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Two genes encoding different truncated hemoglobins are regulated during root nodule and arbuscular mycorrhiza symbioses of Medicago truncatula

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Abstract

The MtTrHb1 and MtTrHb2 genes of the model legume Medicago truncatula Gaertn. encode proteins homologous to truncated hemoglobins (TrHb) from plants and a range of different microorganisms. Induction of MtTrHb1 in root nodules and expression of MtTrHb2 in root nodules, as well as in mycorrhizal roots, were shown by quantitative real-time reverse transcription–polymerase chain reaction (RT–PCR). The promoters of both genes were PCR-amplified and fused to the gusAint coding region. By analysing these gusAint-fusions in transgenic root tissues, we were able to localize their activity in root nodules and in roots colonized by arbuscular mycorrhizal (AM) fungi. Whereas the promoter of MtTrHb1 was activated in the infected cells of the nitrogen-fixing zone of root nodules, the MtTrHb2 promoter was predominantly active in the nodule vascular tissue. This expression pattern correlates with the presence of an ‘organ-specific element’ (OSE)-like sequence in the MtTrHb1 promoter, which is not present in the MtTrHb2 regulatory unit. Concerning the AM symbiosis, only the MtTrHb2 promoter mediated an expression in arbuscule-containing cells and in the root vascular tissue of mycorrhizal root segments colonized by the fungus Glomus intraradices.

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Abbreviations

AM :

Arbuscular mycorrhizal

BAC :

Bacterial artificial chromosome

dpi :

Days post inoculation

EST :

Expressed sequence tag

Hb :

Hemoglobin

Lb :

Leghemoglobin

NO :

Nitric oxide

OSE :

Organ-specific element

TC :

Tentative consensus sequence

TrHb :

Truncated hemoglobin

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Acknowledgements

We thank the Deutsche Forschungsgemeinschaft for supporting this work in the frame of the SPP1084 (MolMyk) project Pe814/1-2 and Prof. Alfred Pühler for critical reading of the manuscript.

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Authors and Affiliations

  1. Fakultät für Biologie, Lehrstuhl für Genetik, Universität Bielefeld, P.O. Box 100131, 33501, Bielefeld, Germany

    Martin F. Vieweg, Natalija Hohnjec & Helge Küster

  2. International NRW Graduate School in Bioinformatics and Genome Research, Center for Biotechnology, Universität Bielefeld, 33594, Bielefeld, Germany

    Natalija Hohnjec & Helge Küster

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  1. Martin F. Vieweg
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  2. Natalija Hohnjec
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  3. Helge Küster
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Correspondence to Helge Küster.

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Vieweg, M.F., Hohnjec, N. & Küster, H. Two genes encoding different truncated hemoglobins are regulated during root nodule and arbuscular mycorrhiza symbioses of Medicago truncatula. Planta 220, 757–766 (2005). https://doi.org/10.1007/s00425-004-1397-0

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