Evaluation of metal toxicity by a modified method based on the fungus Gerronema viridilucens bioluminescence in agar medium.
Metal cation toxicity to basidiomycete fungi is poorly understood, despite its well-known importance in terrestrial ecosystems. Moreover, there is no reported methodology for the routine evaluation of metal toxicity to basidiomycetes. In the present study, we describe the development of a procedure to assess the acute toxicity of metal cations (Na(+), K(+), Li(+), Ca(2+),Mg(2+), Co(2+), Zn(2+), Ni(2+), Mn(2+), Cd(2+), and Cu(2+)) to the bioluminescent basidiomycete fungus Gerronema viridilucens. The method is based on the decrease in the intensity of bioluminescence resulting from injuries sustained by the fungus mycelium exposed to either essential or nonessential metal toxicants. The assay described herein enables us to propose a metal toxicity series to Gerronema viridilucens based on data obtained from the bioluminescence intensity (median effective concentration [EC50] values) versus metal concentration: Cd(2+) > Cu(2+) > Mn(2+) approximately Ni(2+) approximately Co(2+) > Zn(2+) > Mg(2+) > Li(+) > K(+) approximately Na(+) > Ca(2+), and to shed some light on the mechanism of toxic action of metal cations to basidiomycete fungi.
Mendes
Luiz Fernando
LF
Instituto de Química da Universidade de São Paulo, Caixa Postal 26077, 05599-970 São Paulo, SP, Brazil.
Stevani
Cassius Vinicius
CV
Instituto de Química da Universidade de São Paulo, Caixa Postal 26077, 05599-970 São Paulo, SP, Brazil.
D000362:Agar; D001487:Basidiomycota / Q000187:drug effects* / Q000254:growth & development; D008163:Luminescent Measurements / Q000379:methods*; D008670:Metals / Q000633:toxicity*
D008670:Metals; D000362:Agar
Walker CH, Hopkin SP, Sibly RM, Peakall DB. 1996. Principles of Ecotoxicology. Taylor & Francis, London, UK.
Steinberg SM, Poziomek EJ, Engelmann WH, Rogers KR. 1995. A review of environmental applications of bioluminescence measurements. Chemosphere 30: 2155-2197.
Gianfreda L, Rao MA. 2008. Interactions between xenobiotics and microbial and enzymatic soil activity. Crit Rev Environ Sci Technol 38: 269-310.
Sochová I, Hofman J, Holoubek I. 2006. Using nematodes in soil ecotoxicology. Environ Int 32: 374-383.
Hobbelen PHF, Koolhaas JE, van Gestel CAM. 2006. Bioaccumulation of heavy metals in the earthworms Lumbricus rubellus and Aporrectodea caliginosa in relation to total and available metal concentrations in field soils. Environ Pollut 144: 639-646.
Landis WG, Yu M-H. 1995. Introduction to Environmental Toxicology. Lewis, Boca Raton, FL, USA.
Gadd GM. 2001. Fungi in Bioremediation. Cambridge University Press, Cambridge, UK.
Gadd GM. 1982. The use of solid medium to study effects of cadmium copper and zinc on yeast-like fungi: applicability and limitations. J Appl Bacteriol 50: 57-62.
Høiland K, Dybdahl HG. 1993. A micro-well method for estimating fungal response to metal ions. Response to aluminium by some saprophytic basidiomycete. Nord J Bot 13: 691-696.
Blaudez D, Jacob C, Turnau K, Colpaert JV, Ahonen-Jonnarth U, Finlay R, Botton B, Chalot M. 2000. Differential responses of ectomycorrhizal fungi to heavy metals in vitro. Mycol Res 140: 1366-1371.
Colpaert JV, Vandenkoornhuyse P, Adriaensen K, Vangronsveld J. 2000. Genetic variation and heavy metal tolerance in the ectomycorrhizal basidiomycete Suillus luteus. New Phytol 147: 367-379.
Hartley J, Cairney JWG, Meharg AA. 1997. Do ectomycorrhizal fungi exhibit adaptive tolerance to potentially toxic metals in the environment? Plant Soil 189: 303-319.
Weitz HJ, Campbell CD, Killham K. 2002. Development of a novel, bioluminescence-based, fungal biossay for toxicity testing. Environ Microbiol 4: 422-429.
Airth RL, Foerster GE. 1962. The isolation of catalytic components required for cell-free fungal bioluminescence. Arch Biochem Biophys 97: 567-573.
Shimomura O. 1989. Chemiluminescence of panal (a sesquiterpene) isolated from the luminous fungus Panellus stipticus. Photochem Photobiol 49: 355-360.
Desjardin DE, Oliveira AG, Stevani CV. 2008. Fungi bioluminescence revisited. Photochem Photobiol Sci 7: 170-182.
Bechara EJH, Costa C, Colepicolo P, Viviani V, Barros MP, Timmins GS, Lall AB, Terra WR, Ferreira C, Stevani CV, Torres MA. 2007. Chemical, biological and evolutionary aspects of beetle bioluminescence. Arch Organic Chem 8: 311-323.
