The influence of tin ions on growth and enzymatic activity of entomopathogenic fungi

Łukasz Łopusiewicz, Kinga Mazurkiewicz-Zapałowicz, Marta Koniuszek, Cezary Tkaczuk, Artur Bartkowiak


In this in vitro study, the influence of tin ions at concentrations of 1–1,000 ppm on the development and enzymatic activity of four entomopathogenic fungi (Beauveria bassiana, B. brongniartii, Isaria fumosorosea, and Metarhizium robertsii), that are commonly used in biological plant protection, are examined. Each of the fungal species tested reacted differently to contact with the Sn2+ ions at the tested concentrations. Exposure to Sn2+ ions affected the rate of development, morphology, and enzymatic activity of fungi. Of the four fungal species studied, M. robertsii was the most resistant and showed complete growth inhibition at the highest Sn2+ concentration tested (1,000 ppm). For the other entomopathogenic fungi, the fungicidal effect of Sn2+ ions was noted at the concentration of 750 ppm. Exposure to Sn2+ ions (up to 500 ppm) resulted in enhanced biochemical activity; and all entomopathogens that were tested showed increased production of N-acetyl-β-glucosaminidase (NAG) as well as several proteases. Moreover, B. brongniartii and M. roberstii showed increased lipases synthesis. These changes may increase the pathogenicity of the fungi, thereby making them more effective in limiting the population of pest insects. The exposure of the entomopathogenic fungi to a medium containing Sn2+ ions, at concentrations that were appropriate for each species, induced hyperproduction of hydrolases, which might be involved in aiding the survival of entomopathogenic fungi in the presence of heavy metals. This study shows that the fungistatic effect of Sn2+ on entomopathogenic fungi did not restrict their pathogenicity, as evidenced by the stimulation of the production of enzymes that are involved in the infection of insects.


heavy metals; Beauveria; Isaria; Metarhizium; microfungi

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Tkaczuk C. The effect of selected heavy metals ions on the growth and conidial germination of the aphid pathogenic fungus Pandora neoaphidis (Remaudiére et Hennebert) Humber. Pol J Environ Stud. 2005;14:897–902.

Ropek D, Para A. The effect of heavy metal ions and their complexions upon the growth, sporulation and pathogenicity of the entomopathogenic fungus Verticillium lecanii. J Invertebr Pathol. 2002;79:123–125.

Tripathi P, Srivastava S. Mechanism to combat cobalt toxicity in cobalt resistant mutants of Aspergillus nidulans. Indian J Microbiol. 2007;47:336–344.

Ezzouhri L, Castro E, Moya M, Espinola F, Lairini K. Heavy metal tolerance of filamentous fungi isolated from polluted sites in Tangier, Morocco. Afr J Microbiol Res. 2009;3:35–48.

Singh D, Raina TK, Sing J. Entomopathogenic fungi: an effective biocontrol agent for management of insects populations naturally. Journal of Pharmaceutical Sciences and Research. 2017;9(6):830–839.

Pečiulytė D, Dirginčiutė-Volodkienė V. Effect of zinc and copper on cultivable populations of soil fungi with special reference to entomopathogenic fungi. Ekologija. 2012;58:65–85.

Ferron P. Pest control by the fungi Beauveria and Metarhizium. In: Burges HD, editor. Microbial control of pest and plant diseases. London: Academic Press; 1981. p. 465–481.

Keller S, Zimmermann G. Mycopathogens of soil insects. In: Wilding N, Collins NM, Hammond PM, Webber JF, editors. Insect–fungus interactions. London: Academic Press; 1989. p. 239–270. (Symposium of the Royal Entomological Society; vol 14).

Hajek AE. Ecology of terrestrial fungal entomopathogens. Adv Microb Ecol. 1997;15:193–249.

Baldrian P. Interactions of heavy metals with white-rot fungi. Enzyme Microb Technol. 2003;32:78–91.

Hassn WA, Asaf LH, Salih MSM. Effect of heavy metals ions on growth, sporulation and pathogenicity of Isaria javanica = (Paecilomyces javanicus). Int J Pure Appl Sci Technol. 2014;20(2):1–7.

Tkaczuk C, Majchrowska-Safaryan A, Panasiuk T, Tipping C. Effect of selected heavy metal ions on the growth of entomopathogenic fungi from the genus Isaria. Appl Ecol Environ Res. 2019;17(2):2571–2582.

Ashraf MA, Maah MJ, Yusoff I. Heavy metals accumulation in plants growing in ex tin mining catchment. International Journal of Environmental Science and Technology. 2011;8:401.

Müller F, Cyster L, Raitt L, Aalbers J. The effects of tin (Sn) additions on the growth of spinach plants. Phyton. 2016;84(2):461–465.

El-Makawy AI, Girgis SM, Khalil WK. Developmental and genetic toxicity of stannous chloride in mouse dams and fetuses. Mutat Res. 2008;657(2):105–110.

Tchounwou PB, Yedjou CG, Patlolla AK, Sutton DJ. Heavy metal toxicity and the environment. Exp Suppl. 2012;101:133–164.

Tobin JM, Cooney JJ. Action of inorganic tin and organotins on a hydrocarbon-using yeast, Candida maltosa. Arch Environ Contam Toxicol. 1999;36(1):7–12.

