes, A. H. & Lugtenberg, B. J. Rhizobium NodI and NodJ proteins play a role in the efficiency of secretion of lipochitin oligosaccharides. J. Bacteriol. 177, 6276–6281 (1995)

    CAS  Article  Google Scholar 

  27. 27

    Samain, E., Drouillard, S., Heyraud, A., Driguez, H. & Geremia, R. A. Gram-scale synthesis of recombinant chitooligosaccharides in Escherichia coli . Carbohydr. Res. 302, 35–42 (1997)

    CAS  Article  Google Scholar 

  28. 28

    Samain, E., Chazalet, V. & Geremia, R. A. Production of O-acetylated and sulfated chitooligosaccharides by recombinant Escherichia coli strains harboring different combinations of nod genes. J. Biotechnol. 72, 33–47 (1999)

    CAS  Article  Google Scholar 

  29. 29

    Ohsten Rasmussen, M., Hogg, B., Bono, J. J., Samain, E. & Driguez, H. New access to lipo-chitooligosaccharide nodulation factors. Org. Biomol. Chem. 2, 1908–1910 (2004)

    Article  Google Scholar 

  30. 30

    Liu, J. et al. Transcript profiling coupled with spatial expression analyses reveals genes involved in distinct developmental stages of an arbuscular mycorrhizal symbiosis. Plant Cell 15, 2106–2123 (2003)

    CAS  Article  Google Scholar 

  31. 31

    Bécard, G. & Fortin, J. A. Early events of vesicular–arbuscular mycorrhiza formation in Ri T-DNA transformed roots. New Phytol. 108, 211–218 (1988)

    Article  Google Scholar 

  32. 32

    Arrighi, J. F. et al. The Medicago truncatula lysin motif-receptor-like kinase gene family includes NFP and new nodule-expressed genes. Plant Physiol. 142, 265–279 (2006)

    CAS  Article  Google Scholar 

  33. 33

    Smit, P. et al. Medicago LYK3, an entry receptor in rhizobial nodulation factor signaling. Plant Physiol. 145, 183–191 (2007)

    CAS  Article  Google Scholar 

  34. 34

    Ané, J. M. et al. Medicago truncatula DMI1 required for bacterial and fungal symbioses in legumes. Science 303, 1364–1367 (2004)

    ADS  Article  Google Scholar 

  35. 35

    Endre, G. et al. A receptor kinase gene regulating symbiotic nodule development. Nature 417, 962–966 (2002)

    ADS  CAS  Article  Google Scholar 

  36. 36

    Lévy, J. et al. A putative Ca2+ and calmodulin-dependent protein kinase required for bacterial and fungal symbioses. Science 303, 1361–1364 (2004)

    ADS  Article  Google Scholar 

  37. 37

    Smit, P. et al. NSP1 of the GRAS protein family is essential for rhizobial Nod factor-induced transcription. Science 308, 1789–1791 (2005)

    ADS  CAS  Article  Google Scholar 

  38. 38

    Kalo, P. et al. Nodulation signaling in legumes requires NSP2, a member of the GRAS family of transcriptional regulators. Science 308, 1786–1789 (2005)

    ADS  CAS  Article  Google Scholar 

  39. 39

    Kosuta, S. et al. A diffusible factor from arbuscular mycorrhizal fungi induces symbiosis-specific MtENOD11 expression in roots of Medicago truncatula . Plant Physiol. 131, 952–962 (2003)

    CAS  Article  Google Scholar 

  40. 40

    Gutjahr, C., Casieri, L. & Paszkowski, U. Glomus intraradices induces changes in root system architecture of rice independently of common symbiosis signaling. New Phytol. 182, 829–837 (2009)

    Article  Google Scholar 

  41. 41

    Chen, C., Gao, M., Liu, J. & Zhu, H. Fungal symbiosis in rice requires an ortholog of a legume common symbiosis gene encoding a Ca2+/calmodulin-dependent protein kinase. Plant Physiol. 145, 1619–1628 (2007)

    CAS  Article  Google Scholar 

  42. 42

    Chen, C., Ané, J. M. & Zhu, H. OsIPD3, an ortholog of the Medicago truncatula DMI3 interacting protein IPD3, is required for mycorrhizal symbiosis in rice. New Phytol. 180, 311–315 (2008)

    CAS  Article  Google Scholar 

  43. 43

    Wang, B. et al. Presence of three mycorrhizal genes in the common ancestor of land plants suggests a key role of mycorrhizas in the colonization of land by plants. New Phytol. 186, 514–525 (2010)

    Article  Google Scholar 

  44. 44

    Chabot, S., Bécard, G. & Piche, Y. The life cycle of Glomus intraradices in root organ culture. Mycologia 84, 315–321 (1992)

    Article  Google Scholar 

  45. 45

    Ben Amor, B. et al. The NFP locus of Medicago truncatula controls an early step of Nod factor signal transduction upstream of a rapid calcium flux and root hair deformation. Plant J. 34, 495–506 (2003)

