Skip to main content
SpringerLink
Log in

Medicago truncatula syntaxin SYP132 defines the symbiosome membrane and infection droplet membrane in root nodules

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Symbiotic association of legume plants with rhizobia bacteria culminates in organogenesis of nitrogen-fixing root nodules. In indeterminate nodules, plant cells accommodate rhizobial infection by enclosing each bacterium in a membrane-bound, organelle-like compartment called the symbiosome. Numerous symbiosomes occupy each nodule cell; therefore an enormous amount of membrane material must be delivered to the symbiosome membrane for its development and maintenance. Protein delivery to the symbiosome is thought to rely on the plant secretory system; however, the targeting mechanisms are not well understood. In this study, we report the first in-depth analysis of a syntaxin localized on symbiosome membranes. Syntaxins help define a biochemical identity to each compartment in the plant secretory system and facilitate vesicle docking and fusion. Here, we present biochemical and cytological evidence that the SNARE MtSYP132, a Medicago truncatula homologue of Arabidopsis thaliana Syntaxin of Plants 132, localizes to the symbiosome membrane. Using a specific anti-MtSYP132 peptide antibody, we also show that MtSYP132 localizes to the plasma membrane surrounding infection threads and is most abundant on the infection droplet membrane. These results indicate that MtSYP132 may function in infection thread development or growth and the early stages of symbiosome formation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ELISA:

Enzyme-linked immunosorbent assay

Fmoc:

9-Flourenylmethloxycarbonyl

KLH:

Keyhole limpet hemocyanin

MtSYP132:

Medicago truncatula syntaxin

Pipes:

Piperazine-1,4-bis(2-ethanesulfonic acid)

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

SNARE:

Soluble N-ethylmaleimide-sensitive factor adaptor protein receptors

TBS:

Tris buffered saline

TBST:

TBS with Tween-20

References

  • Aalto MK, Ronne H, Keranen S (1993) Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport. EMBO J 12:4095–4104

    PubMed  CAS  Google Scholar 

  • Altschul SF, Madden TL, Schdffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  PubMed  CAS  Google Scholar 

  • Assaad FF, Huet Y, Mayer U, Jurgens G (2001) The cytokinesis gene Keule encodes a Sec1 protein that binds the syntaxin KNOLLE. J Cell Biol 152:531–543

    Article  PubMed  CAS  Google Scholar 

  • Assaad FF, Qiu J, Youngs H, Ehrhardt D, Zimmerli L, Kalde M, Wanner G, Peck SC, Edwards H, Ramonell K, Sommerville CR, Thordal-Christensen H (2004) The PEN1 syntaxin defines a novel cellular compartment upon fungal attack and is required for the timely assembly of papillae. Mol Biol Cell 15:5118–5129

    Article  PubMed  CAS  Google Scholar 

  • Bassham DC, Sanderfoot AA, Kovaleva V, Zheng H, Raikhel NV (2000) AtVPS45 complex formation at the trans-Golgi network. Mol Biol Cell 11:2251–2265

    PubMed  CAS  Google Scholar 

  • Bennett MK, Garcia-Arras JE, Elferink K, Peterson K, Fleming AM, Hazuka CD, Scheller RH (1993) The syntaxin family of vesicular transport receptors. Cell 74:863–873

    Article  PubMed  CAS  Google Scholar 

  • Blatt MR, Leyman B, Geelen D (1999) Tansley review no. 108: molecular events of vesicle trafficking and control by SNARE proteins in plants. New Phytol 144:389–418

    Article  CAS  Google Scholar 

  • Brewin N (2004) Plant cell wall remodelling in the Rhizobium–legume symbiosis. Crit Rev Plant Sci 234:293–316

    Article  CAS  Google Scholar 

  • Brewin N, Wood EA, Bradley DJ, Harding SC, Sindhu SS, Kannenberg EL, VandenBosch KA (1988) The use of monoclonal antibodies to study plant–microbe interactions in the pea nodule. In: Bothe H, de Bruijn FJ, Newton WE (eds) Nitrogen fixation: hundred years after. Gustav Fischer Verlag, Stuttgart, pp 517–525

    Google Scholar 

  • Carter CJ, Bednarek SY, Raikhel NV (2004) Membrane trafficking in plants: new discoveries and approaches. Curr Opin Plant Biol 7:701–707

    Article  PubMed  CAS  Google Scholar 

  • Catalano CM, Lane WS, Sherrier DJ (2004) Biochemical characterization of symbiosome membrane proteins from Medicago truncatula root nodules. Electrophoresis 25:519–531

