DOI QR코드

DOI QR Code

Optimization of Protoplast Preparation and Regeneration of a Medicinal Fungus Antrodia cinnamomea

  • Wu, Jyun-De (Department of Life Science, National Dong Hwa University) ;
  • Chou, Jyh-Ching (Department of Natural Resources and Environmental Studies, National Dong Hwa University)
  • Received : 2019.08.19
  • Accepted : 2019.10.27
  • Published : 2019.12.01

Abstract

Antrodia cinnamomea is a unique medicinal fungus in Taiwan. It has been found rich in some pharmacologically active compounds for anti-cancer, hangover, and immune regulation etc. With the in-depth study of these components, it would be interesting and important to establish a molecular system for basic studies of A. cinnamomea. Thus, we would like to set up a foundation for this purpose by studying the A. cinnamomea protoplast preparation and regeneration. Firstly, we studied the optimization method of protoplast preparation of A. cinnamomea, and found various factors that may affect the yield during protoplast preparation, such as mycelial ages, pH values, and osmotic stabilizers. Secondly, in the regeneration of protoplasts, we explored the effects of various conditions on the regeneration of protoplasts, including different media and osmotic pressure. In addition, we found that citrate buffer with pH value around 3 dramatically increased the regeneration of protoplasts of A. cinnamomea, and provided a set of regeneration methodology for A. cinnamomea.

