Advertisement

Journal of Plant Research

, Volume 132, Issue 5, pp 641–653 | Cite as

ERN1 and CYCLOPS coordinately activate NIN signaling to promote infection thread formation in Lotus japonicus

  • Meng Liu
  • Takashi Soyano
  • Koji Yano
  • Makoto Hayashi
  • Masayoshi KawaguchiEmail author
Regular Paper
  • 763 Downloads

Abstract

Legumes engage in symbiosis with nitrogen-fixing soil bacteria, collectively called rhizobia, under nitrogen-limited conditions. In many legumes, the root invasion of rhizobia is mediated by infection threads (ITs), tubular invaginations of the host cell wall and plasma membrane, developed from infection foci of deformed root hairs. IT formation is regulated by a series of signal transduction in host root. Nodulation signals activate the host transcription factor (TF), CYCLOPS, which directly induces expression of two TF genes, ERF REQUIRED FOR NODULATION1 (ERN1) and NODULE INCEPTION (NIN), essential for IT development. Here, we explored the relationship among these three symbiotic TF genes in the model legume Lotus japonicus and examined how their interplay contributes to IT formation. qRT-PCR analysis showed that NIN expression induced by rhizobial infection was attenuated in ern1-1, and further declined in cyclops-3 ern1-1. ERN1 overexpression led to induction of NIN expression in cyclops-3 ern1-1 in the presence of rhizobia. Thus, in addition to CYCLOPS, ERN1 is able to increase the NIN expression level depending on infection. Furthermore, consistent with this transcriptional hierarchy, ectopic expression of ERN1 as well as NIN suppressed the IT-deficient cyclops-3 phenotype, but ERN1 failed to confer ITs in the nin-2 root. However, the ern1-1 symbiotic epidermal phenotype was not suppressed by the NIN ectopic expression. The cyclops-3 ern1-1 double mutant was less sensitive to rhizobial infection than the single mutants and defective in the symbiotic root hair response at earlier stages. This more severe phenotype of the double mutant suggests a role for ERN1 that independent of the CYCLOPS-mediated transcriptional regulation. We conclude that ERN1 is involved in regulating NIN expression in addition to CYCLOPS, and these TFs coordinately promote the symbiotic root hair response and IT development. Our data help to reveal the extensive role of ERN1 in root nodule symbiosis signaling.

Keywords

CYCLOPS ERN1 Lotus japonicus NIN Root nodule symbiosis 

Abbreviations

6-BA

6-benzylaminopurine

CCaMK

Calcium and calmodulin-dependent protein kinase

ERN1

ERF REQUIRED FOR NODULATION1

IPD3

INTERACTING PROTEIN OF DMI3

IT

INFECTION THREAD

MC

Micro-colony

NIN

NODULE INCEPTION

NFR

Nod factor receptor

NSP

NODULATION SIGNALING PATHWAY

qRT-PCR

Quantitative real-time PCR

TF

Transcription factor

WT

Wild type

Notes

Acknowledgements

We thank Dr. Emiko Yoro in Rikkyo Univeisity for her advice on the manuscript. And we thank A.T., the National Institute for Basic Biology’s Functional Genomics Facility and Model Plant Research Facility for technical supports. This work was supported by Ministry of Education, Culture, Sports, Science and Technology Grants-in-Aid for Scientific Research [Grant numbers 22128006, 17H03702 to M.K.]; and Sumitomo Foundation Grant for Basic Science Research [150574 to T.S.]. This work was also supported in part by SOKENDAI (The Graduate University for Advanced Studies).

Compliance with ethical standards

Conflict of interest

The authors have no conflicts of interest to declare.