Lascourrèges J, Caumette P, Donard OF. Toxicity of butyltin, phenyltin and inorganic tin compounds to sulfate‐reducing bacteria isolated from anoxic marine sediments. Appl Organometal Chem. 2000;14:98–107.<98::AID-AOC962>3.0.CO;2-4

Mishra A, Mishra KP. Bacterial response as determinant of oxidative stress by heavy metals and antibiotic. Journal of Innovations in Pharmaceuticals and Biological Sciences. 2015;2(3):229–239.

Bernat P, Gajewska E, Szewczyk R, Słaba M, Długoński J. Tributyltin (TBT) induces oxidative stress and modifies lipid profile in the filamentous fungus Cunninghamella elegans. Environ Sci Pollut Res. 2014;21:4228.

Soboń A, Szewczyk R, Długoński J. Tributyltin (TBT) biodegradation induces oxidative stress of Cunninghamella echinulata. Int Biodeterior Biodegradation. 2016;107:92–101.

Şişman T. Early life stage and genetic toxicity of stannous chloride on zebrafish embryos and adults: toxic effects of tin on zebrafish. Environ Toxicol. 2011;26(3):240–249.

Sunday AO, Alafara BA, Olutona Godwin Oladele OG. Toxicity and speciation analysis of organotin compounds. Chemical Speciation and Bioavailability. 2012;24:216–226.

Cooney JJ, Wuertz S. Toxic effects of tin compounds on microorganisms. J Ind Microbiol. 1989;4:375–402.

Zimmermann G. The “Galleria bait method” for detection of entomopathogenic fungi in soil. J Appl Entomol. 1986;2:213–215.

Fazli M, Soleimani N, Mehrasbi M, Darabian S, Mohammadi J, Ramazani A. Highly cadmium tolerant fungi: their tolerance and removal potential. J Environ Health Sci Eng. 2015;13:19.

Hallas LE, Thayer JS, Cooney JJ. Factors affecting the toxic effect of tin on estuarine microorganisms. J Appl Environ Microbiol. 1982;44:193–197.

Kähkönen MA, Miettinen O, Kinnunen A, Hatakka A. Effects of gadolinium and tin to the production of oxidative enzymes and the growth of five basidiomycetous fungi. Expert Opin Environ Biol. 2017;6(1):1000139.

Kothandaraman K, Dawood Sharief S. Effect of Moringa oleifera against stannous chloride toxicity in rats (Rattus norvegicus). International Journal of Pharmaceutical and Biological Archives. 2013;4(4):771–774.

Dantas FJ, Moraes MO, de Mattos JC, Bezerra RJ, Carvalho EF, Filho MB, et al. Stannous chloride mediates single strand breaks in plasmid DNA through reactive oxygen species formation. Toxicol Lett. 1999;110(3):129–136.

Viau C, Pungartnik C, Schmitt MC, Basso TS, Henriques JA, Brendel M. Sensitivity to Sn2+ of the yeast Saccharomyces cerevisiae depends on general energy metabolism, metal transport, anti-oxidative defences, and DNA repair. Biometals. 2006;19(6):705–714.

Bernardo-Filho M, Cunha Mda C, Valsa Ide O, de Araujo AC, da Silva FC, da Fonseca Ade S. Evaluation of potential genotoxicity of stannous chloride: inactivation, filamentation and lysogenic induction of Escherichia coli. Food Chem Toxicol. 1994;32(5):477–479.

McLean JR, Blakey DH, Douglas GR, Kaplan JG. The effect of stannous and stannic (tin) chloride on DNA in Chinese hamster ovary cells. Mutat Res. 1983;119(2):195–201.

Anahid S, Yaghmaei S, Ghobadinejad Z. Heavy metal tolerance of fungi. Scientia Iranica. 2011;18:502–508.

Rasha FM. Intracellular siderophore detection in an Egyptian, cobalt-treated F. solani isolate using SEM-EDX with reference to its tolerance. Pol J Microbiol. 2017;66(2):235–243.

Colpaert J, Vandenkoornhuyse P, Adriaensen K, Vangronsveld J. Genetic variation and heavy metal tolerance in the ectomycorrhizal basidiomycete Suillus luteus. New Phytol. 2000;147(2):367–379.

Hu Q, Li F, Zhang Y. Risks of mycotoxins from mycoinsecticides to humans. Biomed Res Int. 2016;2016:3194321.

Pusztahelyi T, Pócsi I. Chitinase but N-acetyl-β-d-glucosaminidase production correlates to the biomass decline in Penicillium and Aspergillus species. Acta Microbiol Immunol Hung. 2014;61:131–143.

Beys da Silva WO, Santi L, Schrank A, Vainstein MH. Metarhizium anisopliae lipolytic activity plays a pivotal role in Rhipicephalus (Boophilus) microplus infection. Fungal Biol. 2010;114(1):10–15.

Pedrini N, Ortiz-Urquiza A, Huarte-Bonnet C, Zhang S, Keyhani NO. Targeting of insect epicuticular lipids by the entomopathogenic fungus Beauveria bassiana: hydrocarbon oxidation within the context of a host–pathogen interaction. Front Microbiol. 2013;4:24.

Wang C, Typas MA, Butt TM. Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae. FEMS Microbiol Lett. 2002;213:251–255.

Sánchez-Pérez L, Barranco-Florido J, Rodríguez-Navarro S, Cervantes-Mayagoitia J, Ramos-López M. Enzymes of entomopathogenic fungi, advances and insights. Adv Enzyme Res. 2014;2:65–76.