DOI QR코드

DOI QR Code

Nucleus-Selective Expression of Laccase Genes in the Dikaryotic Strain of Lentinula edodes

  • Ha, Byeongsuk (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Lee, Sieun (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Kim, Sinil (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Kim, Minseek (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Moon, Yoon Jung (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Song, Yelin (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University) ;
  • Ro, Hyeon-Su (Division of Applied Life Science and Research Institute of Life Sciences, Gyeongsang National University)
  • 투고 : 2017.09.18
  • 심사 : 2017.10.22
  • 발행 : 2017.12.01

초록

In mating of Lentinula edodes, dikaryotic strains generated from certain monokaryotic strains such as the B2 used in this study tend to show better quality of fruiting bodies regardless of the mated monokaryotic strains. Unlike B2, dikaryotic strains generated from B16 generally show low yields, with deformed or underdeveloped fruiting bodies. This indicates that the two nuclei in the cytoplasm do not contribute equally to the physiology of dikaryotic L. edodes, suggesting an expression bias in the allelic genes of the two nuclei. To understand the role of each nucleus in dikaryotic strains, we investigated single nucleotide polymorphisms (SNPs) in laccase genes of monokaryotic strains to reveal nuclear origin of the expressed mRNAs in dikaryotic strain. We performed reverse transcription PCR (RT-PCR) analysis using total RNAs extracted from dikaryotic strains (A5B2, A18B2, and A2B16) as well as from compatible monokaryotic strains (A5, A18, and B2 for A5B2 and A18B2; A2 and B16 for A2B16). RT-PCR results revealed that Lcc1, Lcc2, Lcc4, Lcc7, and Lcc10 were the mainly expressed laccase genes in the L. edodes genome. To determine the nuclear origin of these laccase genes, the genomic DNA sequences in monokaryotic strains were analyzed, thereby revealing five SNPs in Lcc4 and two in Lcc7. Subsequent sequence analysis of laccase mRNAs expressed in dikaryotic strains revealed that these were almost exclusively expressed from B2-originated nuclei in A5B2 and A18B2 whereas B16 nucleus did not contribute to laccase expression in A2B16 strain. This suggests that B2 nucleus dominates the expression of allelic genes, thereby governing the physiology of dikaryons.

키워드

참고문헌

  1. Casselton LA, Olesnicky NS. Molecular genetics of mating recognition in basidiomycete fungi. Microbiol Mol Biol Rev 1998;62:55-70.
  2. Brown AJ, Casselton LA. Mating in mushrooms: increasing the chances but prolonging the affair. Trends Genetic 2001; 17:393-400. https://doi.org/10.1016/S0168-9525(01)02343-5
  3. Kües U. Life history and developmental processes in the basidiomycete Coprinus cinereus. Microbiol Mol Biol Rev 2000;64:316-53. https://doi.org/10.1128/MMBR.64.2.316-353.2000
  4. Leatham GF. Extracellular enzymes produced by the cultivated mushroom Lentinus edodes during degradation of a lignocellulosic medium. Appl Environ Microbiol 1985;50:859-67.
  5. Makkar RS, Tsuneda A, Tokuyasu K, Mori Y. Lentinula edodes produces a multicomponent protein complex containing manganese (II)-dependent peroxidase, laccase and $\beta$-glucosidase. FEMS Microbiol Lett 2001;200:175-9.
  6. Silva EM, Martins SF, Milagres AM. Extraction of manganese peroxidase produced by Lentinula edodes. Bioresour Technol 2008;99:2471-5. https://doi.org/10.1016/j.biortech.2007.04.064
  7. Samils N, Oliva J, Johannesson H. Nuclear interactions in a heterokaryon: insight from the model Neurospora tetrasperma. Proc Biol Sci 2014;281:20140084. https://doi.org/10.1098/rspb.2014.0084
  8. Ha BS, Kim S, Ro HS. Isolation and characterization of monokaryotic strains of Lentinula edodes showing higher fruiting rate and better fruiting body production. Mycobiology 2015;43:24-30. https://doi.org/10.5941/MYCO.2015.43.1.24
  9. Matsumoto T, Kitamoto Y. Induction of fruit-body formation by water-flooding treatment in sawdust cultures of Lentinus edodes. Trans Mycol Soc Jpn 1987;28:437-43.
  10. Floudas D, Binder M, Riley R, Barry K, Blanchette RA, Henrissat B, Martinez AT, Otillar R, Spatafora JW, Yadav JS, et al. The Paleozoic origin of enzymatic lignin decomposition reconstructed from 31 fungal genomes. Science 2012;336: 1715-9. https://doi.org/10.1126/science.1221748
  11. Shim D, Park SG, Kim K, Bae W, Lee GW, Ha BS, Ro HS, Kim M, Ryoo R, Rhee SK, et al. Whole genome de novo sequencing and genome annotation of the world popular cultivated edible mushroom, Lentinula edodes. J Biotechnol 2016;223:24-5. https://doi.org/10.1016/j.jbiotec.2016.02.032
  12. Park M, Kim M, Kim S, Ha B, Ro HS. Differential expression of laccase genes in Pleurotus ostreatus and biochemical characterization of laccase isozymes produced in Pichia pastoris. Mycobiology 2015;43:280-7. https://doi.org/10.5941/MYCO.2015.43.3.280
  13. Park JW, Kang HW, Ha BS, Kim SI, Kim S, Ro HS. Straindependent response to $Cu^{2+}$ in the expression of laccase in Pycnoporus coccineus. Arch Microbiol 2015;197:589-96. https://doi.org/10.1007/s00203-015-1090-7
  14. Murata H, Ohta A, Yamada A, Horimai Y, Katahata S, Yamaguchi M, Neda H. Monokaryotic hyphae germinated from a single spore of the ectomycorrhizal basidiomycete Tricholoma matsutake. Mycoscience 2015;56:287-92. https://doi.org/10.1016/j.myc.2014.08.004