Supplementary material

10265_2019_1122_MOESM1_ESM.pdf (583 kb)
Supplementary material 1 (PDF 582 kb)

References

  1. Amor BB, Shaw SL, Oldroyd GED, Maillet F, Penmetsa RV, Cook D et al (2003) 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.  https://doi.org/10.1046/j.1365-313X.2003.01743.x CrossRefPubMedGoogle Scholar
  2. Andriankaja WA, Boisson-Dernier A, Frances L, Sauviac L, Jauneau A, Barker DG et al (2007) AP2-ERF transcription factors mediate Nod factor-dependent MtENOD11 activation in root hairs via a novel cis-regulatory motif. Plant Cell 19:2866–2885.  https://doi.org/10.1105/tpc.107.052944 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Ardourel M, Demont N, Debellé F, Maillet F, De Billy F, Promé JC et al (1994) 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.  https://doi.org/10.1105/tpc.6.10.1357 CrossRefPubMedPubMedCentralGoogle Scholar
  4. Broughton WJ, Dilworth MJ (1971) Control of leghaemoglobin synthesis in snake beans. Biochem J 125:1075–1080.  https://doi.org/10.1042/bj1251075 CrossRefPubMedPubMedCentralGoogle Scholar
  5. Catoira R, Galera C, de Billy F, Penmetsa RV, Journet EP, Maillet F et al (2000) Four genes of Medicago truncatula controlling components of a Nod factor transduction pathway. Plant Cell 12:1647–1666.  https://doi.org/10.1105/TPC.12.9.1647 CrossRefPubMedPubMedCentralGoogle Scholar
  6. Cerri MR, Frances L, Kelner A, Fournier J, Middleton PH, Auriac MC et al (2016) The symbiosis-related ERN transcription factors act in concert to coordinate rhizobial host root infection. Plant Physiol 171:1037–1054.  https://doi.org/10.1104/pp.16.00230 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Cerri MR, Wang Q, Stolz P, Folgmann J, Frances L, Katzer K et al (2017) The ERN1 transcription factor gene is a target of the CCaMK/CYCLOPS complex and controls rhizobial infection in Lotus japonicus. New Phytol 215:323–337.  https://doi.org/10.1111/nph.14547 CrossRefPubMedGoogle Scholar
  8. Ehrhardt DW, Wais R, Long SR (1996) Calcium spiking in plant root hairs responding to rhizobium nodulation signals. Cell 85:673–681.  https://doi.org/10.1016/S0092-8674(00)81234-9 CrossRefPubMedGoogle Scholar
  9. Fisher RF, Long SR (1992) Rhizobium-plant signal exchange. Nature 357:655–660.  https://doi.org/10.1038/357655a0 CrossRefPubMedGoogle Scholar
  10. Floss DS, Gomez SK, Park HJ, MacLean AM, Müller LM, Bhattarai KK et al (2017) A transcriptional program for arbuscule degeneration during AM symbiosis is regulated by MYB1. Curr Biol 27:1206–1212.  https://doi.org/10.1016/j.cub.2017.03.003 CrossRefPubMedGoogle Scholar
  11. Fonouni-Farde C, Tan S, Baudin M, Brault M, Wen J, Mysore KS et al (2016) DELLA-mediated gibberellin signalling regulates Nod factor signalling and rhizobial infection. Nat Commun 7:12636.  https://doi.org/10.1038/ncomms12636 CrossRefPubMedPubMedCentralGoogle Scholar
  12. Guinel FC, Geil RD (2002) A model for the development of the rhizobial and arbuscular mycorrhizal symbioses in legumes and its use to understand the roles of ethylene in the establishment of these two symbioses. Can J Bot 80:695–720.  https://doi.org/10.1139/B02-066 CrossRefGoogle Scholar
