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The presence of mycoviral infection attenuates the growth and pathogenicity of the phytopathogenic fungus Botrytis cinerea collected from strawberry fields in Pakistan

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Abstract

Botrytis cinerea is a phytopathogenic fungus that belongs to the family Sclerotiniaceae in the Ascomycota group. This pathogen infects economically important crops and is responsible for substantial preharvest and postharvest losses each year worldwide. Mycoviruses are commonly associated with Botrytis cinerea isolates. In this work, we report the hypovirulence potential of an endornavirus harboured by the Kst31c isolate of Botrytis cinerea collected from a strawberry field in Pakistan. Our analysis revealed the presence of a positive-sense single-stranded RNA (+ ssRNA) mono-segmented virus in this isolate. Next-generation sequencing of total RNA harvested from Kst31c revealed the presence of an endornavirus with a genome length of 13,765 nt comprising a single open reading frame (ORF) encoding a polyprotein of 4,546 amino acids. The identified polyprotein includes four conserved domains: a viral methyltransferase, an RNA-dependent RNA polymerase 2, and the two putative viral helicases (DExDc-like and Hel-1). No cysteine-rich regions were identified in this virus. Phylogenetic analysis placed it in a clade within the genus Betaendornavirus. The current virus was tentatively named Botrytis cinerea endornavirus 4 (BcEV4). A comparison of virus-free and virus-infected isogenic lines showed that BcEV4 was associated with hypovirulence, slow growth, and altered phenotype. Furthermore, these results were also confirmed by pathogenicity testing via an apple assay. The virus can be tested as a potential biocontrol agent against Botrytis cinerea infections.

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Data Availability

The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the Manuscript/Supplementary Material.

References

  • Bhatti, M. F., Jamal, A., Bignell, E. M., Petrou, M. A., & Coutts, R. H. (2012). Incidence of dsRNA mycoviruses in a collection of Aspergillus fumigatus isolates. Mycopathologia, 174, 323–326.

    Article  CAS  PubMed  Google Scholar 

  • Castillo, A., Cottet, L., Castro, M., & Sepúlveda, F. (2011). Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae. Virology Journal, 8(1), 1–7.

  • Dawe, A. L., & Nuss, D. L. (2013). Hypovirus molecular biology: From Koch’s postulates to host self-recognition genes that restrict virus transmission. Advances in Virus Research, 86, 109–147.

    Article  CAS  PubMed  Google Scholar 

  • Fukuhara, T. (2019). Endornaviruses: Persistent dsRNA viruses with symbiotic properties in diverse eukaryotes. Virus Genes, 55, 165–173.

    Article  CAS  PubMed  Google Scholar 

  • Garcia-Pedrajas, M. D., Canizares, M. C., Sarmiento-Villamil, J. L., Jacquat, A. G., & Dambolena, J. S. (2019). Mycoviruses in Biological Control: From Basic Research to Field Implementation. Phytopathology, 109, 1828–1839.

    Article  CAS  PubMed  Google Scholar 

  • Ghabrial, S. A., Caston, J. R., Jiang, D., Nibert, M. L., & Suzuki, N. (2015). 50-plus years of fungal viruses. Virology, 479–480, 356–368.

    Article  PubMed  Google Scholar 

  • Hao, F., Zhou, Z., Wu, M., & Li, G. (2017). Molecular characterization of a novel endornavirus from the phytopathogenic fungus Botrytis cinerea. Archives of Virology, 162(1), 313–316.

    Article  CAS  PubMed  Google Scholar 

  • Jiang, Y., Wang, J., Yang, B., Wang, Q., Zhou, J., & Yu, W. (2019). Molecular Characterization of a Debilitation-Associated Partitivirus Infecting the Pathogenic Fungus Aspergillus flavus. Frontiers in Microbiology, 10, 626.

    Article  PubMed  PubMed Central  Google Scholar 

  • Khan, H. A., Sato, Y., Kondo, H., Jamal, A., Bhatti, M. F., & Suzuki, N. (2021). A second capsidless hadakavirus strain with 10 positive-sense single-stranded RNA genomic segments from Fusarium nygamai. Archives of Virology, 166(262), 2711–2722.

    Article  CAS  PubMed  Google Scholar 

  • Kondo, H., Botella, L., & Suzuki, N. (2022). Mycovirus Diversity and Evolution Revealed/Inferred from Recent Studies. Annual Review of Phytopathology, 60.

  • Linder-Basso, D., Dynek, J. N., & Hillman, B. I. (2005). Genome analysis of Cryphonectria hypovirus 4, the most common hypovirus species in North America. Virology, 337(1), 192–203.

