Abstract
Chlorate resistant mutants, which were first isolated in the zygomycetous fungusPhycomyces blakesleeanus, were found to be resistant up to a concentration of at least 300 mM of potassium chlorate. The dose-response relationship showed that although the mutants could be divided into two groups based on chlorate resistance in the mycelial elongation assay on the solid minimal medium, this was not observed in the assay using liquid culture. Genetic analysis of heterokaryons revealed the mutant alleles to be dominant. Enzymatic activities of three nitrate reductases and chlorate reductase were deficient in both the parent strain and the mutants. Intracellular incorporation of chlorate ion varied from strain to strain; however, the variation could not explain the mechanism of chlorate resistance. One unexpected characteristic of the mutants was that the intracellular sulfate ion concentration was 3.5 to 5.5 times higher than in the parent strain. We designated this mutant genotypecrw, chlorate resistant mutant from nitrate-nonutilizing wild type.
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Literature cited
Alvarez, M. I. and Eslava, A. P. 1983. Isogenic strains ofPhycomyces blakesleeanus suitable for genetic analysis. Genetics105: 873–879.
Alvarez, M. I., Peláez, M. I. and Eslava, A. P. 1980. Recombination between ten markers inPhycomyces. Mol. Gen. Genet.179: 447–452.
Brownlee, A. G. and Arst Jr., H. N. 1983. Nitrate uptake inAspergillus nidulans and the involvement of the third gene of the nitrate assimilatory gene cluster. J. Bacteriol.115: 1138–1146.
Cerdá-Olmedo, E. 1987. Standard growth conditions and variations. In:Phycomyces, (ed. by Cerdá-Olmedo, E. and Lipson, E. D.), pp. 337–339. Cold Spring Harbor Laboratory, New York.
Cove, D. J. 1976. Chlorate toxicity inAspergillus nidulans. Studies of mutants altered in nitrate assimilation. Mol. Gen. Genet.146: 147–159.
Davies, J. and Smith, D. I. 1978. Plasmid-determined resistance to antimicrobial agents. Ann. Rev. Microbiol.32: 469–518.
Davis, B. D. and Maas, W. K. 1952. Analysis of the biochemical mechanism of drug resistance in certain bacterial mutants. Proc. Natl. Acad. Sci. U.S.A.38: 775–785.
Delbrück, M. and Ootaki, T. 1979. An unstable nuclear gene inPhycomyces. Genetics92: 27–48.
Foster, T. J. 1983. Plasmid-determined resistance to antimicrobial drugs and toxic metal ions in bacteria. Microbiol. Rev.47: 361–409.
Garcés, R., Santero, E. and Medina, J. R. 1984. Mutantes dePhycomyces blakesleeanus resistentes al alcohol alflico. Genet. Iber.36: 133–145.
Garcés, R., Pollock, J. A. and Lipson, E. D. 1985. Examination ofPhycomyces blakesleeanus for nitrate reductase as a possible blue light photoreceptor. Plant Sci.40: 173–177.
Garrett, R. H. and Cove, D. J. 1976. Formation of NADPH-nitrate reductase activity in vitro fromAspergillus nidulans niaD andcnx mutants. Mol. Gen. Genet.149: 179–186.
Garrett, R. H. and Nason, A. 1969. Further purification and properties ofNeurospora nitrate reductase. J. Biol. Chem.244: 2870–2882.
Griffin, D. H. 1994. Fungal physiology, 2nd ed., pp. 158–194. Wiley-Liss, New York.
Heisengerg, M. and Cerdá-Olmedo, E. 1968. Segregation of heterokaryons in the asexual cycle ofPhycomyces. Mol. Gen. Genet.102: 187–195.
Hilgenberg, W., Burke, P. V. and Sandmann, G. 1987. Metabolic pathways. In:Phycomyces, (ed. by Cerdá-Olmedo, E. and Lipson, E. D.), pp. 155–198. Cold Spring Harbor Laboratory, New York.
Lanfaloni, L., Cappanna, E. and Gualerzi, C. O. 1994. Isolation and characterization of a chlorate resistant mutant ofSpirulina platensis. Microbiologia17: 133–140.
Lewis, C. M. and Fincham, J. R. S. 1970. Genetics of nitrate reductase inUstilago maydis. Genet. Res. Camb.16: 151–163.