  13. Hayashi T, Banba M, Shimoda Y, Kouchi H, Hayashi M, Imaizumi-Anraku H (2010) A dominant function of CCaMK in intracellular accommodation of bacterial and fungal endosymbionts. Plant J 63:141–154.  https://doi.org/10.1111/j.1365-313X.2010.04228.x CrossRefPubMedPubMedCentralGoogle Scholar
  14. Heckmann AB, Lombardo F, Miwa H, Perry JA, Bunnewell S, Parniske M et al (2006) Lotus japonicus nodulation requires two GRAS domain regulators, one of which is functionally conserved in a non-legume. Plant Physiol 142:1739–1750.  https://doi.org/10.1104/pp.106.089508 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Heckmann AB, Sandal N, Bek AS, Madsen LH, Jurkiewicz A, Nielsen MW et al (2011) Cytokinin induction of root nodule primordia in Lotus japonicus is regulated by a mechanism operating in the root cortex. Mol Plant Microbe Interact 24:1385–1395.  https://doi.org/10.1094/MPMI-05-11-0142 CrossRefPubMedGoogle Scholar
  16. Hirsch S, Kim J, Munoz A, Heckmann AB, Downie JA, Oldroyd GED (2009) GRAS proteins form a DNA binding complex to induce gene expression during nodulation signaling in Medicago truncatula. Plant Cell 21:545–557.  https://doi.org/10.1105/tpc.108.064501 CrossRefPubMedPubMedCentralGoogle Scholar
  17. Jin Y, Liu H, Luo D, Yu N, Dong W, Wang C et al (2016) DELLA proteins are common components of symbiotic rhizobial and mycorrhizal signalling pathways. Nat Commun 7:12433.  https://doi.org/10.1038/ncomms12433 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Journet EP, El-Gachtouli N, Vernoud V, de Billy F, Pichon M, Dedieu A et al (2001) Medicago truncatula ENOD11: a novel RPRP-encoding early nodulin gene expressed during mycorrhization in arbuscule-containing cells. Mol Plant Microbe Interact 14:737–748.  https://doi.org/10.1094/MPMI.2001.14.6.737 CrossRefGoogle Scholar
  19. Kaló P, Gleason C, Edwards A, Marsh J, Mitra RM, Hirsch S et al (2005) Nodulation signaling in legumes requires NSP2, a member of the GRAS family of transcriptional regulators. Science 308:1786–1789.  https://doi.org/10.1126/science.1110951 CrossRefPubMedGoogle Scholar
  20. Kawaguchi M, Imaizumi-Anraku H, Koiwa H, Niwa S, Ikuta A, Syono K et al (2002) Root, root hair, and symbiotic mutants of the model legume Lotus japonicus. Mol Plant Microbe Interact 15:17–26.  https://doi.org/10.1094/MPMI.2002.15.1.17 CrossRefPubMedGoogle Scholar
  21. Kawaharada Y, Kelly S, Nielsen MW, Hjuler CT, Gysel K, Muszyńsk A et al (2015) Receptor-mediated exopolysaccharide perception controls bacterial infection. Nature 523:308–312.  https://doi.org/10.1038/nature14611 CrossRefPubMedGoogle Scholar
  22. Kawaharada Y, Nielsen MW, Kelly S, James EK, Andersen KR, Rasmussen SR et al (2017a) Differential regulation of the Epr3 receptor coordinates membrane-restricted rhizobial colonization of root nodule primordia. Nat Commun 8:14534.  https://doi.org/10.1038/ncomms14534 CrossRefPubMedPubMedCentralGoogle Scholar
  23. Kawaharada Y, James EK, Kelly S, Sandal N, Stougaard J (2017b) The ERF Required for Nodulation1 (ERN1) transcription factor is required for infection thread formation in Lotus japonicus. Mol Plant Microbe Interact 30:194–204.  https://doi.org/10.1094/MPMI-11-16-0237-R CrossRefPubMedGoogle Scholar
