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Gene expression in mycorrhizal orchid protocorms suggests a friendly plant–fungus relationship

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Abstract

Main conclusion

Orchid mycorrhiza has been often interpreted as an antagonistic relationship. Our data on mycorrhizal protocorms do not support this view as plant defence genes were not induced, whereas some nodulin-like genes were significantly up-regulated.

Orchids fully depend on symbiotic interactions with specific soil fungi for seed germination and early development. Germinated seeds give rise to a protocorm, a heterotrophic organ that acquires nutrients, including organic carbon, from the mycorrhizal partner. It has long been debated if this interaction is mutualistic or antagonistic. To investigate the molecular bases of the orchid response to mycorrhizal invasion, we developed a symbiotic in vitro system between Serapias vomeracea, a Mediterranean green meadow orchid, and the rhizoctonia-like fungus Tulasnella calospora. 454 pyrosequencing was used to generate an inventory of plant and fungal genes expressed in mycorrhizal protocorms, and plant genes could be reliably identified with a customized bioinformatic pipeline. A small panel of plant genes was selected and expression was assessed by real-time quantitative PCR in mycorrhizal and non-mycorrhizal protocorm tissues. Among these genes were some markers of mutualistic (e.g. nodulins) as well as antagonistic (e.g. pathogenesis-related and wound/stress-induced) genes. None of the pathogenesis or wound/stress-related genes were significantly up-regulated in mycorrhizal tissues, suggesting that fungal colonization does not trigger strong plant defence responses. In addition, the highest expression fold change in mycorrhizal tissues was found for a nodulin-like gene similar to the plastocyanin domain-containing ENOD55. Another nodulin-like gene significantly more expressed in the symbiotic tissues of mycorrhizal protocorms was similar to a sugar transporter of the SWEET family. Two genes coding for mannose-binding lectins were significantly up-regulated in the presence of the mycorrhizal fungus, but their role in the symbiosis is unclear.

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Abbreviations

EST:

Expressed sequence tag

GO:

Gene ontology

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Acknowledgments

We thank Emilie Tisserant (INRA, France) for help with MIRA assembly of the transcriptomic sequences. MiR was supported by the Progetto Lagrange (Fondazione CRT), FS and EE by PhD MIUR fellowships. We acknowledge partial funding by the Italian MIUR (PRIN2007), IPP-CNR and local funding by University of Turin. The UMR1136 is supported by a grant overseen by the French National Research Agency (ANR) as part of the: “Investissements d’Avenir” program (ANR-11-LABX-0002-01, Lab of Excellence ARBRE).

Conflict of interest

The authors declare that they have no conflict of interest.

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Author notes

  1. Michele Rodda

    Present address: Singapore Botanic Gardens Herbarium, 1 Cluny Road, Singapore, 259569, Singapore

Authors and Affiliations

  1. Department of Life Sciences and Systems Biology, University of Turin, Viale Mattioli 25, 10125, Turin, Italy

    Silvia Perotto, Marco Rodda, Alex Benetti, Fabiano Sillo, Enrico Ercole, Michele Rodda & Mariangela Girlanda

  2. Institute for Plant Protection-CNR, Viale Mattioli 25, 10125, Turin, Italy

    Silvia Perotto, Mariangela Girlanda & Raffaella Balestrini

  3. UMR1136 INRA/Université de Lorraine, “Interactions Arbres/Micro-organismes”, Lab of Excellence ARBRE, INRA, 54280, Nancy, Champenoux, France

    Claude Murat

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  1. Silvia Perotto
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Correspondence to Silvia Perotto.

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425_2014_2062_MOESM1_ESM.tif

Supplementary material 1 (TIFF 3174 kb) Fig. S1 Gene Ontology assignments for S. vomeracea transcripts. Level 3 annotations are shown for the biological process and cellular component graphs, and level 2 annotations for the molecular function graph

425_2014_2062_MOESM2_ESM.tif

Supplementary material 2 (TIFF 3174 kb) Fig. S2 Phylogenetic relationships of plant serine carboxipeptidases-like (SCPL) proteins based on amino acid deduced sequences. The sequences were aligned using Muscle and the unrooted tree was constructed using maximum likelihood. Characterized SCPL with acyltransferase activity from Arabidopsis and Oryza sativa are included in the tree and form a separate cluster. Characterized SCPL acyltransferase enzymes included in the tree are also from Avena sativa (ACT21078_SCPL1) and Brassica napus (AAQ91191_SCT). Characterized serine carboxypeptidase sequences from Hordeum vulgare were included (Hv, P07519_SCPI and P2159.2_SCPIII), in addition to sequences from Vitis vinifera (Vv, XP_002264454), Ricinus communis (Rc, XP_002527263) and Medicago truncatula MtSCP1 (Mt, XP_003592239). Numbers indicate bootstrap values and are indicated only when ≥ 50 %. Arrows point to the S. vomeracea sequence (SvWound3)

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Perotto, S., Rodda, M., Benetti, A. et al. Gene expression in mycorrhizal orchid protocorms suggests a friendly plant–fungus relationship. Planta 239, 1337–1349 (2014). https://doi.org/10.1007/s00425-014-2062-x

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