    Article  CAS  PubMed  Google Scholar 

  • Luo, X., Jiang, D., Xie, J., Jia, J., Duan, J., Cheng, J., & Lin, Y. (2022). Genome Characterization and Phylogenetic Analysis of a Novel Endornavirus That Infects Fungal Pathogen Sclerotinia sclerotiorum. Viruses, 14(3), 456.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nuss, D. L. (2005). Hypovirulence: Mycoviruses at the fungal–plant interface. J Nature Reviews Microbiology, 3, 632–642.

  • Nuss, D. L., & Koltin, Y. (1990). Significance of dsRNA Genetic Elements in Plant Pathogenic Fungi. Annual Review of Phytopathology, 28, 37–58.

    Article  CAS  PubMed  Google Scholar 

  • Okada, R., Alcala-Briseno, R. I., Escalante, C., Sabanadzovic, S., & Valverde, R. A. (2018). Genomic sequence of a novel endornavirus from Phaseolus vulgaris and occurrence in mixed infections with two other endornaviruses. Virus Research, 257, 63–67.

    Article  CAS  PubMed  Google Scholar 

  • Okada, R., Moriyama, H., Ogawara, T., Cizek, H., & Valverde, R. A. (2021). Complete nucleotide sequence of an alphaendornavirus isolated from common buckwheat (Fagopyrum esculentum). Archives of Virology, 166, 3483–3486.

    Article  CAS  PubMed  Google Scholar 

  • Osaki, H., Nakamura, H., Sasaki, A., Matsumoto, N., & Yoshida, K. (2006). An endornavirus from a hypovirulent strain of the violet root rot fungus, Helicobasidium mompa. Virus Research, 118, 143–149.

    Article  CAS  PubMed  Google Scholar 

  • Otulak-Kozieł, K., Kozieł, E., Escalante, C., & Valverde, R. A. (2020). Ultrastructural analysis of cells from bell pepper (Capsicum annuum) infected with bell pepper endornavirus. Frontiers in Plant Science, 11, 491.

  • Pearson, M. N., & Bailey, A. M. (2013). Viruses of botrytis. Advances in Virus Research, 86, 249–272.

    Article  PubMed  Google Scholar 

  • Pearson, M. N., Beever, R. E., Boine, B., & Arthur, K. (2009). Mycoviruses of filamentous fungi and their relevance to plant pathology. Molecular Plant Pathology, 10, 115–128.

    Article  CAS  PubMed  Google Scholar 

  • Picarelli, M., Forgia, M., Rivas, E. B., et al. (2019). Extreme Diversity of Mycoviruses Present in Isolates of Rhizoctonia solani AG2-2 LP From Zoysia japonica From Brazil. Frontiers in Cellular and Infection Microbiology, 9, 244.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Potgieter, C. A., Castillo, A., Castro, M., Cottet, L., & Morales, A. (2013). A wild-type Botrytis cinerea strain co-infected by double-stranded RNA mycoviruses presents hypovirulence-associated traits. Virology Journal, 10, 220.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ruiz-Padilla, A., Rodriguez-Romero, J., Gomez-Cid, I., Pacifico, D., & Ayllon, M. A. (2021). Novel Mycoviruses Discovered in the Mycovirome of a Necrotrophic Fungus. mBio, 12, e03705-20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Schumann, U., Smith, N. A., & Wang, M. B. (2013). A fast and efficient method for preparation of high-quality RNA from fungal mycelia. BMC Research Notes, 6, 71.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shamsi, W., Sato, Y., Jamal, A., et al. (2019). Molecular and biological characterization of a novel botybirnavirus identified from a Pakistani isolate of Alternaria alternata. Virus Research, 263, 119–128.

    Article  CAS  PubMed  Google Scholar 

  • Song, H. Y., Kim, N., Kim, D. H., & Kim, J. M. (2020). The PoV mycovirus affects extracellular enzyme expression and fruiting body yield in the oyster mushroom. Pleurotus Ostreatus. Sci Rep, 10, 1094.

    Article  CAS  PubMed  Google Scholar 

  • Tuomivirta, T. T., Kaitera, J., & Hantula, J. (2009). A novel putative virus of Gremmeniella abietina type B (Ascomycota: Helotiaceae) has a composite genome with endornavirus affinities. Journal of General Virology, 90(9), 2299–2305.

    Article  CAS  PubMed  Google Scholar 

  • Uchida, K., Sakuta, K., Ito, A., et al. (2021). Two Novel Endornaviruses Co-infecting a Phytophthora Pathogen of Asparagus officinalis Modulate the Developmental Stages and Fungicide Sensitivities of the Host Oomycete. Frontiers in Microbiology, 12, 633502.