  24. Lévy J, Bres C, Geurts R, Chalhoub B, Kulikova O, Duc G et al (2004) A putative Ca2+ and calmodulin-dependent protein kinase required for bacterial and fungal symbioses. Science 303:1361–1364.  https://doi.org/10.1126/science.1093038 CrossRefPubMedGoogle Scholar
  25. Limpens E, Franken C, Smit P, Willemse J, Bisseling T, Geurts R (2003) LysM domain receptor kinases regulating rhizobial Nod factor. Science 302:630–634.  https://doi.org/10.1126/science.1090074 CrossRefPubMedGoogle Scholar
  26. Liu CW, Breakspear A, Guan D, Cerri MR, Jackson K, Jiang S et al (2019) NIN acts as a network hub controlling a growth module required for rhizobial infection. Plant Physiol 179:1704–1722.  https://doi.org/10.1104/pp.18.01572 CrossRefPubMedPubMedCentralGoogle Scholar
  27. Madsen EB, Madsen LH, Radutoiu S, Olbryt M, Rakwalska M, Szczyglowski K et al (2003) A receptor kinase gene of the LysM type is involved in legume perception of rhizobial signals. Nature 425:637–640.  https://doi.org/10.1038/nature02045 CrossRefPubMedGoogle Scholar
  28. Madsen LH, Tirichine L, Jurkiewicz A, Sullivan JT, Heckmann AB, Bek AS et al (2010) The molecular network governing nodule organogenesis and infection in the model legume Lotus japonicus. Nat Commun 1:1–12.  https://doi.org/10.1038/ncomms1009 CrossRefPubMedCentralGoogle Scholar
  29. Maekawa T, Kusakabe M, Shimoda Y, Sato S, Tabata S, Murooka Y et al (2008) Polyubiquitin promoter-based binary vectors for overexpression and gene silencing in Lotus japonicus. Mol Plant Microbe Interact 21:375–382.  https://doi.org/10.1094/MPMI-21-4-0375 CrossRefPubMedGoogle Scholar
  30. Maekawa T, Maekawa-Yoshikawa M, Takeda N, Imaizumi-Anraku H, Murooka Y, Hayashi M (2009) Gibberellin controls the nodulation signaling pathway in Lotus japonicus. Plant J 58:183–194.  https://doi.org/10.1111/j.1365-313X.2008.03774.x CrossRefPubMedGoogle Scholar
  31. Małolepszy A, Mun T, Sandal N, Gupta V, Dubin M, Urbański D et al (2016) The LORE1 insertion mutant resource. Plant J 88:306–317.  https://doi.org/10.1111/tpj.13243 CrossRefPubMedGoogle Scholar
  32. Marsh JF, Rakocevic A, Mitra RM, Brocard L, Sun J, Eschstruth A et al (2007) Medicago truncatula NIN is essential for rhizobial-independent nodule organogenesis induced by autoactive calcium/calmodulin-dependent protein kinase. Plant Physiol 144:324–335.  https://doi.org/10.1104/pp.106.093021 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Messinese E, Mun JH, Yeun LH, Jayaraman D, Rougé P, Barre A et al (2007) A novel nuclear protein interacts with the symbiotic DMI3 calcium- and calmodulin-dependent protein kinase of Medicago truncatula. Mol Plant Microbe Interact 20:912–921.  https://doi.org/10.1094/MPMI-20-8-0912 CrossRefPubMedGoogle Scholar
  34. Middleton PH, Jakab J, Penmetsa RV, Starker CG, Doll J, Kaló P et al (2007) An ERF transcription factor in Medicago truncatula that is essential for Nod Factor signal transduction. Plant Cell 19:1221–1234.  https://doi.org/10.1105/tpc.106.048264 CrossRefPubMedPubMedCentralGoogle Scholar
  35. Miller DD, De Ruijter NCA, Bisseling T, Emons AMC (1999) The role of actin in root hair morphogenesis: studies with lipochito-oligosaccharide as a growth stimulator and cytochalasin as an actin perturbing drug. Plant J 17:141–154.  https://doi.org/10.1046/j.1365-313X.1999.00358.x CrossRefGoogle Scholar
