Abstract
RNA interference (RNAi) acts through transcriptional and post-transcriptional gene silencing of homologous sequences. With the goal of using RNAi as a tool for studying gene function in the related basidiomycete cereal pathogens Ustilago hordei and Ustilago maydis, we developed a general purpose RNAi expression vector. Tandem, inverted fragments of the GUS gene were inserted into this vector flanking an intron and used to transform engineered GUS-expressing haploid cells. Down-regulation of the GUS gene and production of siRNAs were seen only in U. hordei, even though corresponding GUS double-stranded RNA was detected in both species. Similarly, when the endogenous bW mating-type gene was targeted by RNAi, mating was reduced only in U. hordei. Our work demonstrates the feasibility of using RNAi in U. hordei and provides experimental support for the observed lack of RNAi components in the U. maydis genome. We hypothesize that the sharply limited transposon complement in U. maydis is a biological consequence of this absence.
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Acknowledgments
The authors wish to thank Drs. J. Jovel, J. W. Kronstad and H. Sanfaçon for comments and valuable discussions. This research was partially funded through a grant from the Natural Sciences and Engineering Research Council of Canada to G. B.
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Communicated by B. Cormack.
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Laurie, J.D., Linning, R. & Bakkeren, G. Hallmarks of RNA silencing are found in the smut fungus Ustilago hordei but not in its close relative Ustilago maydis . Curr Genet 53, 49–58 (2008). https://doi.org/10.1007/s00294-007-0165-7
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DOI: https://doi.org/10.1007/s00294-007-0165-7