    Article  PubMed  CAS  Google Scholar 

  • Collins RF, Schreiber AD, Grinstein S, Trimble WS (2002) Syntaxins 13 and 7 function at distinct steps during phagocytosis. J Immunol 169:3250–3256

    PubMed  CAS  Google Scholar 

  • Collins NC, Thordal-Christensen H, Lipka V, Bau S, Kombrink E, Qiu JL, Huckelhoven R, Stein M, Freialdenhoven H, Somerville SC, Schulze-Lefert P (2003) SNARE-protein-mediated disease resistance at the plant cell wall. Nature 425:973–977

    Article  PubMed  CAS  Google Scholar 

  • Gage DJ (2004) Infection and invasion of roots by symbiotic nitrogen-fixing rhizobia during nodulation of temperate legumes. Microbiol Mol Biol Rev 68:280–300

    Article  PubMed  CAS  Google Scholar 

  • Gasteiger E, Gattiker A, Hoogland C, Ivanyi I, Appel RD, Bairoch A (2003) ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Res 31:3784–3788

    Article  PubMed  CAS  Google Scholar 

  • Hackman DJ, Rotstein OD, Bennett MK, Klip A, Grinstein S, Manolson MF (1996) Characterization and subcellular localization of target membrane soluble NSF attachment protein receptors (t-SNAREs) in macrophages. Syntaxins 2, 3, and 4 are present on phagosomal membranes. J Immunol 156:4377–4383

    Google Scholar 

  • Haynes JG, Czymmek KJ, Carlson CA, Veershlingam H, Dickstein R, Sherrier DJ (2004) Rapid analysis of legume root nodule development using confocal microscopy. New Phytol 163:661–668

    Article  Google Scholar 

  • Homann U (1998) Fusion and fission of plasma membrane material accommodates for osmotically induced changes in surface area in guard-cell protoplasts. Planta 206:329–333

    Article  CAS  Google Scholar 

  • Jahn R, Lang T, Südhof TC (2003) Membrane fusion. Cell 112:519–533

    Article  PubMed  CAS  Google Scholar 

  • Lauber MH, Waizenegger I, Steinmann T, Schwarz H, Mayer U, Hwang I, Lukowitz W, Jurgens G (1997) The Arabidopsis KNOLLE protein is a cytokinesis-specific syntaxin. J Cell Biol 139:1485–1493

    Article  PubMed  CAS  Google Scholar 

  • Lee Y, Tsai J, Sunkara S, Karamycheva S, Pertea G, Sultana R, Antonescu V, Chan A, Cheung F, Quackenbush J (2005) The TIGR Gene Indices: clustering and assembling EST and known genes and integration with eukaryotic genomes. Nucleic Acids Res 33(Database Issue): D41–D44

    Google Scholar 

  • Leyman B, Geelen D, Quintero FJ, Blatt MR (1999) A tobacco syntaxin with a role in hormonal control of guard cell ion channels. Science 283:537–540

    Article  PubMed  CAS  Google Scholar 

  • McNew JA, Parlati F, Fukuda R, Johnston J, Paz K, Paumet F, Sollner TH, Rothman JH (2000) Compartmental specificity of cellular membrane fusion encoded in SNARE proteins. Nature 407:153–159

    Article  PubMed  CAS  Google Scholar 

  • Meade HM, Long SR, Ruvkun GB, Brown SE, Ausubel FM (1982) Physical and genetic characterization of symbiotic and auxotropic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol 149:114–122

    PubMed  CAS  Google Scholar 

  • Muller I, Wagner W, Volker A, Schellmann S, Nacry P, Kuttner F, Schwarz-Sommer Z, Mayer U, Jurgens G (2003) Syntaxin specificity of cytokinesis in Arabidopsis. Nature Cell Biol 5:531–534

    Article  PubMed  CAS  Google Scholar 

  • Pratelli R, Sutter J, Blatt MR (2004) A new catch in the SNARE. Trends Plant Sci 9:187–195

    Article  PubMed  CAS  Google Scholar 

  • Rae A, Bonfante-Fasolo P, Brewin N (1992) Structure and growth of infection threads in the legume symbiosis with Rhizobium leguminosarum. Plant J 2:385–395

    Article  Google Scholar 

  • Reed NA, Oh DJ, Czymmek K, Duncan MK (2001) An immunohistochemical method for the detection of proteins in the vertebrate lens. J Immunol Methods 253:243–252