  36. Miwa H, Sun J, Oldroyd GED, Downie JA (2006) Analysis of Nod-factor-induced calcium signaling in root hairs of symbiotically defective mutants of Lotus japonicus. Mol Plant Microbe Interact 19:914–923.  https://doi.org/10.1094/MPMI-19-0914 CrossRefPubMedGoogle Scholar
  37. Murakami E, Cheng J, Gysel K, Bozsoki Z, Kawaharada Y, Hjuler CT et al (2018) Epidermal LysM receptor ensures robust symbiotic signalling in Lotus japonicus. Elife 7:e33506.  https://doi.org/10.7554/eLife.33506 CrossRefPubMedPubMedCentralGoogle Scholar
  38. Murray JD, Karas BJ, Sato S, Tabata S, Amyot L, Szczyglowski K (2007) A cytokinin perception mutant colonized by rhizobium in the absence of nodule organogenesis. Science 315:101–104.  https://doi.org/10.1126/science.1132514 CrossRefPubMedGoogle Scholar
  39. Oldroyd GED (2013) Speak, friend, and enter: signalling systems that promote beneficial symbiotic associations in plants. Nat Rev Microbiol 11:252–263.  https://doi.org/10.1038/nrmicro2990 CrossRefPubMedGoogle Scholar
  40. Oldroyd GED, Long SR (2003) Identification and characterization of Nodulation-Signaling Pathway 2, a gene of Medicago truncatula involved in Nod actor signaling. Plant Physiol 131:1027–1032.  https://doi.org/10.1104/pp.102.010710 CrossRefPubMedPubMedCentralGoogle Scholar
  41. Ott T, van Dongen JT, Gunther C, Krusell L, Desbrosses G, Vigeolas H et al (2005) Symbiotic leghemoglobins are crucial for nitrogen fixation in legume root nodules but not for general plant growth and development. Curr Biol 15:531–535.  https://doi.org/10.1016/J.CUB.2005.01.042 CrossRefPubMedGoogle Scholar
  42. Pimprikar P, Carbonnel S, Paries M, Katzer K, Klingl V, Bohmer MJ et al (2016) A CCaMK-CYCLOPS-DELLA complex activates transcription of RAM1 to regulate arbuscule branching. Curr Biol 26:987–998.  https://doi.org/10.1016/j.cub.2016.01.069 CrossRefGoogle Scholar
  43. Plet J, Wasson A, Ariel F, Le Signor C, Baker D, Mathesius U et al (2011) MtCRE1-dependent cytokinin signaling integrates bacterial and plant cues to coordinate symbiotic nodule organogenesis in Medicago truncatula. Plant J 65:622–633.  https://doi.org/10.1111/j.1365-313X.2010.04447.x CrossRefPubMedGoogle Scholar
  44. Popp C, Ott T (2011) Regulation of signal transduction and bacterial infection during root nodule symbiosis. Curr Opin Plant Biol 14:458–467.  https://doi.org/10.1016/j.pbi.2011.03.016 CrossRefPubMedGoogle Scholar
  45. Qiu L, Lin J, Xu J, Sato S, Parniske M, Wang TL et al (2015) SCARN a novel class of SCAR protein that is required for root-hair infection during legume nodulation. PLoS Genet 11:e1005623.  https://doi.org/10.1371/journal.pgen.1005623 CrossRefPubMedPubMedCentralGoogle Scholar
  46. Radutoiu S, Madsen LH, Madsen EB, Felle HH, Umehara Y, Grønlund M et al (2003) Plant recognition of symbiotic bacteria requires two LysM receptor-like kinases. Nature 425:585–592.  https://doi.org/10.1038/nature02039 CrossRefPubMedGoogle Scholar
  47. Sandal N, Petersen TR, Murray J, Umehara Y, Karas B, Yano K et al (2006) Genetics of symbiosis in Lotus japonicus: recombinant inbred lines, comparative genetic maps, and map position of 35 symbiotic loci. Mol Plant Microbe Interact 19:80–91.  https://doi.org/10.1094/MPMI-19-0080 CrossRefPubMedGoogle Scholar
