• 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.

• Korean Journal of Plant Resources
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• v.22 no.4
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• pp.337-342
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• 2009

Present studies were conducted to evaluate the effects of medium strength(MS and Hyponex), carbon sources and their concentrations, agar concentrations, and inoculation amounts on prothallus propagation of Pterdium aquilinum var. latiusculum(Desv.) Underw. ex Hell in vitro. The optimum MS medium strength for prothallus propagation was 2MS concentration. Phosphate source was most effective for prothallus growth of P. aquilinum var. latisculum. The addition of 1% sucrose or glucose to MS medium promoted prothallus multiplication. Growth of prothallus was not affected by agar concentration. Propagation of homogenized prothallus was vigorous even in liquid medium. Chopped gametophytes(100 and 200 mg) were inoculated on 250 ml ${\Delta}$flask with 100 mL of 2MS concentration medium and suspension culture was done at 100 rpm for 22 days. After 20 days, prothallus multiplication slowed down, so 100 mg of chopped prothalli is recommended for initial inoculation, since initial amount of inoculum did not affect subsequent prothallus multiplication. Consequently after 20 days of suspension culture, prothallus should be subcultured or transplanted outside of growing vessels.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.04a
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• pp.43-44
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• 2002

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.259-264
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• 2006

This experiment was conducted to investigate the effects of medium composition, physical condition and growth regulators on prothallus proliferation in 3 fern species belonging to Aspleniaceae (Asplenium incisum, Camptosorus sibiricus and Phyllitis scolopendrium). In all of the 3 species, the effective medium for prothallus proliferation was Murashige and Skoog's basal medium with 1% sucrose and pH 5.8. The optimum $NH_4^+:NO_3^-$ concentration ratio of protahllus proliferation was 20:40mM (1:2 ratio) in A. incisum and P. scolopendrium and 30:30mM (1:1 ratio) in C. sibiricus. However, agar content of medium did not affect the rate of prothallus proliferation. Among the selected 6 kinds of growth regulators (NAA, IAA, 2,4-D, BAP, kinetin and lip), cytokinins generally has a promotive effect, but auxins showed an inhibitory effect on prothallus growth.

• Horticultural Science & Technology
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• v.35 no.1
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• pp.46-58
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• 2017

This study was conducted to investigate the optimal conditions for spore germination, prothallus propagation, sporophyte formation and seedling growth in Microlepia strigosa (Thunb.) C. Presl. Spore germination and prothallus development were promoted by low concentrations of Knop medium nutrient solution. The optimal medium for prothallus propagation and antheridium formation was 2X MS medium with 3% sucrose. The activated charcoal content of the medium did not affect prothallus proliferation. Among the various combinations of culture soil (bedding soil, peat moss, perlite and decomposed granite), a mixture of bedding soil, peat moss and decomposed granite at a ratio of 1 : 1 : 1 (v : v : v) had a positive effect on sporophyte formation. The most efficient conditions for promoting the growth of whole plants (sporophyte seedlings) were 50 - cell plug trays filled with a mixture of bedding soil and decomposed granite at a 2 : 1 (v : v) ratio.

• 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.32 no.4
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• pp.290-295
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• 2019

This study was carried out to select the optimal condition for prothallus propagation and sporophyte formation of Leptogramma pozoi (Lag.) Ching subsp. mollissima (Fisch. ex Kunze) Nakaike and to provide basic data for mass production system. In the propagation, 300 mg of prothallus were inoculated in different kinds of medium [1/4, 1/2, Murashige and Skoog (MS), Knop], sucrose (0, 1, 2, 3, 4%), and cultured for 8 weeks. Sporophyte formation studies were carried out by inoculating blended prothallus into artificial soils. The soils were mixed with horticultural substrate, peatmoss, perlite, and decomposed granite at different ratios or only with the horticultural substrate. Then the prothallus was cultivated for 12 weeks to figure out the formation and growth of the sporophytes. The growth and development of prothalli were excellent on MS medium. According to medium components, prothallus growth was favorable in all treatments except 0% sucrose treatment and the highest in 2% sucrose. In the experiment of soil mixtures, sporophyte formation was the highest in the horticultural substrate:perlite 2:1 (v:v) and horticultural substrate:decomposed granite 2:1 (v:v) treatment, and the overall growth was good in the horticultural substrate:decomposed granite 2:1 (v:v) mixed soil.

• 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.

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.348-354
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• 2006

Homogenized prothallus of 6 species (Cyrtomium falcatum, Cyrtomium caryoptideum var. coreanum, Dryopteris varia, Asplenium incisum, Camptosous sibiricus and Phyllitis scolopendrium) were treated with gamma radiation or by chemical mutagenesis with EMS, NMU, $NaN_3$ and colchicine to assess their sensitivities for each treatments and also with the aim of inducing mutations. Generally, decrease of proliferation ratio was dose-dependent and time-dependent. Based on proliferation ratio, optimum dose of gamma irradiation was $5{\sim}10krad$ except in D. varia with 20krad. Optimum condition of EMS treatment was considered as 50mM for 3h and for NMU as $5{\sim}10mM\;for\;1{\sim}3h$. Optimum condition of $NaN_3$ treatment was considered as $0.5{\sim}1mM\;for\;1{\sim}3h$. For colchicine, there were significant differences between species as to the proliferation ratios of prothallus for each treatment.

• FLOWER RESEARCH JOURNAL
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• v.18 no.3
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• pp.179-185
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• 2010

This study was carried out to investigate the optimal culture conditions for gametophytes growth and sporophytes regeneration of Pyrrosia linearifolia in order to provide for the masspropagation system foundation of Pyrrosia linearifolia using their life cycle. Among many different media, 2MS medium was most effective in prothallus proliferation. Prothallus growth was promoted as the total concentration of nitrogen sources increased, and the best result was observed on 120 mM nitrogen. The best concentration of sucrose was 3%. The addition of 5~20 mM IAA, NAA, BA and kinetin promoted the propagation of prothallus. But 2iP demonstrated the most inhibitory effect on prothallus proliferation. Gametophytes shaking-cultured with liquid medium showed similar growth with solid medium and normal formation of reproductive organs. Shoot regeneration was most effective on 1/8MS medium, but growth was promoted on 1/2MS medium. For promotion of shoot regeneration and growth, the suitable concentrations of sucrose and $NaH_2PO_4$ were 1% and $50{\sim}100mg{\cdot}mL^{-1}$ in 1/8MS medium, respectively.