    Article  PubMed  CAS  Google Scholar 

  • Rojo E, Zouhar J, Kovaleva V, Hong S, Raikhel NV (2003) The AtC-VPS protein complex is localized to the tonoplast and the prevacuolar compartment in Arabidopsis. Mol Biol Cell 14:361–369

    Article  PubMed  CAS  Google Scholar 

  • Roth E, Jeon K, Stacey G (1988) Homology in endosymbiotic systems: the term “symbiosome”. In: Palacios R, Verma DPS (eds) Molecular Genetics of Plant Microbe Interactions. Am Phytopathological Soc, St. Paul, pp 220–229

    Google Scholar 

  • Sanderfoot AA, Raikhel NV (1999) The specificity of vesicle trafficking: coat proteins and SNAREs. Plant Cell 11:629–641

    Article  PubMed  CAS  Google Scholar 

  • Sanderfoot AA, Assaad F, Raikhel N (2000) The Arabidopsis Genome. An abundance of soluble N-ethylmaleimide-sensitive factor adaptor protein receptors. Plant Physiol 124:1558–1569

    Article  PubMed  CAS  Google Scholar 

  • Sanderfoot AA, Pilgrim M, Adam L, Raikhel NV (2001) Disruption of individual members of Arabidopsis syntaxin gene families indicates each has essential functions. Plant Cell 13:659–666

    Article  PubMed  CAS  Google Scholar 

  • Sato MH, Nakamura N, Ohsumi Y, Kouchi H, Kondo M, Hara-Nishimura I, Nishimura M, Wada Y (1997) The AtVAM3 encodes a syntaxin-related molecule implicated in the vacuolar assembly in Arabidopsis thaliana. J Biol Chem 272:24530–24535

    Article  PubMed  CAS  Google Scholar 

  • Surpin M, Raikhel N (2004) Traffic jams affect plant development and signal transduction. Nat Rev 5:100–109

    Article  CAS  Google Scholar 

  • Thiel G, Battey N (1998) Exocytosis in plants. Plant Mol Biol 38:111–125

    Article  PubMed  CAS  Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Article  PubMed  CAS  Google Scholar 

  • VandenBosch KA, Bradley DJ, Knox JP, Perotto S, Butcher GW, Brewin NJ (1989) Common components of the infection thread matrix and the intercellular space identified by immunocytochemical analysis of pea nodules and uninfected root. EMBO J 9:335–342

    Google Scholar 

  • Wolf E, Kim PS, Berger B (1997) Multicoil: a program for predicting two and three stranded coiled coils. Protein Sci 6:1179–1189

    Article  PubMed  CAS  Google Scholar 

  • Zhu J, Gong Z, Zhang C, Song CP, Damsz B, Inan G, Koiwa H, Zhu JK, Hasegawa PM, Bressan RA (2002) OSM1/SYP61: a syntaxin protein in Arabidopsis controls abscisic acid-mediated and non-abscisic acid-mediated responses to abiotic stress. Plant Cell 14:3009–3028

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The MS/MS spectra identified for MtSYP132 in Supplemental Figure 1 are provided courtesy of William S. Lane, Harvard University Microchemistry and Proteomics Analysis Facility. This work was supported by USDA NRI CGP awards 2005-35318-16215, 2001-35318-10915, and 2001-35311-10161 to DJS.

Author information

Authors and Affiliations

  1. Department of Plant and Soil Sciences, University of Delaware, Newark, DE, 19717, USA

    Christina M. Catalano, Janine G. Gann & D. Janine Sherrier

  2. Delaware Biotechnology Institute, University of Delaware, 15 Innovation Way, Rm 129, Newark, DE, 19707, USA

    Christina M. Catalano, Kirk J. Czymmek, Janine G. Gann & D. Janine Sherrier

  3. Department of Biological Sciences, University of Delaware, Newark, DE, 19716, USA

    Kirk J. Czymmek & D. Janine Sherrier

Authors
  1. Christina M. Catalano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kirk J. Czymmek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Janine G. Gann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D. Janine Sherrier
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to D. Janine Sherrier.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Cite this article

Catalano, C.M., Czymmek, K.J., Gann, J.G. et al. Medicago truncatula syntaxin SYP132 defines the symbiosome membrane and infection droplet membrane in root nodules. Planta 225, 541–550 (2007). https://doi.org/10.1007/s00425-006-0369-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-006-0369-y

Keywords

Search

Navigation