In Vitro Medium Composition and Culture Method Affecting Masspropagation of Osmunda japonica Thunb. Prothalli
2009
Shin, S.L., Chungbuk National University, Cheongju, Republic of Korea | Lee, C.H., Chungbuk National University, Cheongju, Republic of Korea
Present studies were carried out to investigate the effects of medium composition and cultural methods on the mass micropropagation of prothalli in Osmunda japonica Thumb. Prothallus proliferation was promoted on the media with low nutrient levels, such as Knop or one-eight strength MS medium. However, nutrient deficiency symptoms appeared during prothallus growth on Knop medium. To examine proper medium for good prothallus growth, various nitrogen sources and different concentration levels were employed using one-eight strength MS medium. The addition of 6 mM nitrogen resulted in best proliferation regardless of nitrogen sources. At the same level of nitrogen supplied, nitrate nitrogen was better than mixture of NH₄+:NO₃- at 1:2 ratio. Ammonia nitrogen retarded prothallus growth in this species. The multiplication of prothalli was compared between homogenized culture of chopped prothallus colony and culture of divided colony on modified one-eight strength MS medium (6 mM NO₃-, 3% sucrose, pH 5.8, 0.8% agar), and the former was better in proliferation through vigorous gemma formaton. So the homogenized prothalli were cultured on solid media containing agar concentation upto 1.2%, as well as on liquid media. Vigorous muliplication was observed on liquid culture with no glassy prothalli and no variation in form and frequency of sexual organ formation. Shaking liquid culture resulted in retarded growth of prothalli due to dying of prothalli tissue. In conclusion, recommended cultural method for mass micropropagation for O. japonica is stationary liquid culture using modified one-eight strength MS medium containing 6 mM of NO₃- as nitrogen source.
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