Journal of Genetics

, 77:71 | Cite as

Cloning of the sterol C-14 reductase gene of the tomato pathogenic fungusSeptoria lycopersici and its complementation of theerg-3 mutation ofNeurospora crassa

  • K. Aparna
  • Robert W. Sandrock
  • Durgadas P. Kasbekar


We have cloned theerg-3 gene, which encodes the ergosterol biosynthetic enzyme sterol C-14 reductase, from the tomato pathogenic fungusSeptoria lycopersici. Its nucleotide sequence, reported here, encodes a 512-amino-acid polypeptide with 54% sequence identity to sterol C-14 reductase ofNeurospora crassa. TheSeptoria gene complemented the pisatin-sensitive, tomatine-resistant and female-sterile phenotypes of aNeurospora erg-3 mutant.


Septoria lycopersici Neurospora crassa sterol C-14 reductase erg-3 pisatin tomatine 


  1. Arneson P. A. and Durbin R. D. 1967 Hydrolysis of tomatine bySeptoria lycopersici: A detoxification mechanism.Phytopathology 57, 1358–1360.Google Scholar
  2. Arneson P. A. and Durbin R. D. 1968a The sensitivity of fungi to α-tomatine.Phytopathology 58, 536–537.Google Scholar
  3. Arneson P. A. and Durbin R. D. 1968b Studies on the mode of action of tomatine as a fungitoxic agent.Plant Physiol. 43, 683–686.PubMedCrossRefGoogle Scholar
  4. Bruchez J. J. P., Eberle J. and Russo V. E. A. 1993a Regulatory sequences involved in the translation ofNeurospora crassa mRNA: Kozak sequences and stop codons.Fungal Genet. Newsl. 40, 85–88.Google Scholar
  5. Bruchez J. J. P., Eberle J. and Russo V. E. A. 1993b Regulatory sequences in the transcription ofNeurospora crassa genes: CAAT box, TATA box, introns, poly(A) tail formation sequences.Fungal Genet. Newsl. 40, 89–96.Google Scholar
  6. Cogoni C., Irelan J. T., Schumacher M., Schmidhauser T. J., Selker E. U. and Macino G. 1996 Transgene silencing of theal-1 gene in vegetative cells ofNeurospora is mediated by a cytoplasmic effector and does not depend on DNA: DNA interaction or DNA methylation.EMBO J. 15, 3153–3163.PubMedGoogle Scholar
  7. Davis R. H. and de Serres J. F. 1970 Genetic and microbiological research techniques forNeurospora crassa.Meth. Enzymol. A17, 79–143.CrossRefGoogle Scholar
  8. Ford J. E., McCance D. J. and Drysdale R. B. 1977 The detoxification of α-tomatine byFusarium oxysporum f. sp.lycopersici.Phytochemistry 16, 545–546.CrossRefGoogle Scholar
  9. Grotelueschen J. and Metzenberg R. L. 1995 Some property of the nucleus determines the competence ofNeurospora crassa for transformation.Genetics 139, 1545–1551.PubMedGoogle Scholar
  10. Lai M. H., Bard M., Pierson C. A., Alexander J. F., Goebl M., Carter G. T. and Kirsch D. R. 1994 The identification of a gene family in theSaccharomyces cerevisiae ergosterol biosynthesis pathway.Gene 140, 41–49.PubMedCrossRefGoogle Scholar
  11. Lorenz R. T. and Parks L. W. 1992 Cloning, sequencing and disruption of the gene encoding sterol C-14 reductase inSaccharomyces cerevisiae.DNA Cell Biol. 11, 685–692.PubMedGoogle Scholar
  12. Maniatis T., Fritsch E. F. and Sambrook J. 1982Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor.Google Scholar
  13. Marcireau C., Guyonnet D. and Karst F. 1992 Construction and growth properties of a yeast strain defective in sterol 14-reductase.Curr. Genet. 22, 267–272.PubMedCrossRefGoogle Scholar
  14. Miao V. P. W., Rountree M. R. and Selker E. U. 1995 Ectopic integration of transforming DNA is rare amongNeurospora transformants selected for gene replacement.Genetics 139, 1533–1544.PubMedGoogle Scholar
  15. Pandit N. N. and Russo V. E. A. 1992 Reversible inactivation of a foreign gene,hph, during the asexual cycle inNeurospora crassa transformants.Mol. Gen. Genet. 234, 412–422.PubMedCrossRefGoogle Scholar
  16. Papavinasasundaram K. G. and Kasbekar D. P. 1993 Pisatin resistance inDictyostelium discoideum andNeurospora crassa: comparison of mutant phenotypes.J. Gen. Microbiol. 139, 3035–3041.PubMedGoogle Scholar
  17. Papavinasasundaram K. G. and Kasbekar D. P. 1994 TheNeurospora crassa erg-3 gene encodes a protein with sequence homology to both yeast sterol C-14 reductase and chicken lamin B receptor.J. Genet. 73, 33–41.CrossRefGoogle Scholar
  18. Pegg G. F. and Woodward S. 1986 Synthesis and metabolism of α- tomatine in tomato isolines in relation to resistance toVerticillium albo-atrum.Physiol. Plant Pathol. 28, 187–201.Google Scholar
  19. Perkins D. D., Radford A., Newmeyer D. and Bjorkman M. 1982 Chromosomal loci ofNeurospora crassa.Microbiol. Rev. 46, 426–570.PubMedGoogle Scholar
  20. Sandrock R. W., DellaPenna D. and VanEtten H. D. 1995 Purification and characterization of β2-tomatinase, an enzyme involved in the degradation of α-tomatine, and isolation of gene encoding β2-tomatinase fromSeptoria lycopersici.Mol. Plant-Microb. Interact. 8, 960–970.Google Scholar
  21. Sanger F., Nicklen S. and Coulson A. R. 1977 DNA sequencing with chain-terminating inhibitors.Proc. Natl. Acad. Sci. USA 74, 5463–5467.PubMedCrossRefGoogle Scholar
  22. Schlosser E. 1975 Role of saponins in antifungal resistance IV. Tomatine-dependent development of species ofAlternaria on tomato fruits.Acta Phytopathol. 10, 77–87.Google Scholar
  23. Sengupta S., Bhavani Prasanna T. and Kasbekar D. P. 1995 Sterol 14,15 reductase (erg-3) mutants switch the phenotype ofNeurospora crassa from sensitivity to the tomato saponin α- tomatine to sensitivity to the pea phytoalexin pisatin.Fungal Genet. Newsl. 42, 71–72.Google Scholar
  24. Smith S. 1995 Cloning and sequence analysis of anERG24 homolog fromSchizosaccharomyces pombe.Gene 155, 139–140.PubMedCrossRefGoogle Scholar
  25. Sweigard J. and VanEtten H. D. 1987 Reduction in pisatin sensitivity ofAphanomyces euteiches by polar lipid extracts.Phytopathology 77, 771–775.CrossRefGoogle Scholar
  26. Verhoeff K. and Liem J. I. 1975 Toxicity of tomatine toBotrytis cinerea in relation to latency.Phytopathol. Z. 82, 333–338.CrossRefGoogle Scholar

Copyright information

© Indian Academy of Sciences 1998

Authors and Affiliations

  • K. Aparna
    • 1
  • Robert W. Sandrock
    • 2
  • Durgadas P. Kasbekar
    • 1
  1. 1.Centre for Cellular and Molecular BiologyHyderabadIndia
  2. 2.Department of Plant PathologyUniversity of ArizonaTucsonUSA

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