    Article  PubMed  PubMed Central  Google Scholar 

  • Valverde, R. A., Khalifa, M. E., Okada, R., Fukuhara, T., Sabanadzovic, S., ICTV Report Consortium. (2019). ICTV virus taxonomy profile: Endornaviridae. Journal of General Virology, 100(8), 1204–1205.

    Article  CAS  PubMed  Google Scholar 

  • Wagemans, J., Holtappels, D., Vainio, E., Rabiey, M., Marzachì, C., Herrero, S., ... & Turina, M. (2022). Going viral: virus-based biological control agents for plant protection. Annual Review of Phytopathology, 60.

  • White, T. J., Bruns, T., Lee, S. J. W. T., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. PCR Protocols: A Guide to Methods and Applications, 18(1), 315–322.

    Google Scholar 

  • Williamson, B., Tudzynski, B., Tudzynski, P., & Van Kan, J. A. (2007). Botrytis cinerea: The cause of grey mould disease. Molecular Plant Pathology, 8, 561–580.

    Article  CAS  PubMed  Google Scholar 

  • Wu, M., Zhang, L., Li, G., Jiang, D., & Ghabrial, S. A. (2010). Genome characterization of a debilitation-associated mitovirus infecting the phytopathogenic fungus Botrytis cinerea. Virology, 406, 117–126.

    Article  CAS  PubMed  Google Scholar 

  • Yang, D., Wu, M., Zhang, J., Chen, W., Li, G., & Yang, L. (2018). Sclerotinia minor Endornavirus 1, a Novel Pathogenicity Debilitation-Associated Mycovirus with a Wide Spectrum of Horizontal Transmissibility. Viruses, 10, 589.

    Article  PubMed  PubMed Central  Google Scholar 

  • Yu, L., Sang, W., Wu, M. D., et al. (2015). Novel hypovirulence-associated RNA mycovirus in the plant-pathogenic fungus Botrytis cinerea: Molecular and biological characterization. Applied and Environment Microbiology, 81(305), 2299–2310.

    Article  CAS  Google Scholar 

  • Zheng, L., Shu, C., Zhang, M., Yang, M., & Zhou, E. (2019). Molecular Characterization of a Novel Endornavirus Conferring Hypovirulence in Rice Sheath Blight Fungus Rhizoctonia solani AG-1 IA Strain GD-2. Viruses, 11, 178.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhong, J., Chen, D., Zhu, H. J., Gao, B. D., & Zhou, Q. (2016). Hypovirulence of Sclerotium rolfsii caused by associated RNA mycovirus. Frontiers in Microbiology, 7, 1798.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

AA is thankful to the Higher Education Commission (HEC) of Pakistan for providing a fellowship under the International Research Support Initiative Program (IRSIP) and NUST and NARC for research facilitation.

Funding

The authors are grateful to NUST for providing Student research funds and Higher Education Commission (HEC) of Pakistan for providing research grant (NRPU Research Grant No. 20–4109/R&D/HEC/14/847) to MFB (PI) and NV (Co-PI) to conduct this investigation.

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Author notes

  1. Nasar Virk

    Present address: EBS Business School, EBS Universität für Wirtschaft und Recht, Rheingaustrasse 1, Oestrich-Winkel, 65375, Germany

Authors and Affiliations

  1. Atta-ur-Rahman School of Applied Biosciences (ASAB), National University of Sciences and Technology (NUST), Sector H-12, Islamabad, 44000, Pakistan

    Aqeel Ahmed, Haris Ahmed Khan, Danish Ilyas Baig, Tehsin Fatma, Nasar Virk & Muhammad Faraz Bhatti

  2. Department of Biotechnology, University of Mianwali, Mianwali, Pakistan

    Haris Ahmed Khan

  3. Crop Diseases Research Institute, National Agricultural Research Centre, Islamabad, Pakistan

    Atif Jamal

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  1. Aqeel Ahmed
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Contributions

AA: Screening. AA and HAK: Investigation, Writing—original draft. DB and TF: Investigation. AJ and NV: Reviewing & editing. MFB: Conceptualization, Supervision, Writing – reviewing & editing- original draft.

Corresponding author

Correspondence to Muhammad Faraz Bhatti.

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Ahmed, A., Khan, H.A., Jamal, A. et al. The presence of mycoviral infection attenuates the growth and pathogenicity of the phytopathogenic fungus Botrytis cinerea collected from strawberry fields in Pakistan. Eur J Plant Pathol 167, 133–143 (2023). https://doi.org/10.1007/s10658-023-02689-w

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