A methodology for large-scale Athyrium sheareri gametophyte proliferation and sporophyte production using tissue culture

  • Bo Kook Jang
  • Ju Sung Cho
  • Kyungtae Park
  • Cheol Hee LeeEmail author


Tissue culture methods using gametophytes are considered the easiest ways to mass-produce fern sporophytes. The aim of this study was to develop a practical propagation method for the ornamental fern, Athyrium sheareri. The gametophytes obtained from in vitro spore germination were used as experimental materials. We used the chopping method to investigate the culturing conditions for proliferating gametophytes and the blending method for evaluating the mass production of sporophytes in mixed soil. Gametophyte proliferation was determined via Knop medium, various concentrations of Murashige and Skoog (MS) basal medium (1, 1/2, 1/4), and media components (sucrose, nitrogen source, and activated charcoal). The fresh weight of the gametophytes increased by more than 24-fold in 1/2 MS medium. In addition, 1 g of gametophyte could produce a maximum of 255.3 sporophytes in a mixed soil of 7.5 cm2 area. Treating gametophytes with exogenous plant growth regulators promoted the formation and growth of sporophytes. The cultivated young sporophytes were acclimated and successfully grown in greenhouses. We developed a mass production protocol for A. sheareri sporophytes suitable for field application, which is expected to have commercial value.


Blending method Chopping method Fern propagation Gametophyte Gibberellic acid 



This study was financially supported by the project ‘Development of Mass Propagation Protocol for the Establishment of Application Base Using Forest Plants’ of Korea National Arboretum, Project No. KNA1-2-25, 16-3.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Division of Animal, Horticultural and Food SciencesChungbuk National UniversityCheongjuRepublic of Korea
  2. 2.Brain Korea 21 Center for Bio-Resource DevelopmentChungbuk National UniversityCheongjuRepublic of Korea

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