• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.44-44
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• 2019

This study aimed to develop a suitable method for inducing the proliferation of prothallus and producing sporophytes of rock polypody (Polypodium vulgare L.). The prothalli used in all experiments were obtained from spore germination and sub-cultured for 8-week intervals. The most appropriate media for prothallus propagation were investigated by culturing 300 mg of prothallus in MS ($1/4{\times}$, $1/2{\times}$, $1{\times}$, and $2{\times}$ strength) medium and in Knop medium for 8 weeks. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\pm}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark). Fresh weight of prothalli was 4.8 g on $1{\times}$ MS, 4.5 g on $1/2{\times}$ MS and 4.3 g on 1/4 MS medium. To select a suitable soil combination for sporophyte formation, 1.0 g of prothallus was ground with distilled water, spread in five combinations onto different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and then cultivated for 13 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). The results showed that a mixture containing a 2:1 (v:v) ratio of horticultural substrate and perlite, increased sporophyte formation to 462.5 sporophytes per pot (7.5 cm2). The other soil substrates produced from 314.5 to 405.3 sporophytes per pot. Therefore, our results will provide conditions suitable for mass production of Polypodium vulgare L.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.43-43
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• 2019

Deparia pycnosora (Christ) M. Kato is a fern used as ornamental plant. In addition, it is called "Teol-go-sa-ri" in Korean name. The aim of this study was to develop a practical propagation method of D. pycnosora using tissue culture technique. Prothallus obtained from spore germination was the used as experiment materials. The prothalli (300 mg) used in all experiments were sub-cultured for 8-week intervals. The most suitable media for prothallus propagation were identified by culturing 300 mg of prothalli in $1/4{\times}$, $1/2{\times}$, $1{\times}$, $2{\times}$ MS medium and in Knop medium for 8 weeks. Also, the prothalli were cultured by chopping with a scalpel. In addition, sucrose, activated charcoal, and total nitrogen source were added in different concentrations based on the culture medium selected. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\times}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark) in in vitro. The results showed that optimum was achieved prothallus fresh weight and development in $1{\times}$ MS medium. When other components were added to the basic $1{\times}$ MS medium, prothallus propagation was maximized in $1{\times}$ MS medium supplemented with 2% sucrose, 0.2% activated charcoal, and 60 mM total nitrogen. To select a suitable soil mixture for sporophyte formation, 1.0 g of prothallus was blended with distilled water, spread on five combinations of different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and cultivated for 12 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). As a results, horticultural substrate alone, 2:1 (v:v) mixtures of horticultural substrate and perlite, and 2:1 mixtures of horticultural substrate and decomposed granite induced 208.0, 201.3 and 248.8 sporophytes per pot, respectively. Therefore, this result could provide a practical mass propagation method of D. pycnosora

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.252-258
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• 2006

The most effective conditions of In vitro culture and ex vitro sporophyte formation from prothallus were studied for mass propagation of Dyropteris varia. The most effective medium of prothallus proliferation was Murashige and Skoog's basal medium supplemented with 10:50mM of $NH_4^+:NO_3^-$ and 2% sucrose. The optimum pH level was 5.8 and prothallus growth was promoted on medium containing $0.6{\sim}0.8%$ agar. Almost of the tested growth regulators (NAA, IAA, 2,4-D, BAP, kinetin and 2ip) were inhibitory in prothallus proliferation as the concentration of growth regulators became higher. The highest number of sporophytes was obtained by transplanting prothallus on compost only than on any other soil compositions. Sporophyte formation was promoted remarkably by soaking prothallus with $100{\mu}M\;GA_3$ for 3 hours.

