• Korean Journal of Plant Resources
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• v.32 no.6
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• pp.735-742
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• 2019

This study investigated a suitable method that could be applied for Asian chain fern [Woodwardia japonica (L. f.) Sm.] to propagate gametophytes and promote sporophyte formation. The gametophytes used in all experiments were obtained from germinated spores in vitro and were subcultured at 8-week intervals. The most appropriate media for gametophyte propagation was identified by culturing 300 mg of gametophyte in Murashige and Skoog (MS) basal medium (1/8, 1/4, 1/2, 1, 2), and Knop medium for 8 weeks. As a result, fresh weight of the gametophyte was increased by 56.7-fold on MS medium. Moreover, antheridium formation as well as gametophyte growth was improved on MS medium, especially. To improve the sporophyte formation ex vitro, 1.0 g of gametophyte was ground with distilled water and spread on eight combinations onto four different culture mediums, such as bed soil, peat moss, perlite and decomposed granite. Then generation and growth of sporophytes were investigated after cultivation for 10 weeks. As a result of this experiment, peat moss had a promotive effect of sporophyte formation at single-use and mixed culture soils. In particular, a mixture of bed soil, peat moss and perlite in a 1:1:1 ratio (v/v/v) led to the accelerated formation (782.5 ea/pot) and the frond growth of sporophytes. This included increases in length and width of fronds. However, promotive effect of gametophyte growth and sporophyte formation was not found at single-use and treatment with high ratio of bed soil.

• ALGAE
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• v.34 no.2
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• pp.153-162
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• 2019

Anthropogenic disturbances, including coastal habitat modification and climate change are threatening the stability of kelp beds, one of the most diverse and productive marine ecosystems. To test the effect of temperature and irradiance on the microscopic gametophyte and juvenile sporophyte stages of the rare kelp, Saccharina angustissima, from Casco Bay, Maine, USA, we carried out two sets of experiments using a temperature gradient table. The first set of experiments combined temperatures between $7-18^{\circ}C$ with irradiance at 20, 40, and $80{\mu}mol\;photons\;m^{-2}\;s^{-1}$. The second set combined temperatures of $3-13^{\circ}C$ with irradiance of 10, 100, and $200{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Over two separate 4-week trials, in 2014 and again in 2015, we monitored gametogenesis, the early growth stages of the gametophytes, and early sporophyte development of this kelp. Gametophytes grew best at temperatures of $8-13^{\circ}C$ at the lowest irradiance of $10-{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Light had a significant effect on both male and female gametophyte growth only at the higher temperatures. Temperatures of $8-15^{\circ}C$ and irradiance levels of $10-100{\mu}mol\;photons\;m^{-2}\;s^{-1}$ were conditions for the highest sporophyte growth. Sporophyte and male gametophyte growth was reduced at both temperature extremes-the hottest and coldest temperatures tested. S. angustissima is a unique kelp species known only from a very narrow geographic region along the coast of Maine, USA. The coupling of global warming with high light intensity effects might pose stress on the early life-history stages of this kelp, although, as an intertidal species, it could also be better adapted to temperature and light extremes than its subtidal counterpart, Saccharina latissima.

• 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.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.

• 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.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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• 2001.04a
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• pp.38-40
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• 2001

• Proceedings of the Plant Resources Society of Korea Conference
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• 2001.04a
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• pp.41-42
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• 2001

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.454-461
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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 Polystichum braunii (Spenn.) $F{\acute{e}}e$. The rate of spore germination and early prothalium development was high in Knop (41.2%), which had low mineral content. The optimal medium for prothallus propagation and sexual organ formation was 2MS medium (2% sucrose). Among the various mixtures of cultivation soil (bedding soil, peat moss, perlite, and decomposed granite), a mixture of bedding soil and decomposed granite at a ratio of 2:1 (v:v) had a positive effect on sporophyte formation (276.3 ea/$7.5m^2$). The most efficient conditions for promoting the growth of sporophytes were pots filled with only bedding soil.

• 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.