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Cross-species microarray hybridization to identify developmentally regulated genes in the filamentous fungus Sordaria macrospora

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Abstract

The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies that protect the developing ascospores and ensure their proper discharge. Several regulatory genes essential for fruiting body development were previously isolated by complementation of the sterile mutants pro1, pro11 and pro22. To establish the genetic relationships between these genes and to identify downstream targets, we have conducted cross-species microarray hybridizations using cDNA arrays derived from the closely related fungus Neurospora crassa and RNA probes prepared from wild-type S. macrospora and the three developmental mutants. Of the 1,420 genes which gave a signal with the probes from all the strains used, 172 (12%) were regulated differently in at least one of the three mutants compared to the wild type, and 17 (1.2%) were regulated differently in all three mutant strains. Microarray data were verified by Northern analysis or quantitative real time PCR. Among the genes that are up- or down-regulated in the mutant strains are genes encoding the pheromone precursors, enzymes involved in melanin biosynthesis and a lectin-like protein. Analysis of gene expression in double mutants revealed a complex network of interaction between the pro gene products.

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Acknowledgements

The authors thank Swenja Ellßel and Susanne Schlewinski for excellent technical assistance, Dr. Stefanie Pöggeler for primers and probes for the ppg genes, and Dr. Georg Zoidl (Bochum) for generous provision of real time PCR facilities. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 480 project A1)

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Authors and Affiliations

  1. Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum ND 7/131, Universitätsstr. 150, 44801, Bochum, Germany

    Minou Nowrousian & Ulrich Kück

  2. Departments of Genetics and Biochemistry, Dartmouth Medical School, Dartmouth, NH, 03755, USA

    Carol Ringelberg, Jay C. Dunlap & Jennifer J. Loros

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  1. Minou Nowrousian
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  2. Carol Ringelberg
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  5. Ulrich Kück
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Correspondence to Ulrich Kück.

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Communicated by P.J. Punt

Note added in proof

Note added in proof

While our manuscript was under review, Te Biesebeke and coworkers described cross-species hybridization of cDNA macroarrays from the filamentous fungus Aspergillus niger with Aspergillus oryzae targets (Te Biesebeke et al. 2005, Mol Genet Genomics, in press). Our finding that pgg transcript levels are up-regulated in the mutants pro1, pro11, and pro22 was recently supported by an investigation demonstrating that the pheromones themselves are up-regulated in the mutant strains (Mayrhofer and Pöggeler 2005, Eukaryot Cell, in press).

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Nowrousian, M., Ringelberg, C., Dunlap, J.C. et al. Cross-species microarray hybridization to identify developmentally regulated genes in the filamentous fungus Sordaria macrospora. Mol Genet Genomics 273, 137–149 (2005). https://doi.org/10.1007/s00438-005-1118-9

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  • DOI: https://doi.org/10.1007/s00438-005-1118-9

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