  48. Schauser L, Roussis A, Stiller J, Stougaard J (1999) A plant regulator controlling development of symbiotic root nodules. Nature 402:191–195.  https://doi.org/10.1038/46058 CrossRefPubMedGoogle Scholar
  49. Singh S, Katzer K, Lambert J, Cerri M, Parniske M (2014) CYCLOPS, A DNA-binding transcriptional activator, orchestrates symbiotic root nodule development. Cell Host Microbe 15:139–152.  https://doi.org/10.1016/j.chom.2014.01.011 CrossRefPubMedGoogle Scholar
  50. Smit P, Raedts J, Portyanko V, Debellé F, Gough C, Bisseling T (2005) NSP1 of the GRAS protein family is essential for rhizobial Nod factor-induced transcription. Science 308:1789–1791.  https://doi.org/10.1126/science.1111025 CrossRefPubMedGoogle Scholar
  51. Soyano T, Kouchi H, Hirota A, Hayashi M (2013) NODULE INCEPTION directly targets NF-Y subunit genes to regulate essential processes of root nodule development in Lotus japonicus. PLoS Genet 9:e1003352.  https://doi.org/10.1371/journal.pgen.1003352 CrossRefPubMedPubMedCentralGoogle Scholar
  52. Soyano T, Hirakawa H, Sato S, Hayashi M, Kawaguchi M (2014) NODULE INCEPTION creates a long-distance negative feedback loop involved in homeostatic regulation of nodule organ production. Proc Natl Acad Sci 111:14607–14612.  https://doi.org/10.1073/pnas.1412716111 CrossRefPubMedGoogle Scholar
  53. Tirichine L, Imaizumi-Anraku H, Yoshida S, Murakami Y, Madsen LH, Miwa H et al (2006) Deregulation of a Ca2+/calmodulin-dependent kinase leads to spontaneous nodule development. Nature 441:1153–1156.  https://doi.org/10.1038/nature04862 CrossRefPubMedGoogle Scholar
  54. Vernié T, Kim J, Frances L, Ding Y, Sun J, Guan D et al (2015) The NIN transcription factor coordinates diverse nodulation programs in different tissues of the Medicago truncatula root. Plant Cell 27:3410–3424.  https://doi.org/10.1105/tpc.15.00461 CrossRefPubMedPubMedCentralGoogle Scholar
  55. Xie F, Murray JD, Kim J, Heckmann AB, Edwards A, Oldroyd GED et al (2012) Legume pectate lyase required for root infection by rhizobia. Proc Natl Acad Sci USA 109:633–638.  https://doi.org/10.1073/pnas.1113992109 CrossRefPubMedGoogle Scholar
  56. Yano K, Yoshida S, Müller J, Singh S, Banba M, Vickers K et al (2008) CYCLOPS, a mediator of symbiotic intracellular accommodation. Proc Natl Acad Sci USA 105:20540–20545.  https://doi.org/10.1073/pnas.0806858105 CrossRefPubMedGoogle Scholar
  57. Yano K, Aoki S, Liu M, Umehara Y, Suganuma N, Iwasaki W et al (2017) Function and evolution of a Lotus japonicus AP2/ERF family transcription factor that is required for development of infection threads. DNA Res 24:193–203.  https://doi.org/10.1093/dnares/dsw052 CrossRefPubMedGoogle Scholar

Copyright information

© The Botanical Society of Japan and Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Meng Liu
    • 1
    • 2
  • Takashi Soyano
    • 1
    • 2
  • Koji Yano
    • 1
  • Makoto Hayashi
    • 3
  • Masayoshi Kawaguchi
    • 1
    • 2
    Email author
  1. 1.Division of Symbiotic SystemsNational Institute for Basic BiologyOkazakiJapan
  2. 2.Department of Basic Biology, School of Life ScienceSOKENDAI (The Graduate University for Advanced Studies)OkazakiJapan
  3. 3.Center for Sustainable Resource Science, RIKENYokohamaJapan

Personalised recommendations