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Published online ahead of print on 1 October 2009 as doi:10.1099/mic.0.032763-0
Microbiology (2009), DOI 10.1099/mic.0.032763-0
© 2009 Society for General Microbiology
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Microbiology 0 (2009), mic.0.032763; DOI  10.1099/mic.0.032763-0
© 2009 Society for General Microbiology

Evidence for Dicer-dependent RNA interference in the industrial penicillin producer Penicillium chrysogenum

Danielle Janus, Birgit Hoff and Ulrich Kück1

Ruhr-Universität Bochum

RNA interference (RNAi) is a sequence-specific post-transcriptional gene silencing system that downregulates target gene expression. Here, we provide several lines of evidence for RNA silencing in the industrial β-lactam antibiotic producer Penicillium chrysogenum using the DsRed reporter gene under the control of the constitutive trpC promoter or the inducible xylP promoter. The functional RNAi system was verified by detection of siRNAs that hybridize exclusively with gene-specific 32P-labeled RNA probes. Moreover, when RNAi was used to silence the endogenous PcbrlA morphogene that controls fungal conidiophore development, a dramatic reduction in the formation of conidiospores was observed in 47 % of the corresponding transformants. Finally, evidence that RNAi in P. chrysogenum is dependent on a Dicer peptide was provided with a strain lacking the Pcdcl2. In the {Delta}Pcdcl2 background, silencing of the PcbrlA gene, controlling conidiophores development, was tested. None of the analyzed transformants showed a developmental defect. The applicability of the RNAi system in P. chrysogenum was finally demonstrated by silencing the Pcku70 gene to increase homologous recombination frequency. This led to the generation of single and double knockout mutants.

1 E-mail: ulrich.kueck{at}rub.de






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