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RNAi-mediated down-regulation of a melanin polyketide synthase (pks1) gene in the fungus Slafractonia leguminicola

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Abstract

The fungus Slafractonia leguminicola, the causal agent of blackpatch disease of legumes produces two mycotoxins slaframine and swainsonine, causing slobbers’ symptoms and locoism of grazing animals, respectively. The genetics of this important fungus is poorly understood. This work aimed to develop a genetic transformation system and evaluate the efficacy of RNA interference (RNAi) in S. leguminicola. In this study, S. leguminicola was transformed using a PEG-mediated method with a fungal construct that carries a hygromycin resistance cassette. To assess the use of RNAi, a silencing construct pSilentPKS1-AS was constructed which includes inverted repeat transgenes of the polyketide synthase gene (pks1) that is involved in melanin biosynthesis. Transformation of S. leguminicola with the IRT pks1 vector decreased pks1 transcripts levels 82–92% in knockdown mutants when compared with the wild type and was accompanied with a reduction in melanin and swainsonine production. These results demonstrate that RNAi can be a useful tool for studying gene function in S. leguminicola.

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Acknowledgements

This work was financially supported by the New Mexico State University Agricultural Experiment Station and the Molecular Biology Graduate Program. The authors thank the Fungal Genetics Stock Center for kindly providing the fungal silencing vector pSilent-1.

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Author notes

  1. Mohammad S. Alhawatema

    Present address: Tafila Technical University, Tafilah, Jordan

Authors and Affiliations

  1. Molecular Biology Graduate Program, New Mexico State University, Las Cruces, NM, 88003, USA

    Mohammad S. Alhawatema & Rebecca Creamer

  2. Horticulture Department, Sohag University, Sohag, Egypt

    Sayed Gebril

  3. U.S. Department of Agriculture, Agricultural Research Service, Poisonous Plant Research Laboratory, 1150 East 1400 North, Logan, UT, 84341, USA

    Daniel Cook

  4. Entomology, Plant Pathology, Weed Science, New Mexico State University, Box 30003, MSC 3BE, 945 College Ave, Las Cruces, NM, 88003, USA

    Rebecca Creamer

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  1. Mohammad S. Alhawatema
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  4. Rebecca Creamer
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Correspondence to Rebecca Creamer.

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Alhawatema, M.S., Gebril, S., Cook, D. et al. RNAi-mediated down-regulation of a melanin polyketide synthase (pks1) gene in the fungus Slafractonia leguminicola . World J Microbiol Biotechnol 33, 179 (2017). https://doi.org/10.1007/s11274-017-2346-y

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  • DOI: https://doi.org/10.1007/s11274-017-2346-y

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