In vitro culture of Drynaria fortunei, a fern species source of Chinese medicine “Gu-Sui-Bu”

  • H.-C. Chang
  • D. C. Agrawal
  • C.-L. Kuo
  • J.-L. Wen
  • C.-C. Chen
  • H.-S. TsayEmail author
IAPB March/April 07


This study reports spore germination, early gametophyte development and change in the reproductive phase of Drynaria fortunei, a medicinal fern, in response to changes in pH and light spectra. Germination of D. fortunei spores occurred on a wide range of pH from 3.7 to 9.7. The highest germination (63.3%) occurred on ½ strength Murashige and Skoog basal medium supplemented with 2% sucrose at pH 7.7 under white light condition. Among the different light spectra tested, red, far-red, blue, and white light resulted in 71.3, 42.3, 52.7, and 71.0% spore germination, respectively. There were no morphological differences among gametophytes grown under white and blue light. Elongated or filamentous but multiseriate gametophytes developed under red light, whereas under far-red light gametophytes grew as uniseriate filaments consisting of mostly elongated cells. Different light spectra influenced development of antheridia and archegonia in the gametophytes. Gametophytes gave rise to new gametophytes and developed antheridia and archegonia after they were transferred to culture flasks. After these gametophytes were transferred to plastic tray cells with potting mix of tree fern trunk fiber mix (TFTF mix) and peatmoss the highest number of sporophytes was found. Sporophytes grown in pots developed rhizomes.


Gametophyte Medicinal fern Micropropagation Spore germination Sporophyte 



The authors gratefully acknowledged a research grant (NSC 95-2317-B-039-002) from the National Science Council, Taiwan.


