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Suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi

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Citation

Gesing, S., Schindler, D., & Nowrousian, M. (2013). Suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi. JOURNAL OF BASIC MICROBIOLOGY, 53(9), 742-751. doi:10.1002/jobm.201200223.


Cite as: https://hdl.handle.net/21.11116/0000-000D-079A-E
Abstract
Ascomycetes differentiate four major morphological types of fruiting
bodies (apothecia, perithecia, pseudothecia and cleistothecia) that are
derived from an ancestral fruiting body. Thus, fruiting body
differentiation is most likely controlled by a set of common core genes.
One way to identify such genes is to search for genes with evolutionary
conserved expression patterns. Using suppression subtractive
hybridization (SSH), we selected differentially expressed transcripts in
Pyronema confluens (Pezizales) by comparing two cDNA libraries specific
for sexual and for vegetative development, respectively. The expression
patterns of selected genes from both libraries were verified by
quantitative real time PCR. Expression of several corresponding
homologous genes was found to be conserved in two members of the
Sordariales (Sordaria macrospora and Neurospora crassa), a derived group
of ascomycetes that is only distantly related to the Pezizales. Knockout
studies with N. crassa orthologues of differentially regulated genes
revealed a functional role during fruiting body development for the gene
NCU05079, encoding a putative MFS peptide transporter. These data
indicate conserved gene expression patterns and a functional role of the
corresponding genes during fruiting body development; such genes are
candidates of choice for further functional analysis.