    CAS  Article  Google Scholar 

  46. 46

    Giovannetti, M. & Mosse, B. An evaluation of techniques for measuring vesicular-arbuscular mycorrhizal infection in roots. New Phytol. 84, 489–500 (1980)

    Article  Google Scholar 

  47. 47

    Hirsch, S. et al. GRAS proteins form a DNA binding complex to induce gene expression during nodulation signaling in Medicago truncatula . Plant Cell 21, 545–557 (2009)

    CAS  Article  Google Scholar 

  48. 48

    Journet, E. P. et al. Medicago truncatula ENOD11: a novel RPRP-encoding early nodulin gene expressed during mycorrhization in arbuscule-containing cells. Mol. Plant Microbe Interact. 14, 737–748 (2001)

    CAS  Article  Google Scholar 

  49. 49

    Poinsot, V. et al. Unusual methyl-branched α,β-unsaturated acyl chain substitutions in the Nod factors of an arctic rhizobium, Mesorhizobium sp. strain N33 (Oxytropis arctobia). J. Bacteriol. 183, 3721–3728 (2001)

    CAS  Article  Google Scholar 

  50. 50

    Gomez-Roldan, V. et al. Strigolactone inhibition of shoot branching. Nature 455, 189–194 (2008)

    ADS  CAS  Article  Google Scholar 

  51. 51

    Demont, N., Debellé, F., Aurelle, H., Dénarié, J. & Promé, J. C. Role of the Rhizobium meliloti nodF and nodE genes in the biosynthesis of lipo-oligosaccharidic nodulation factors. J. Biol. Chem. 268, 20134–20142 (1993)

    CAS  PubMed  Google Scholar 

  52. 52

    Olsthoorn, M. M. et al. Novel branched nod factor structure results from alpha-(1→3) fucosyl transferase activity: the major lipo-chitin oligosaccharides from Mesorhizobium loti strain NZP2213 bear an alpha-(1→3) fucosyl substituent on a nonterminal backbone residue. Biochemistry 37, 9024–9032 (1998)

    CAS  Article  Google Scholar 

  53. 53

    Hewitt, E. J. Sand and Water Culture Methods Used in the Study of Plant Nutrition (Commonwealth Agricultural Bureaux, 1966)

    Google Scholar 

  54. 54

    Vierheilig, H., Coughlan, A. P., Wyss, U. & Piche, Y. Ink and vinegar, a simple staining technique for arbuscular-mycorrhizal fungi. Appl. Environ. Microbiol. 64, 5004–5007 (1998)

    CAS  PubMed  PubMed Central  Google Scholar 

  55. 55

    R Development Core Team R: A Language and Environment for Statistical Computing. (R Foundation for Statistical Computing, 2009)

    Google Scholar 

Download references

Acknowledgements

This work was supported in part by a grant from EMD Crop BioScience and by the Charles-Léopold Mayer Prize (2005) attributed to J.D. by the French Academy of Sciences. M.G. was supported by an Institut National de la Recherche Agronomique fellowship. We are grateful to C. Gough and G. Oldroyd for providing seeds of M. truncatula mutants, and to S. Roy and J. Loubradou for providing sterile mycorrhized roots and spores of G. intraradices. The UPLC/QToF mass spectrometer was made available to us by the Institut des Technologies Avancées du Vivant and the QTRAP mass spectrometer by the Metabolomics and fluxomics platform (MetaToul). We thank S. Danoun for her help with UPLC/QToF mass spectrometry, F. Letisse for his advice and help in setting up experiments on the QTrap mass spectrometer, P. Lavedan for NMR measurements, C. Hervé for her advice and help for quantitative PCR with reverse transcription experiments and C. Brière for his advice for statistical analysis. We thank C. Gough, J. Cullimore and G. Oldroyd for their comments on this manuscript.

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F.M. coordinated experiments for all bioassays, designed and performed the VsHab assay, and designed and performed M. truncatula bioassays with O.A. O.A. and V.P. purified fungal Myc-LCOs. M.G. and A.H. prepared germinating spores. L.C. and A.H. performed the ENOD11 bioassay. A.H. designed and performed mycorrhization tests on carrots and Tagetes. O.A. performed statistical analyses. F.M. and D.G. extracted Myc-LCO analogues from rhizobial mutant cultures, and E.A.M. and H.D. synthesized Myc-LCOs by the cell factory technique. For mass spectrometry, UPLC/QToF was performed by V.P. and V.P.-P., and QTRAP by V.P.-P. Quantitative PCR with reverse transcription experiments were designed and analysed by A.N. and realized by D.F. and O.A. G.B. supervised devising, planning and interpreting experiments with AM fungal material (material production, spore germination and mycorrhization tests). J.D. conceived and directed the project, and wrote the manuscript with the help of F.M., V.P. and G.B.

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Correspondence to Jean Dénarié.

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Maillet, F., Poinsot, V., André, O. et al. Fungal lipochitooligosaccharide symbiotic signals in arbuscular mycorrhiza. Nature 469, 58–63 (2011). https://doi.org/10.1038/nature09622

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