Liu, W. and Sundheim, L. 1996. Nitrate nonutilizing mutants and vegetative compatibility groups inFusarium poae. Fungal Genet. Biol.20: 12–17.
Medina, J. R. 1977. Continuous variation of genic dosage inPhycomyces. Genet. Res.30: 211–219.
Meissner, G. and Delbrück, M. 1968. Carotenes and retinal inPhycomyces mutants. Plant Physiol.43: 1279–1283.
Moussa, C., Houziaux, P., Danree, B. and Azerad, R. 1997. Fungal metabolism of phenolic and nonphenolicp-cymene-related drugs and prodrugs: I. Metabolites of thymoxamine. Drug Metab. Disposition25: 301–310.
Nicholas, D. J. D. and Nason, A. 1957. Determination of nitrate and nitrite. In: Methods in enzymology, vol. 3, (ed. by Colowick, S. P. and Kaplan, N. O.), pp. 981–984.
Ootaki, T. 1987. Heterokaryon formation. In:Phycomyces, (ed. by Cerdá-Olmedo, E. and Lipson, E. D.), pp. 346–349. Cold Spring Harbor Laboratory, New York.
Pichinoty, F. J., Puig, J., Chippaux, M. and Bigliardi-Rouvier, J. et Gendre, J. 1969. Recherches sur des mutants bactériens ayant perdu les activités catalytiques liées a la nitrate-réductase A. II. Comportement envers le chlorate et le chlorite. Ann. L'institut Pasteur116: 409–432.
Pothuluri, J. V., Evans, F. E., Heinze, T. M. and Cerniglia, C. E. 1996. Formation of sulfate and glucoside conjugates of benzo[e]pyrene byCunninghamella elegans. Appl. Microbiol. Biotech.45: 677–683.
Pothuluri, J. V., Sutherland, J. B., Freeman, J. P. and Cerniglia C. 1998. Fungal biotransformation of 6-nitrochrysene. Appl. Enviorn. Microbiol.64: 3106–3109.
Ryan, F. J., Beedle, G. W. and Tatum, E. L. 1943. The tube method of measuring the growth rate ofNeurospora. Am. J. Bot.30: 784–799.
Sasaki, H., Kataoka, H., Kamiya, M. and Kawai, H. 1999. Accumulation of sulfuric acid in Dictyotales (Phaeophyceae): Taxonomic distribution and ion chromatography of cell extracts. J. Phycol.35: 732–739.
Sutter, R. P. 1975. Mutations affecting sexual development inPhycomyces blakesleeanus. Proc. Natl. Acad. Sci. U.S.A.72: 127–130.
Unkles, S. E., Campbell, E. I., de Ruiter-Jacobs, Y. M. J. T., Broekhuijsen, M., Marco, J. A., Carrez, D., Contreras, R., van del Hondel, C. A. M. J. J. and Kinghorn, J. R. 1989. The development of a homologous transformation system forAspergillus oryzae based on the nitrate assimilation pathway: A convenient and general selection system for filamentous fungal transformation. Mol. Gen. Genet.218: 99–104.
Vaillancorut, L. J. and Hanau, R. M. 1994. Nitrate-nonutilizing mutants used to study heterokaryosis and vegetative compatibility inGlomerella graminicola (Colletotrichum greaminicola). Exp. Mycol.18: 311–319.
Van Laere, A. J., Carlier, A. R. and Van Assche, J. A. 1976. Effect of 5-fluorouracil and cycloheximide on the early development ofPhycomyces blakesleeanus spores and the activity of N-acetyl-glucosamine synthesizing enzymes. Arch. Microbiol.108: 113–116.
Whitaker, A. 1976. Amino acid transport into fungi: An essay. Trans. Br. Mycol. Soc.67: 365–376.
Yamazaki, Y., Kataoka, H., Miyazaki, A., Watanabe, M. and Ootaki, T. 1996. Action spectra for photoinhibition of sexual development inPhycomyces blakesleeanus. Photochem. Photobiol.64: 387–392.
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Fukui, J., Miyazaki, A. & Ootaki, T. Isolation and characterization of chlorate resistant mutants from nitrate-nonutilizing fungusPhycomyces blakesleeanus . Mycoscience 41, 633–640 (2000). https://doi.org/10.1007/BF02460930
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DOI: https://doi.org/10.1007/BF02460930