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The role of the glyoxylate cycle in the symbiotic fungus Tuber borchii: expression analysis and subcellular localization

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Abstract

Expression profiles of isocitrate lyase (TbICL), malate synthase (TbMLS) and fructose-1,6-bisphosphatase (TbFBP) from the mycorrhizal ascomycete Tuber borchii were investigated by real-time RT-PCR in fruiting bodies at different stages of maturation. In addition, a time course experiment was set up to determine how the transcription profile of TbICL, TbMLS and TbFBP in axenic-grown mycelia is affected by different carbon sources. The transcript levels of the three genes in the fruiting bodies were all much higher than those measured in the vegetative stage. The investigation on axenic-grown mycelia revealed that the main positive regulator of TbICL and TbMLS gene expression is the availability of acetate and ethanol, while oleic acid is a too complex substrate for the limited degradative capacities of T. borchii. Immunolabelling on axenic-grown mycelia showed a co-localization of TbICL and the peroxisomal marker protein FOX2. This result demonstrated that in T. borchii ICL is compartmentalized in peroxisomes. The high induction of TbICL, TbMLS and TbFBP transcription and the translocation of lipids in fruiting bodies let us hypothesize that glyoxylate cycle and gluconeogenesis are key metabolic pathways in the recycling of existing cell material and the channelling towards the biosynthesis of new cell components during the maturation of fruiting bodies.

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Abbreviations

GLOX:

Glyoxylate

ICL:

Isocitrate lyase

MLS:

Malate synthase

FBP:

Fructose -1, 6- bisphosphatase

EST:

Expressed sequence tag

CM:

Control medium

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Acknowledgments

This work was supported by grants from the Regione Piemonte (CIPE B63), IPP-CNR, Compagnia di San Paolo and CEBIOVEM (D.M. 17/10/2003 prot. n. 193/2003). The authors would like to thank Marta Vallino, Luisa Lanfranco, Gilda Cappellazzo and Laura Miozzi (Dipartimento di Biologia Vegetale dell’Università di Torino, Italy) for the helpful discussions about Real-Time RT-PCR data handling and Andrea Genre (Laboratory of Advanced Microscopy) for the support during confocal observations.

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Authors and Affiliations

  1. Dipartimento di Biologia Vegetale, Università di Torino and Istituto per la Protezione delle Piante del CNR-Sezione di Torino, Viale Mattioli 25, 10125, Turin, Italy

    Simona Abba’, Raffaella Balestrini, Alessandra Benedetto & Paola Bonfante

  2. Inst. f. Physiologische Chemie, Universitatsstr. 150, Bochum, 44780, Germany

    Hanspeter Rottensteiner

  3. Departamento de Microbiologia y Parasitologia, Fac. Farmacia, Campus Universitario, Universidad de Alcala, Alcala de Henares, Madrid, Spain

    José Ramón De Lucas

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  1. Simona Abba’
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  2. Raffaella Balestrini
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  3. Alessandra Benedetto
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  4. Hanspeter Rottensteiner
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  5. José Ramón De Lucas
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  6. Paola Bonfante
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Correspondence to Paola Bonfante.

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Communicated by U. Kües.

Simona Abba’ and Raffaella Balestrini contributed equally to this work.

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Abba’, S., Balestrini, R., Benedetto, A. et al. The role of the glyoxylate cycle in the symbiotic fungus Tuber borchii: expression analysis and subcellular localization. Curr Genet 52, 159–170 (2007). https://doi.org/10.1007/s00294-007-0149-7

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