  1. Anonymous. Flora of Taiwan, Vol.1, 2nd edn., Taipei: National Science Council of Taiwan; 1994:484–485.Google Scholar
  2. Anonymous. Pharmacopoeia Commission of People Republic of China (ChPC), Vol. 1. China: Chemical Industry Press; 2005:179–180.Google Scholar
  3. Camloh, M. Spore germination and early gametophyte development of Platycerium bifurcatum. Am. Fern J. 83:124–132; 1993.CrossRefGoogle Scholar
  4. Chang, E. J.; Lee, W. J.; Cho, S. H.; Choi, S. W. Proliferative effects of flavan-3 and propelargonidina from rhizomes of Drynaria fortunei on MCF-7 and osteblastic cells. Arch. Pharm. Res. 26:620–630; 2003.PubMedGoogle Scholar
  5. Conway, E. The autecology of bracken [Pteridium aquililinum (L.) Kuhn.]: The germination of spore, and the development of the prothallus and the young sporophyte. Proc. R. Soc. Edinb. 63B:625–643; 1949.Google Scholar
  6. Douglas, G. E. An investigation into the growth, development and ultra structure of fern gametophytes in existing and novel culture systems. PhD dissertation, University of Manchester, U.K; 1994.Google Scholar
  7. Edwards, M. E. Carbon dioxide and ethylene control of spore germination in Onoclea sensibilis L. Plant Physiol. 59:756–758; 1977.PubMedGoogle Scholar
  8. Edwards, M. E.; Miller, J. H. Growth regulation by ethylene in fern gametophytes. II. Inhibition of cell division. Am. J. Bot. 59:450–457; 1972a.CrossRefGoogle Scholar
  9. Edwards, M. E.; Miller, J. H. Growth regulation by ethylene in fern gametophytes. III. Inhibition of spore germination. Am. J. Bot. 59:458–465; 1972b.CrossRefGoogle Scholar
  10. Hevly, R. H. Adaptations of Cheilanthoid ferns to desert environments. J. Ariz. Acad. Sci. 2:164–175; 1963.Google Scholar
  11. Jeong, J. C.; Lee, J. W.; Yoon, C. H.; Lee, Y. C.; Chung, K. H.; Kim, M. G.; Kim, C. H. Stimulative effects of Drynariae rhizoma extracts on the proliferation and differentiation of osteoblastic MC3T3-E1 cells. J. Ethnopharmacol. 96:489–95; 2005.PubMedCrossRefGoogle Scholar
  12. Lin, C. Y.; Sun, J. S.; Sheu, S. Y.; Lin, F. H.; Wang, Y. J.; Chen, L. T. The effect of Chinese medicine on bone cell activities. Am. J. Chin. Med. 30:271–285; 2002.PubMedCrossRefGoogle Scholar
  13. Ma, K. C.; Zhu, T. Y.; Wang, F. X. Stimulatory effects of gu-sui-bu (Drynaria baronii) injection on chick embryo bone primordium calcification in vitro. Am. J. Chin. Med. 24: 77–82; 1996.PubMedCrossRefGoogle Scholar
  14. Mohr, H. Die Abhangigkeit des protonemawachstums und der protonemapolaritat bei farnen vom licht. Planta 47:127–158; 1956.CrossRefGoogle Scholar
  15. Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15:473–497; 1962.CrossRefGoogle Scholar
  16. Pajaron, S.; Pangua, E.; Garcia-Alvarez, L. Sexual expression and genetic diversity in populations of Cryptpgramma crispa (Pteridaceae). Am. J. Bot. 86:964–973; 1999.PubMedCrossRefGoogle Scholar
  17. Raghavan, V. Developmental biology of fern gametophytes. Cambridge: Cambridge University Press; 1989:7–151.Google Scholar
  18. Raghavan, V. Germination of fern spores. Am. Sci. 80:176–185; 1992.PubMedGoogle Scholar
  19. SAS Institute Inc. SAS/STAT User’s Guide. Version 8.2, vol 2. USA: SAS Institute; 2001:943.Google Scholar
  20. Sheffield, E.; Douglas, G. E.; Hearne, S. J.; Huxham, S.; Wynn, J. M. Enhancement of fern spore germination and gametophyte growth in artificial media. Am. Fern J. 91:179–186; 2001.CrossRefGoogle Scholar
  21. Sun, J. S.; Theriault, B. L.; Anderson, G. I. The effect of Gu-Sui-Bu (Drynaria fortunei) on bone cell activity. Am. J. Chin. Med. 32:737–53; 2004.PubMedCrossRefGoogle Scholar
  22. Swami, P.; Raghavan, V. Control of morphogenesis in the gametophyte of a fern by light and growth hormones. Can. J. Bot. 58:1464–1473; 1980.Google Scholar
  23. Wayne, R.; Hepler, P. K. The role of calcium ions in phytochrome-mediated germination of spores of Onoclea sensibilis L. Planta 160:12–20; 1984.CrossRefGoogle Scholar
  24. Wayne, R.; Hepler, P. K. Red light stimulates an increase in intracellular calcium in the spores of Onoclea sensibilis L. Plant Physiol. 77:8–11; 1985.PubMedCrossRefGoogle Scholar
  25. Whittier, D. P. Spore germination and early gametophyte development in Stromatopteris. Am. Fern J. 89:142–148; 1999.CrossRefGoogle Scholar
  26. Whittier, D. P. The gametophyte of Diphasiastrum sitchense. Am. Fern J. 93:20–24; 2003.CrossRefGoogle Scholar
  27. Wong, R. W.; Rabie, A. B. Systemic effect of crude extract from rhizome of Drynaria fortunei on bone formation in mice. Phytother. Res. 20:313–315; 2006.PubMedCrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2007

Authors and Affiliations

  • H.-C. Chang
    • 1
  • D. C. Agrawal
    • 2
  • C.-L. Kuo
    • 3
  • J.-L. Wen
    • 1
  • C.-C. Chen
    • 3
  • H.-S. Tsay
    • 2
    Email author
  1. 1.Institute of Chinese Pharmaceutical ScienceChina Medical UniversityTaichungTaiwan
  2. 2.Graduate Institute of BiotechnologyChaoyang University of TechnologyWufong, TaichungTaiwan
  3. 3.School of Chinese Medicine ResourcesChina Medical UniversityTaichungTaiwan

Personalised recommendations