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Comparative analysis of remodelling of the plant–microbe interface in Pisum sativum and Medicago truncatula symbiotic nodules

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Abstract

Infection of host cells by nitrogen-fixing soil bacteria, known as rhizobia, involves the progressive remodelling of the plant–microbe interface. This process was examined by using monoclonal antibodies to study the subcellular localisation of pectins and arabinogalactan proteins (AGPs) in wild-type and ineffective nodules of Pisum sativum and Medicago truncatula. The highly methylesterified homogalacturonan (HG), detected by monoclonal antibody JIM7, showed a uniform localisation in the cell wall, regardless of the cell type in nodules of P. sativum and M. truncatula. Low methylesterified HG, recognised by JIM5, was detected mainly in the walls of infection threads in nodules of both species. The galactan side chain of rhamnogalacturonan I (RG-I), recognised by LM5, was present in the nodule meristem in both species and in the infection thread walls in P. sativum, but not in M. truncatula. The membrane-anchored AGP recognised by JIM1 was observed on the plasma membrane in nodules of P. sativum and M. truncatula. In P. sativum, the AGP epitope recognised by JIM1 was present on mature symbiosome membranes of wild-type nodules, but JIM1 labelling was absent from symbiosome membranes in the mutant Sprint-2Fix− (sym31) with undifferentiated bacteroids, suggesting a possible involvement of AGP in the maturation of symbiosomes. Thus, the common and species-specific traits of cell wall remodelling during nodule differentiation were demonstrated.

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Abbreviations

MAb:

Monoclonal antibody

HG:

Homogalacturonan

RG-I:

Rhamnogalacturonan I

AGP:

Arabinogalactan protein

PBS:

Phosphate-buffered saline

BSA-C:

Acetylated bovine serum albumin

DAI:

Days after inoculation

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Acknowledgements

We thank Jean-Marie Prosperi (INRA, Montpellier, France) for providing M. truncatula seeds, Peter Mergaert (Institut des Sciences Végétal, Gif sur Yvette, France) for the kind gift of seeds of the M. truncatula dnf1–1 mutant line and Pascal Gamas (Laboratoire des Interactions Plantes-Microorganismes, Castanet-Tolosan, France) for the kind gift of seeds TR3 and efd–1 mutants. The research was performed using equipment of the Core Centrum “Genomic Technologies, Proteomics and Cell Biology” in ARRIAM and the “Molecular and Cell Technologies” Research Resource Centre at Saint-Petersburg State University.

Funding

This work was financially supported by Russian Science Foundation (16–16–10035).

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Authors and Affiliations

  1. All-Russia Research Institute for Agricultural Microbiology (ARRIAM), Laboratory of Molecular and Cellular Biology, Podbelsky chaussee 3, St.-Petersburg, Russia, 196608

    Anna V. Tsyganova, Elena V. Seliverstova & Viktor E. Tsyganov

  2. Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences, pr. Torez 44, St.-Petersburg, Russia, 194223

    Elena V. Seliverstova

  3. John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, UK

    Nicholas J. Brewin

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  1. Anna V. Tsyganova
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  2. Elena V. Seliverstova
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  4. Viktor E. Tsyganov
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Correspondence to Viktor E. Tsyganov.

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Tsyganova, A.V., Seliverstova, E.V., Brewin, N.J. et al. Comparative analysis of remodelling of the plant–microbe interface in Pisum sativum and Medicago truncatula symbiotic nodules. Protoplasma 256, 983–996 (2019). https://doi.org/10.1007/s00709-019-01355-5

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