• FLOWER RESEARCH JOURNAL
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• v.17 no.2
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• pp.114-120
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• 2009

The most effective conditions of in vitro culture were studied for mass propagation of Pteris cretica 'Wilsonii'. Spores of the species germinated within 7 weeks. The greatest proliferation was obtained with Knop and Hyponex media, but growth was more effective in Hyponex medium. MS medium induced necrosis of prothalli in all strength of nitrogen and sucrose except in case of 0% sucrose. Hyponex medium supplemented with 1% sucrose and 0.6% agar promoted propagation and growth of prothalli. In Hyponex medium, optimal inoculation method was homogenization, but in MS medium dividing colonies of prothalli was more effective. Culturing on solid medium was more effective than liquid culture method. Liquid culture induced necrosis of prothalli. Shaking cultured prothalli showed good growth, but propagation was inhibited compared to those cultured on solid medium.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.131-141
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• 2017

This study was conducted to investigate in vitro mass propagation methods suitable for each growth stage of A. aristata (G. Forst.) Tindale, from spore germination to sporophyte formation. Among spores germinated in $1/8-1{\times}MS$ medium and Knop medium, Knop medium yielded the highest germination percentage (87.1%). We cultured prothalli obtained from germinating spores for 8 weeks on media with different concentrations of sucrose and active carbon, as well as different concentrations and ratios of nitrogen, to select a suitable growth medium. A. aristata (G. Forst.) Tindale prothalli grew most actively in MS medium with 3% sucrose and 20 : 40 mM of $NH_4Cl$ and $KNO_3$ (total concentration of 60 mM). We investigated sporophyte formation according to soil type, finding that bedding soil mixed with perlite at a 2 : 1(v / v) ratio yielded the highest number of sporophytes per pot ($73.8/7.5{\times}7.5cm\;pot$). By contrast, when peat moss was used alone or mixed with other substrates, prothallus development and sporophyte formation were suppressed. Therefore, the most effective propagation method for A. aristata (G. Forst.) Tindale is to grow prothalli in MS medium and to induce sporophyte formation in a mixture of bedding soil and perlite (v / v = 2 : 1).

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.394-402
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• 2009

Adventitious shoots were induced from pinnae, petiole and rhizome in Pteris cretica 'Wilsonii' in order to develop the efficient mass propagation method, using in vitro culture. Only homogenized rhizome segments could regenerate young sporophytes. Efficient regeneration of multiple shoots was obtained on the one-eighth strength MS medium containing 1% sucrose, and $50mg{\cdot}L^{-1}$ $NaH_2PO_4$. To achieve higher rate of regeneration from rhizome segments, rhizome segments were exposed to growth regulators for 2 months and then subcultured on hormone-free medium. The greatest shoot regeneration was obtained by $1{\mu}M$ kinetin with $5{\mu}M$ NAA. BA was effective in formation of GGB (kind of meristems), but they showed low shoot regeneration rate. Plants obtained from present experiments were transplanted to examine good environmental conditions for acclimation. Juvenile plants obtained by the one-eighth strength MS medium showed highest survival rate and vigorous growth at the seedling stage.

• FLOWER RESEARCH JOURNAL
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• v.17 no.2
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• pp.93-100
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• 2009

This study was carried out to investigate the efficient in vitro mass propagation methods for juvenile sporophytes of Matteuccia struthiopteris. Chopped segments of pinnae, petiole and rhizome were cultured on 1/2MS with 0.1% activated charcoal. Among these explant sources only rhizome segments produced young sporophytes, regenerating vigorously on 1/2 MS medium. Adjusting sucrose concentration to 2% and supplement to $50mgL^{-1}$ $NaH_2PO_4$ in 1/2MS medium proved to be more efficient for plant regeneration. Various combinations of growth regulators such as kinetin, BA, NAA, and IBA were added to the growing media, and the best sporophyte regeneration was obtained by $1{\mu}M$ kinetin. The BA addition resulted in vigorous proliferation of meristematic tissues, but without differentiation to sporophytes. Three types of culture methods, solid using agar, liquid stationary, and liquid shaking culture, were employed with or without activated charcoal. The addition of 0.1% activated charcoal to modified 1/2MS media (2% sucrose, $50mgL^{-1}$ $NaH_2PO_4$, $1{\mu}M$ kinetin, pH 5.8 and 0.8% agar) yielded highest sporophyte regeneration in liquid shaking culture.