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Isolation and Characterization of Double Deletion Mutants of nsdD and veA or nsdC, the Genes Positive Regulating Sexual Development of Aspergillus nidulans

Aspergillus nidulans에서 유성분화 능동조절 유전자 nsdD와 veA 또는 nsdC의 이중결실돌연변이의 분리 및 분석

  • Kim, Lee-Han (Division of Life Sciences, Wonkwang University) ;
  • Kim, Ha-Eun (Division of Life Sciences, Wonkwang University) ;
  • Han, Dong-Min (Division of Life Sciences, Wonkwang University)
  • Received : 2013.12.10
  • Accepted : 2013.12.21
  • Published : 2013.12.31

Abstract

Three genes, nsdD, nsdC, and veA are known to be necessary for positive regulating sexual development of Aspergillus nidulans. Since the mutants of those genes hardly form fruiting bodies in heterokaryons constructed by cross between two of them, it is difficult to isolate double mutants. In this work, double mutants of ${\Delta}nsdD$ ${\Delta}veA$ and ${\Delta}nsdD$ ${\Delta}nsdC$ were isolated using the characteristic of the nsdD deletion mutant that it could develop mature cleistothecia in hypoxic and low temperature culture condition. According to the phenotypes of double mutants, the nsdD gene controls the apical growth independently with veA or nsdC. Deletion of veA or nsdC was epistatic to nsdD deletion for pigment production. Conidia formation in submerged culture with lactose as sole carbon source was observed in ${\Delta}nsdD$ ${\Delta}nsdC$ double mutant implicating it to be unique phenotype of nsdC deletion.

Aspergillus nidulans에는 유성분화를 능동조절하는 nsdD, nsdC 그리고 veA의 3 유전자가 알려져 있다. 이들 유전자의 돌연변이들은 각각을 서로 교배한 이핵체에 cleistothecia를 형성하지 못하기 때문에 이중돌연변이의 분리가 어려웠다. 본 연구에서는 ${\Delta}nsdD$ 돌연변이가 저산소 및 저온조건에서 성숙한 cleistothecia를 형성하는 것을 이용하여 ${\Delta}nsdD$${\Delta}veA$${\Delta}nsdD$${\Delta}nsdC$의 이중돌연변이를 분리하였다. 이중돌연변이의 형질을 분석한 결과 nsdD 유전자는 veA 또는 nsdC와 독립적으로 정단성장에 관여하는 것으로 보였다. 색소생성형질은 veA나 nsdC가 nsdD에 대해 상위에 있는 것으로 나타났으며, lactose가 탄소원인 액체배지에서 무성포자를 생성하는 형질이 이중돌연변이에도 보임에 따라 이 형질은 nsdC의 고유한 특성으로 사료된다.

Keywords

References

  1. Bayram, O., Krappmann, S., Ni, M., Bok, J.W., Helmstaedt, K., Valerius, O., Braus-Stromeyer, S., Kwon, N.J., Keller, N.P., Yu, J.H., and et al. 2008. VelB/VeA/LaeA complex coordinates light signal with fungal development and secondary metabolism. Science 320, 1504-1506. https://doi.org/10.1126/science.1155888
  2. Calvo, A.M. 2008. The VeA regulatory system and its role in morphological and chemical development in fungi. Fungal Genet. Biol. 45, 1053-1061. https://doi.org/10.1016/j.fgb.2008.03.014
  3. Cary, J.W., Harris-Coward, P.Y., Ehrlich, K.C., Mack, B.M., Kale, S.P., Larey, C., and Calvo, A.M. 2012. NsdC and NsdD affect Aspergillus flavus morphogenesis and aflatoxin production. Eukaryot. Cell 11, 1104-1011. https://doi.org/10.1128/EC.00069-12
  4. Han, D.M., Han, Y.J., Lee, Y.H., Jahng, K.Y., Jahng, S.H., and Chae, K.S. 1990. Inhibitory conditions of asexual development and their application for the screening of mutants defective in sexual development. Kor. J. Mycol. 18, 225-232.
  5. Han, D.M., Han, Y.J., Kim, J.H., Jahng, K.Y., Chung, Y.S., Chung, J.H., and Chae, K.S. 1994. Isolation and characterization of NSD mutants in Aspergillus nidulans. Kor. J. Genet. 15, 1-7.
  6. Han, K.H., Han, K.Y., Yu, J.H., Chae, K.S., Jahng, K.Y., and Han, D.M. 2001. The nsdD gene encodes a putative GATA-type transcription factor necessary for sexual development of Aspergillus nidulans. Mol. Microbiol. 41, 299-309. https://doi.org/10.1046/j.1365-2958.2001.02472.x
  7. Han, K.H., Lee, D.B., Kim, J.H., Kim, M.S., Han, K.Y., Kim, W.S., Park, Y.S., Kim, H.B., and Han, D.M. 2003. Environmental factors affecting development of Aspergillus nidulans. J. Microbiol. 41, 34-40.
  8. Kim, H.S., Han, K.Y., Kim, K.J., Han, D.M., Jahng, K.Y., and Chae, K.S. 2002. The veA gene activates sexual development in Aspergillus nidulans. Fungal Genet. Biol. 37, 72-80. https://doi.org/10.1016/S1087-1845(02)00029-4
  9. Kim, H.R., Chae, K.S., Han, K.H., and Han, D.M. 2009. The nsdC gene encoding a putative $C_2H_2$-type transcription factor is a key activator of sexual development in Aspergillus nidulans. Genetics 182, 771-783. https://doi.org/10.1534/genetics.109.101667
  10. Mooney, J.L. and Yager, L.N. 1990. Light is required for conidiation in Aspergillus nidulans. Genes Dev. 4, 1473-1482. https://doi.org/10.1101/gad.4.9.1473
  11. Park, J.S. and Han, D.M. 2013. Isolation and characterization of the gtfA gene encoding GAL4-like transcription factor in Aspergillus nidulans. Kor. J. Microbiol. 49, 8-16. https://doi.org/10.7845/kjm.2013.020
  12. Yin, W. and Keller, N.P. 2011. Transcriptional regulatory elements in fungal secondary metabolites. J. Microbiol. 49, 329-339. https://doi.org/10.1007/s12275-011-1009-1
  13. Zonneveld, B.J.M. 1977. Biochemistry and ultrastructure of sexual development of Aspergillus, pp. 59-80. In Smith, J.E. and Pateman, J.A. (eds.), Genetics and Physiology of Aspergillus. Academic Press, London, UK.