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Metarhizium robertsii and M. acridum conidia produced on riboflavin-supplemented medium have increased UV-A tolerance and upregulated photoprotection and photoreactivation genes

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Abstract

The aim of this study was to evaluate the effect of riboflavin supplementation of culture medium on conidial UV-A tolerance of M. acridum (Driver & Milner) (Hypocreales: Clavicipitaceae) and M. robertsii (Bischoff, Rehner & Humber) (Hypocreales: Clavicipitaceae). These fungi were produced on culture medium supplemented, or not supplemented, with riboflavin. Relative germination and expression patterns of some photoprotection-related genes were evaluated after irradiating with artificial UV-A, or with filtered solar radiation (> 320 nm; UV-A and visible radiation). M. acridum conidia harvested from riboflavin-supplemented culture medium demonstrated enhanced UV-A tolerance when irradiated with artificial UV-A. Nevertheless, relative germination of conidia of both species produced on riboflavin-supplemented medium and exposed to filtered solar radiation was significantly higher than those produced on medium not supplemented with riboflavin. Riboflavin increased the transcription of photolyases, laccases and polyketide synthase genes. However, each fungal species induced different genes patterns involved in DNA repair and photoprotection. The addition of riboflavin to the substrate used for mass production of Metarhizium spp. and the resulting enhancement of conidial tolerance to solar radiation may improve the effectiveness of these fungi in biological control programs.

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Acknowledgements

We are grateful to Richard A. Humber (USDA/ARS, Ithaca, NY, USA) for providing the fungal isolates used in this study, and to Ricardo N. Marreto (Faculdade de Farmácia/UFG, Goiânia, GO, Brazil) for providing access to the Caron photostability chamber used for our artificial UV-A and white-light exposure tests. We also thank the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) of Brazil for providing PhD scholarship for R.A. Pereira-Junior, MSc scholarship for F.R.S. Paixão, and sandwich scholarships for both R.A. Pereira-Junior and F.R.S. Paixão (PPCP CAPES/Mercosul-016/2011). This research was supported by grants from the Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG, 201210267001060), CAPES (PPCP 016/2011), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) of Brazil (484329/2012-0), Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil (INCT 465678/2014-9), and Cooperative Agreements between D.W. Roberts and the United States Department of Agriculture (USDA – APHIS #14-8130-0114 and #14-8130-0368). We thank CNPq for grants PQ 308850/2015-7 to É.K.K. Fernandes, PQ 308189/2013-2 to C. Luz, and PQ 308505/2015-8 to G.U.L. Braga. We also thank the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) for grant #5055/14 to N. Pedrini. N. Pedrini is member of the CONICET Researcher’s Career, Argentina.

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Author notes

  1. Carla Huarte-Bonnet and Ronaldo A. Pereira-Junior have contributed equally to this work.

Authors and Affiliations

  1. Instituto de Patologia Tropical E Saúde Pública, Universidade Federal de Goiás, Avenida Esperança s/n, Campus Samambaia, Goiânia, GO, 74690-900, Brazil

    Carla Huarte-Bonnet, Ronaldo A. Pereira-Junior, Flávia R. A. Paixão, Christian Luz & Éverton K. K. Fernandes

  2. Instituto de Investigaciones Bioquímicas de La Plata – INIBIOLP, CONICET - Facultad de Ciencias Médicas, Universidad Nacional de La Plata, 1900, La Plata, Argentina

    Carla Huarte-Bonnet, Ronaldo A. Pereira-Junior, Flávia R. A. Paixão & Nicolás Pedrini

  3. Centro Universitário de Goiás – Uni-ANHANGUERA, Goiânia, GO, Brazil

    Ronaldo A. Pereira-Junior

  4. Departamento de Análises Clínicas, Toxicológicas E Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    Gilberto U. L. Braga

  5. Department of Biology, Utah State University, Logan, UT, USA

    Donald W. Roberts

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  1. Carla Huarte-Bonnet
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  2. Ronaldo A. Pereira-Junior
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Correspondence to Nicolás Pedrini or Éverton K. K. Fernandes.

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10526_2019_9990_MOESM1_ESM.pptx

Supplementary file1—Supplementary Fig. S1 Spectral irradiance of: (a) UV-A radiation emitted by the lamps in the indoor photostability chamber (Series 6540, Caron Products & Services Inc., Marietta, OH, USA), at its ‘UV-A’ setting with irradiance of 16,000 mW m-2; (b) Solar radiation filtered by a 0.1 mm-thick clear polyester film (JSC Industries, La Mirada, USA) to remove UV-B radiation, resulting in irradiance of 19,126 mW m-2 of solar UV-A radiation based on out-door measurements performed on three different sunny days in Goiânia, Goiás, Brazil (Center West Brazil: 16º40′30″ South and 49º14′42″ West, 749 m elevation), at noon from June 16th to July 9th, 2017; (c) White light emitted by the photostability chamber at its ‘white light’ setting (Series 6540, Caron Products & Services Inc., Marietta, OH, USA), at 16 Klux (PPTX 77 kb)

Supplementary file2—Supplementary Table S1 (DOC 45 kb)

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Huarte-Bonnet, C., Pereira-Junior, R.A., Paixão, F.R.A. et al. Metarhizium robertsii and M. acridum conidia produced on riboflavin-supplemented medium have increased UV-A tolerance and upregulated photoprotection and photoreactivation genes. BioControl 65, 211–222 (2020). https://doi.org/10.1007/s10526-019-09990-w

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