Abstract
Deep sequencing of small RNAs has proved effective in the diagnosis of mycovirus infections. In this study, the presence of mycoviruses in ten isolates of the phytopathogenic fungus Fusarium circinatum was investigated by high-throughput sequencing (HTS) of small RNAs. The contigs resulting from de novo assembly of the reads were aligned to viral genome sequences. The presence of each mycovirus detected in the isolates was confirmed by RT-PCR analysis with four previously described primer pairs and seven new pairs designed on the basis of sequencing data. The findings demonstrate the potential use of HTS for reconstructing previously identified mitoviruses infecting F. circinatum.
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Acknowledgements
The authors thank Instituto Agroforestal Mediterráneo (Universidad Politécnica de Valencia) for providing the isolates Va70, FC5, FC13, FC14, FC20 and FC24. The authors also thank Diana Bezos for performing the morphological and molecular identification of the isolates FC122, FC179, FC213 and FC921, Milagros de Vallejo and Juan Blanco (Gobierno de Cantabria) for their help in carrying out the study, and Elena Hidalgo (iuFOR, UVa-INIA), María Teresa Pérez-García, Pilar Cidad and Marycarmen Arévalo (IBGM, UVa-CSIC) for their collaboration and valuable advice during laboratory work.
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This study was supported by two research projects: AGL2015-69370-R “Next Generation Sequencing NGS) technologies for the study of Fusarium circinatum mycoviruses” (MINECO/FEDER, UE) and AGL2012-39912 “Biological control of Pine Pitch Canker disease by the use of Fusarium circinatum mycoviruses” (Ministerio Economía y Competitividad). This article is based upon work from COST Action FP1406 PINESTRENGTH (Pine pitch canker - strategies for management of Gibberella circinata in greenhouses and forests), supported by COST (European Cooperation in Science and Technology). E. J. Muñoz-Adalia is a receipt of grants from the European Social Fund and from the Consejería de Educación de Castilla y León (JCyL) (ORDEN EDU/1083/2013).
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Muñoz-Adalia, E.J., Diez, J.J., Fernández, M.M. et al. Characterization of small RNAs originating from mitoviruses infecting the conifer pathogen Fusarium circinatum. Arch Virol 163, 1009–1018 (2018). https://doi.org/10.1007/s00705-018-3712-2
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DOI: https://doi.org/10.1007/s00705-018-3712-2