Keywords

References

  1. Hseu YC, Chang WC, Hseu YT, et al. Protection of oxidative damage by aqueous extract from Antrodia camphorata mycelia in normal human erythrocytes. Life Sci. 2002;71(4):469-482. https://doi.org/10.1016/S0024-3205(02)01686-7
  2. Song TY, Yen GC. Antioxidant properties of Antrodia camphorata in submerged culture. J Agric Food Chem. 2002;50(11):3322-3327. https://doi.org/10.1021/jf011671z
  3. Chen C-H, Yang S-W, Shen Y-C. New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. J Nat Prod. 1995;58(11):1655-1661. https://doi.org/10.1021/np50125a002
  4. Lee IH, Huang RL, Chen CT, et al. Antrodia camphorata polysaccharides exhibit anti-hepatitis B virus effects. FEMS Microbiol Lett. 2002;209(1):63-67. https://doi.org/10.1111/j.1574-6968.2002.tb11110.x
  5. Chang TT, Chou WN. Antrodia cinnamomum sp. nov. on Cinnamomum kanehirai in Taiwan. Mycol Res. 1995;99(6):756-758. https://doi.org/10.1016/S0953-7562(09)80541-8
  6. Geethangili M, Tzeng YM. Review of pharmacological effects of Antrodia camphorata and its bioactive compounds. Evidence-Based Compl Alt Med. 2011;2011:1. https://doi.org/10.1093/ecam/nep108
  7. Du YC, Wu TY, Chang FR, et al. Chemical profiling of the cytotoxic triterpenoid-concentrating fraction and characterization of ergostane stereoisomer ingredients from Antrodia camphorata. J Pharm Biomed Anal. 2012;58:182-192. https://doi.org/10.1016/j.jpba.2011.09.007
  8. Lin JY, Wu TZ, Chou JC. In vitro induction of fruiting body in Antrodia cinnamomea - A medicinally important fungus. Botan Stud. 2006;47:267-272.
  9. Chen ECF, Agrawal DC, Wang HY, et al. T-DNA activation-tagged mycelia producing fruiting bodies-specific triterpenoids in Antrodia cinnamomea - A medicinal fungus. Mycosphere. 2016;7(5):602-611. https://doi.org/10.5943/mycosphere/7/5/6
  10. Chu YC, Yang RM, Chang TT, et al. Fructification of Antrodia cinnamomea was strain dependent in malt extract media and involved specific gene expression. J Agric Food Chem. 2010;58(1):257-261. https://doi.org/10.1021/jf902793e
  11. Eeckhaut T, Lakshmanan PS, Deryckere D, et al. Progress in plant protoplast research. Planta. 2013;238(6):991-1003. https://doi.org/10.1007/s00425-013-1936-7
  12. Peberdy JF. Fungal protoplasts: isolation, reversion, and fusion. Annu Rev Microbiol. 1979;33(1):21-39. https://doi.org/10.1146/annurev.mi.33.100179.000321
  13. Gow NAR, Latge JP, Munro CA. The fungal cell wall: structure, biosynthesis, and function. Microbiol Spectr. 2017;5(3). doi:10.1128/microbiolspec.FUNK-0035-2016
  14. Gallmetzer M, Burgstaller W, Schinner F. An optimized method for the isolation of protoplasts from Penicillium simplicissimum to produce sealed plasma membrane vesicles. Mycologia. 1999;91(1):206-212. https://doi.org/10.2307/3761209
  15. Biot-Pelletier D, Martin VJJ. Evolutionary engineering by genome shuffling. Appl Microbiol Biotechnol. 2014;98(9):3877-3887. https://doi.org/10.1007/s00253-014-5616-8
  16. Lu MC, El-Shazly M, Wu TY, et al. Recent research and development of Antrodia cinnamomea. Pharmacol Ther. 2013;139(2):124-156. https://doi.org/10.1016/j.pharmthera.2013.04.001
  17. Feng H, Sun Z, Li H, et al. Preparation, purification and regeneration optimizing research of protoplasts from Rhizoctonia solani. Afr J Microbiol Res. 2012;6:3222-3230.
  18. Yu PW, Chang YC, Liou RF, et al. pks63787, a polyketide synthase gene responsible for the biosynthesis of benzenoids in the medicinal mushroom Antrodia cinnamomea. J Nat Prod. 2016;79(6):1485-1491. https://doi.org/10.1021/acs.jnatprod.5b00798
  19. Wubie A, Hu Y, Li W, et al. Factors analysis in protoplast isolation and regeneration from a Chalkbrood fungus, Ascosphaera apis. Int J Agric Biol. 2014;16:89-96.
  20. Komoda K, Naito S, Ishikawa M. Replication of plant RNA virus genomes in a cell-free extract of evacuolated plant protoplasts. Proc Natl Acad Sci USA. 2004;101(7):1863-1867. https://doi.org/10.1073/pnas.0307131101
  21. Fuchino K, Flardh K, Dyson P, et al. Cell-biological studies of osmotic shock response in Streptomyces spp. J Bacteriol. 2017;199:e00465-16.
  22. Rehman L, Su X, Guo H, et al. Protoplast transformation as a potential platform for exploring gene function in Verticillium dahliae. BMC Biotechnol. 2016;16(1):57. https://doi.org/10.1186/s12896-016-0287-4
  23. Geissenhoner A, Sievers N, Brock M, et al. Aspergillus nidulans DigA, a potential homolog of Saccharomyces cerevisiae Pep3 (Vps18), is required for nuclear migration, mitochondrial morphology and polarized growth. Mol Genet Genom. 2001;266(4):672-685. https://doi.org/10.1007/s00438-001-0589-6
  24. Veses V, Richards A, Gow NAR. Vacuoles and fungal biology. Curr Opin Microbiol. 2008;11(6):503-510. https://doi.org/10.1016/j.mib.2008.09.017
  25. Lu ZM, He Z, Li HX, et al. Modified arthroconidial inoculation method for the efficient fermentation of Antrodia camphorata ATCC 200183. Biochem Eng J. 2014;87:41-49. https://doi.org/10.1016/j.bej.2014.03.020
  26. Takeshita N. Coordinated process of polarized growth in filamentous fungi. Biosci Biotechnol Biochem. 2016;80(9):1693. https://doi.org/10.1080/09168451.2016.1179092

Cited by

  1. Establishment of a Genetic Transformation System in Guanophilic Fungus Amphichorda guana vol.7, pp.2, 2019, https://doi.org/10.3390/jof7020138
  2. Enhancement of antroquinonol production via the overexpression of 4-hydroxybenzoate polyprenyltransferase biosynthesis-related genes in Antrodia cinnamomea vol.184, 2019, https://doi.org/10.1016/j.phytochem.2021.112677