Desjardin DE, Capelari M, Stevani CV. 2005. A new bioluminescent Agaric from São Paulo, Brazil. Fungal Divers 18: 9-14.
Mendes LF, Bastos EL, Desjardin DE, Stevani CV. 2008. Influence of culture conditions on mycelial growth and bioluminescence of Gerronema viridilucens. FEMS Microbiol Lett 282: 132-139.
McCloskey JT, Newman MC, Clark SB. 1996. Predicting the relative toxicity of metal ions using ion characteristics: Microtox® bioluminescence assay. Environ Toxicol Chem 15: 1730-1737.
Lofts S, Spurgeon DJ, Svendsen C, Tipping E. 2004. Deriving soil critical limits for Cu, Zn, Cd, and Pb: A method based on free ion concentrations. Environ Sci Technol 38: 3623-3631.
Timbrell J. 2000. Principles of Biochemical Toxicology. Taylor & Francis, London, UK.
Hill AV. 1913. The combinations of haemoglobin with oxygen and with carbon monoxide. Biochem J 7: 471-480.
Weiss JN. 1997. The Hill equation revisited: uses and misuses. FASEB J 11: 835-841.
Høiland K. 1995. Reaction of some decomposer basidiomycete to toxic elements. Nord J Bot 15: 305-318.
Colpaert JV, Van Assche JA. 1987. Heavy metal tolerance in some ectomycorrhizal fungi. Funct Ecol 1: 415-421.
Ross IS. 1975. Some effects of heavy metals on fungal cells. Trans BR Mycol Soc 64: 175-193.
Dineley KE, Votyakova TV, Reynolds IJ. 2003. Zinc inhibition of cellular energy production: Implications for mitochondria and neurodegeneration. J Neurochem 85: 563-570.
Gadd GM. 1993. Interactions of fungi with toxic metals. New Phytol 124: 25-60.
Alloway BJ, 1995. ed. Heavy Metals in Soils, 2nd ed. Springer, London, UK.
Dudka S, Adriano DC. 1997. Environmental impacts of metal ore mining and processing: A review. J Environ Qual 26: 590-602.
Helmisaari HS, Jerome J, Fritze H, Nieminen T, Palmgren P, Salemaa M, Vanha-Majamaa I. 1995. Copper in scots pine forests around a heavy metal smelter in southwestern Finland. Water Air Soil Pollut 85: 1727-1732.
Roszyk E, Szerszen L. 1988. Accumulation of heavy metals in arable layer of soils in the protection zones of copper smelters. Roczniki Gleboznawcze 39: 135-156.
Ray P, Tiwari R, Reddy UG, Adholeya A. 2005. Detecting the heavy metal tolerance level in ectomycorrhizal fungi in vitro. World J Microbiol Biotechnol 21: 309-315.
del Val C, Barea JM, Azcàn-Aguilar C. 1999. Assessing the tolerance to heavy metals of arbuscular mycorrhizal fungi isolated from sewage sludge-contaminated soils. Appl Soil Ecol 11: 261-269.
Atimanav Gaur A, Adholeya A. 2004. Prospects of arbuscular mycorrhizal fungi in phytoremediation of heavy metal contaminated soils. Curr Sci 86: 528-534.
Campbell CD, Hird M, Lumsdon DG, Meeussen JCL. 2000. The effect of EDTA and fulvic acid on Cd, Zn, and Cu toxicity to a bioluminescent construct (pUCD607) of Escherichia coli. Chemosphere 40: 319-325.
Sarin C, Hall JM, Cotter-Howells J, Killham K, Cresser MS. 2000. Influence of complexation with chloride on the responses of a lux-marked bacteria bioassay to cadmium, copper, lead, and mercury. Environ Toxicol Chem 19: 259-264.
Ishaque AB, Johnson L, Gerald T, Boucaud D, Okoh J, Tchounwou PB. 2006. Assessment of individual and combined toxicities of four non-essential metals (As, Cd, Hg, and Pb) in the microtox assay. Int J Environ Res Public Health 3: 118-120.
McCreight JD, Schroeder DB. 1982. Inhibition of growth of nine ectomycorrhizal fungi by cadmium, lead and nickel in vitro. Environ Exp Bot 22: 1-7.
Tam PCF. 1995. Heavy metal tolerance by ectomycorrhizal fungi and metal amelioration by Pisolithus tinctorius. Mycorrhiza 5: 181-187.
Colpaert JV, Van AJA. 1992. The effects of cadmium and the cadmium-zinc interaction on the axenic growth of ectomycorrhizal fungi. Plant Soil 145: 237-243.
Jones D, Muehlchen A. 1994. Effects of the potentially toxic metals, aluminum, Zn, and Cu, on ectomycorrhizal fungi. J Environ Sci Health Part A 29: 949-966.
Willenborg A, Schmitz D, Lelley J. 1990. Effects of environmental stress factors on ectomycorrhizal fungi in vitro. Can J Bot 68: 1741-1746.
Guillén Y, Machuca A. 2008. The effect of copper on the growth of wood-rotting fungi and a blue-stain fungus. World J Microbiol Biotechnol 24: 31-37.