Benomyl-resistant Beauveria bassiana (Hypocreales: Clavicipitaceae) mutants induced by ion beams

Yuyun Fitriana1, S. Shinohara1, Katsuya Satoh2, Issay Narumi2,3, Tsutomu Saito1
1Faculty of Agriculture, Shizuoka University, Shizuoka, Japan
2Quantum Beam Science Directorate, Japan Atomic Energy Agency, Gunma, Japan
3Faculty of Life Sciences, Toyo University, Gunma, Japan

Tóm tắt

Abstract

Fungicides targeted at phytopathogens can be harmful to entomopathogenic fungi. The purpose of this study was to use ion-beam irradiation technology to produce benomyl-resistant mutants of the entomopathogenic fungus Beauveria bassiana (Bals.-Criv.) Vuill. After irradiation of conidia at 150 Gy, two mutant isolates, BB22 and BB24, were selected on media containing the fungicide. In an assay of vegetative growth, BB22 and BB24 were over 500 and 800 times more tolerant to benomyl, respectively, compared with the wild isolate. However, in an assay of conidial germination, neither mutant had increased tolerance compared with the wild isolate. Both mutant isolates also had increased tolerance to thiophanate-methyl during vegetative growth, but reduced tolerance to diethofencarb. A mutation was found at position 198 of the β-tubulin gene in the mutant isolates, with a substitution of glutamate for alanine (E198A). Ion beams have great potential as a tool to improve the traits of entomopathogenic fungi such as increasing tolerance to fungicides. Fungicide-resistant mutants produced in this way could be useful agents for biological control within IPM programmes where fungicides are also used.

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Tài liệu tham khảo

Abbott WS (1925) A method of computing the effectiveness of an insecticide. J Econ Entmol 18:265–267

Albertini C, Gredt M, Leroux P (1999) Mutations of the β-tubulin gene associated with different phenotypes of benzimidazole resistance in the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis. Pestic Biochem Physiol 64:17–31

Boucias DG, Pendland JC (1998) Principles of insect pathology. Kluwer Academic Publishers, London

Butters JA, Devi KU, Mohan CM, Sridevi V (2003) Screening for tolerance to bavistin, a benzimidazole fungicide containing methyl benzimidazol-2-yl carbamate (MBC), in Beauveria bassiana: sequence analysis of β-tubulin gene to identify mutations conferring tolerance. Mycol Res 107:260–266

Clark RA, Casagrande RA, Wallace DB (1982) Influence of pesticides on Beauveria bassiana, a pathogen of the Colorado potato beetle. Environ Entomol 11:67–70

Copping LG (2009) The manual of biological agents, 4th edn. BCPC, Hampshire

D’Alessandro CP, Padin S, Urrutia MI, López Lastra CC (2011) Interaction of fungicides with the entomopathogenic fungus Isaria fumosorosea. Biocontrol Sci Techn 21:189–197

Das SK, Masuda M, Hatashita M, Sakurai A, Sakakibara M (2008) A new approach for improving cordycepin productivity in surface liquid culture of Cordyceps militaris using high-energy ion beam irradiation. Lett Appl Microbiol 47:534–538

Davidse LC (1986) Benzimidazole fungicides: mechanism of action and biological impact. Annu Rev Phytopathol 24:43–65

Davidson RM, Hanson LE, Franc GD, Panella L (2006) Analysis of β-tubulin gene fragments from benzimidazole-sensitive and -tolerant Cercospora beticola. J Phytopathol 154:321–328

Fujimura M, Oeda K, Inoue H, Kato T (1992) A single amino-acid substitution in the beta-tubulin gene of Neurospora confers both carbendazim resistance and diethofencarb sensitivity. Curr Genet 21:399–404

Goettel MS, Inglis GD (1997) Fungi: Hyphomycetes. In: Lancey LA (ed) Manual of techniques in insect pathology. Academic Press, London, pp 213–249

Hollomon DW, Jenny A, Butters JA, Barker H, Hall L (1998) Fungal β-tubulin, expressed as a fusion protein, binds benzimidazole and phenylcarbamate fungicides. Antimicrob Agents Chemother 42:2171–2173

