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Determining Genetic Expression Profiles in C. elegans Using Microarray and Real-time PCR
Published on: July 30, 2011
Microarray and real-time PCR analyses reveal mating type-dependent gene expression in a homothallic fungus
S Pöggeler1, M Nowrousian, C Ringelberg
1Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, 44780, Bochum, Germany. stefanie.poeggeler@rub.de
Abstract:
Sordaria macrospora is a homothallic ascomycete which is able to form fertile fruiting bodies without a mating partner. To analyze the molecular basis of homothallism and the role of mating products during fruiting body development, we have deleted the mating type gene Smta-1 encoding a high-mobility group domain (HMG) protein. The DeltaSmta-1 deletion strain is morphologically wild type during vegetative growth, but it is unable to produce perithecia or ascospores. To identify genes expressed under control of Smta-1, we performed a cross-species microarray analysis using Neurospora crassa cDNA microarrays hybridized with S. macrospora targets. We identified 107 genes that are more than twofold up- or down-regulated in the mutant. Functional classification revealed that 81 genes have homologues with known or putative functions. Comparison of array data from DeltaSmta-1 with those from three phenotypically similar mutants revealed that only a limited set of ten genes is deregulated in all mutants. Remarkably, the ppg2 gene encoding a putative lipopeptide pheromone is 500-fold down-regulated in the DeltaSmta-1 mutant while in all other sterile mutants this gene is up-regulated.
Insights
Sordaria macrospora
Area of Science:
- Mycology
- Molecular Biology
- Genetics
Background:
- Sordaria macrospora is a homothallic ascomycete capable of self-fertilization.
- Understanding homothallism requires analyzing mating type genes and their regulatory roles.
- The mating type gene Smta-1 encodes a high-mobility group (HMG) domain protein crucial for sexual development.
Purpose of the Study:
- To investigate the molecular mechanisms underlying homothallism in Sordaria macrospora.
- To determine the role of the mating type gene Smta-1 in fruiting body development.
- To identify genes regulated by Smta-1 during sexual reproduction.
Main Methods:
- Deletion of the mating type gene Smta-1 to create a DeltaSmta-1 mutant strain.
- Cross-species microarray analysis using Neurospora crassa cDNA microarrays.
- Hybridization of S. macrospora targets to identify differentially expressed genes.
Main Results:
- The DeltaSmta-1 deletion strain exhibited normal vegetative growth but failed to produce perithecia or ascospores.
- Microarray analysis identified 107 genes significantly up- or down-regulated in the mutant.
- The ppg2 gene, encoding a putative lipopeptide pheromone, was found to be 500-fold downregulated in the DeltaSmta-1 mutant, contrasting with its upregulation in other sterile mutants.
Conclusions:
- Smta-1 is essential for sexual development and fertility in Sordaria macrospora.
- Smta-1 regulates a suite of genes involved in sexual reproduction.
- The unique regulation of ppg2 in the DeltaSmta-1 mutant suggests a specific role for Smta-1 in pheromone pathway control.
