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Three endochitinase-encoding genes identified in the biocontrol fungus Clonostachys rosea are differentially expressed

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Abstract

Three endochitinase-encoding genes, cr-ech58, cr-ech42 and cr-ech37 were identified and characterised from the mycoparasitic C. rosea strain IK726. The endochitinase activity was specifically induced in media containing chitin or Fusarium culmorum cell walls as sole carbon sources. RT-PCR analysis showed that the three genes were differentially expressed. The expression of the cr-ech42 and cr-ech37 genes was triggered by F. culmorum cell walls and chitin whereas glucose repressed their expression. In contrast, the expression of cr-ech58 was not triggered by F. culmorum cell walls and chitin, suggesting a different role for this endochitinase. Phylogenetically, the cr-ech42 and cr-ech37 genes showed to be orthologous to endochitinase 42 and 37 kDa encoding genes from other mycoparasitic fungi, while no orthologous gene for the cr-ech58 gene was found. Three genetically modified mutants of C. rosea were made by disruption of the endochitinase genes via Agrobacterium-mediated transformation and their biocontrol activity was evaluated. While in planta bioassays showed no significant difference in biocontrol efficacy between the disruptants and the wildtype, the real time RT-PCR analysis showed that disruption of each endochitinase gene affected the activity of C. rosea during interaction with F. culmorum in liquid cultures.

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Acknowledgments

Financial support from Danish Research Agency, Ministry of Science Technology and Innovation (Grant No.: 23-04-0081) as well as financial support to Mojtaba Mamarabadi from the Iranian Ministry of Science, Research and Technology is gratefully acknowledged. Excellent technical support from Karin Olesen is highly appreciated.

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  1. Mette Lübeck

    Present address: Department of Life Sciences, University of Aalborg, Lautrupvang 15, 2750, Ballerup, Denmark

Authors and Affiliations

  1. Plant Pathology Section, Department of Plant Biology, Faculty of Life Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871, Frederiksberg C, Copenhagen, Denmark

    Mojtaba Mamarabadi, Birgit Jensen & Mette Lübeck

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  1. Mojtaba Mamarabadi
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  2. Birgit Jensen
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Correspondence to Mette Lübeck.

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Communicated by U. Kück.

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294_2008_199_MOESM1_ESM.doc

Supplementary material. Nucleotide and deduced amino acid sequence of cr-ech58 (A), cr-ech42 (B) and cr-ech37 (C). The nucleotide sequences are numbered on the left relative to the first nucleotide of the initiation codon, and the amino acid sequences are numbered on the right. The putative TATA box, CAAT box and poly A-tail are underlined. The putative transcription start site is in bold capital letter. The translation and stop sites are in bold italics. The cr-ech58 and cr-ech42 open reading frames were interrupted by three and two introns, respectively, which were shown with --- and the donor site, acceptor site and lariat sequence are marked with iii. The amino acid signal peptides are shown in bold letters. (DOC 23 kb)

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Mamarabadi, M., Jensen, B. & Lübeck, M. Three endochitinase-encoding genes identified in the biocontrol fungus Clonostachys rosea are differentially expressed. Curr Genet 54, 57–70 (2008). https://doi.org/10.1007/s00294-008-0199-5

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  • DOI: https://doi.org/10.1007/s00294-008-0199-5

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