Inglis GD, Goettel MS, Butt TM, Strasser H (2001) Use of Hyphomycetous fungi for managing insect pests. In: Butt TM, Jackson C, Magan N (eds) Fungi as biological control agents: progress, problems and potential. CABI Publishing, Oxon, pp 23–69

James RR, Croft BA, Shaffer BT, Lighthart B (1998) Impact of temperature and humidity on host-pathogen interactions between Beauveria bassiana and a coccinellid. Biol Control 27:1506–1513

Keinath AP, Zitter TA (1998) Resistance to benomyl and thiophanate-methyl in Didymella bryoniae from South Carolina and New York. Plant Dis 82:479–484

Kim SK, Shim HJ, Roh JY, Jin BR, Boo KS, Je YH (2005) Isolation and characterization of benomyl-resistant mutants in an entomopathogenic fungus, Metarhizium anisopliae. Int J Indust Entomol 10:119–123

Koenraadt H, Somerville SC, Jones AL (1992) Characterization of mutations in the beta-tubulin gene of benomyl-resistant field strains of Venturia inaequalis and other plant pathogenic fungi. Phytopathology 82:1348–1354

Koo BS, Park H, Kalme S, Park HY, Han JW, Yeo YS, Yoon SH, Kim SJ, Lee CM, Yoon MY (2009) α- and β-tubulin from Phytophthora capsici KACC 40483: molecular cloning, biochemical characterization, and antimicrotubule screening. Appl Microbiol Biotechnol 82:513–524

Ma Z, Yoshimura MA, Michailides TJ (2003) Identification and characterization of benzimidazole resistance in Monilinia fructicola from stone fruit orchards in California. Appl Environ Microbiol 69:7145–7152

Matuo Y, Nishijima S, Hase Y, Sakamoto A, Tanaka A, Shimizu K (2006) Specificity of mutations induced by carbon ions in budding yeast Saccharomyces cerevisiae. Mutat Res 602:7–13

Qiu J, Xu J, Yu J, Bi C, Chen C, Zhou M (2011) Localisation of the benzimidazole fungicide binding site of Gibberella zeae β2-tubulin studied by site-directed mutagenesis. Pest Manag Sci 67:191–198

Saito T (1984) Effect of pesticides on conidial germination and hyphal growth of the entomopathogenic fungus Beauveria bassiana. Jpn J Appl Entomol Zool 28:87–89 (in Japanese with English summary)

Shinohara S, Fitriana Y, Satoh K, Narumi I, Saito T (2013) Enhanced fungicide resistance in Isaria fumosorosea following ionizing radiation-induced mutagenesis. FEMS Microbiol Lett 349:54–60

SPSS (2009) PASW statistics 18. SPSS Inc., Chicago

Tanaka A, Shikazono N, Hase Y (2010) Studies on biological effects of ion beams on lethality, molecular nature of mutation, mutation rate, and spectrum of mutation phenotype for mutation breeding in higher plants. J Radiat Res 51:223–233

Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

Toyoshima Y, Takahashi A, Tanaka H, Watanabe J, Mogi Y, Yamazaki T, Hamada R, Iwashita K, Satoh K, Narumi I (2012) Lethal and mutagenic effects of ion beams and γ-rays in Aspergillus oryzae. Mutat Res 740:43–49

Valadares-Inglis MC, Inglis PW (1997) Transformation of the entomopathogenic fungus, Metarhizium flavoviride strain CG423 to benomyl resistance. FEMS Microbiol Lett 155:199–202

Ziogas BN, Girgis SM (1993) Cross-resistance relationships between benzimidazole fungicides and diethofencarb in Botrytis cinerea and their genetical basis in Ustilago maydis. Pestic Sci 39:199–205

Zou G, Ying SH, Shen ZC, Feng MG (2006) Multi-sited mutations of β-tubulin are involved in benzimidazole resistance and thermotolerance of fungal biocontrol agent Beauveria bassiana. Environ Microbiol 8:2096–2105