• Proceedings of the Plant Resources Society of Korea Conference
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• 2019.04a
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• pp.53-53
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• 2019

Cypripediums, popularly called lady's slippers or moccasin flowers, are the showiest and most sought after hardy terrestrial orchids, collected and grown by orchid and alpine plant enthusiasts alike. In Korea, 4 species of cypripedium are reported as Cypripedium japonicum, C. macranthos, C. guttatum, and C. calceolus. We had already reported the feasibilities of C. macranthos and C. guttatum with in vitro germination methods from immature seeds. The seeds of white lady's slipper orchid (Cypripedium macranthos Sw. alba) were collected 65 days after pollination in 2018. The green pods were sterilized with flame and sowed immediately on the POM(Phytomax orchid maintenance media(R), Sigma) supplemented with BAP 0, 0.5, 1.0 mg/L and NAA 0, 1, 2mg/L. The germination of seed was observed 90 days after sowing, and the plantlets were subcultured to the same media according to the size of the protocorm with 1~2, 2~3, 3~4, 5~6, 7~8mm. The time of the subculture to the new media seems to be critical factors of forming rhizoids which is the hairy root of the cypripediums. As a results, the protorms of the white lady's slipper orchid was successfully germinated in the POM media supplemented BAP 0.5 and NAA 1.0 mg/L. The roots and rhizoids were formed in 5~6mm protocorms subculture over 95% survival ratio. We also tried to subculture to liquid medium without activated charcoal, however the browning or malformation of the roots was observed in the root. The formation of shoots from the protocorm was effectively enhanced in the POM media with non-additives of plant growth regulators. These results indicate the possibility of high and stable production and practical industrialization of endangered white lady's slipper orchids.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.89-94
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• 2000

In order to induce the mutants of Gentiana axillariflora Leveille, nodes were cultured on Schenk and Hilderbrandt (SH) medium containing TDZ 2 mg/L, BAP 2mg/L, GA3 0.5 mg/L and 0.1 mg/L NAA and each mutagen of ethylmethanesulfate (EMS), colchicine, N-methyl-N-nitrosourea (MNU), and sodium azide (NaN$_3$) through filtration. Comparision of morphological characteristics and survival rate in each mutant plants differed depending on mutagen sources and their concentrations. When EMS were treated on nodes, the regenerated plants was thin and albino, regenerated shoots appeared 'erectoides type' and get twisted. The case of colchicine were treated on nodes, the survival rate was from 84% to 97% at ail concentration after 30days but the rate of survival was decreased about 50% at 200 $\mu$M after 60days. The treatment of NaN$_3$200 $\mu$M was not survived. The survival rate was extremely decreased in MNU treatment at 500$\mu$M, according to concentrations two types of leaf characteristic were obtained. Type I of leaf characteristic was modified from oblanceolate to oboid at leaf shape and type II of leaf characteristic was modified from light green to dark violet at leaf color. RAPD analysis was carried out to check the genetic modification of regenerated plants by mutagen treatments. Three polymorphic DNA fragments out of thirty-seven obtained by RAPDs were observed in regenerated plants using 5 decamer primers.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.1-6
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• 2009

A study was conducted to investigate in vitro micro propagation of four potato cultivars of Daese, Jasim, Chubaek and Haryeng in MS medium and PM medium (a medium containing half concentration of $NH_4NO_3$ and $KNO_3$, two fold concentration of $CaCl_2$ as compared to MS medium). During $1{\sim}2$ weeks of culture, Daese and Jasim showed better shoot elongation on the solid MS medium than Chubaek and Haryeng whereas Chubaek and Haryeng did better shoot elongation on the solid PM medium. But no difference was observed after 4 weeks of culture. As compared to shoot elongation on the solid medium, it was delayed at early stage of culture in the liquid medium without shaking. Shoot formation ratio of potato (above 4 cm of shoot length) began to increase significantly after 1 week of culture and kept on increasing until 4 weeks. The four cultivars showed the different patterns of shoot growth in bioreactor. The PM medium with a quarter salt strength was effective for the regeneration of axillary buds as well as shoot elongation of Jasim and Chubaek. Daese showed vigorous regeneration of axillary buds in the PM medium with a half salt strength. On the other hand, Haryeng showed slower growth than the other three cultivars.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.33-40
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• 2002

We, Tong Yang Moolsan Co. Ltd. (TYM) set up the mass-production system for virus-free seed garlic via tissue culture technique. TYM's tissue culture technique is called as 'Multiple shoot propagation technique'. This technique can lead mass propagation of genetically homogeneous seed garlic in a short period because of its highly proliferation ,ate of in vitro shoots $(15^{10}/year)$. TYM researchers applied the technique to some selected garlic cultivars with superior characteristics and carried out Held test of productivity in the inside and outside of the country for several years. According to the yearly results of Held test with virus-free seed garlic, we ascertained that virus-free seed garlic can produce the highly yield increase (max. above 50%) and also can enhance the product quality. Consequently, we estimated that TYM's seed garlic will contribute to farmers with increase of income and can elevate the national position of garlic market in the world for its competitive power of technical and production cost.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.171-177
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• 2002

We, Tong Yang Moolsan Co. Ltd. (TYM) set up the mass-production system for virus-free seed garlic via tissue culture technique. TYM's tissue culture technique is called as 'Multiple shoot propagation technique' This technique can lead mass propagation of genetically homogeneous seed garlic in a short period because of its highly proliferation rate of in vitro shoots (15/sup 10//year). TYM researchers applied the technique to some selected garlic cultivars with superior characteristics and carried out field test of productivity in the inside and outside of the country for several years. According to the yearly results of field test with virus-free seed garlic, we ascertained that virus-free seed garlic can produce the highly yield increase (max. above 50%) and also can enhance the product quality. Consequently, we estimated that TYM's seed garlic will contribute to farmers with increase of income and can elevate the national position of garlic market in the world for its competitive power of technical and production cost.

• Journal of Korean Society of Forest Science
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• v.66 no.1
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• pp.1-7
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• 1984

This study was undertaken to find out the effects of three kinds of potting media and two sources of explants on the survival and early growth of new plantlets of Poputus alba ${\times}$ P. grandidentata in the greenhouse. The results obtained can be summarized as follows; 1) Among three potting media, Terralite was best for early growth and survival of plantlets. 2) Like humidifier, an intermittent misting system can be effective in keeping relative humidity high for the plantlets. 3) Survival rates over 80% could be obtained if humidity was kept high during the hardening period. 4) During hardening period, the plantlets showed the juvenile characteristics such as smaller leaves, thinner stems, and shorter internodes. 5) There were no differences on morphological characteristics between the plantlets originating from axillary buds and the plantlets originating from multiple shoots while they were growing at the greenhouse. 6) The plantlets originating from bud culture grew normally comparing to regular cuttings.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.336-339
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• 2003

Hydrocotyle maritima Honda used as medicinal plants for a hemostatic agent was investigated for in vitro regeneration. The petiole explants of H. maritima were cultured on callus induction medium containing growth regulators ($0{\sim}5\;mg/l$, NAA and $0{\sim}5\;mg/l$ 2,4-D) either in single or in combination with $0.1{\sim}2\;mg/l$ BA for 6 weeks. Although single treatments of 2,4-D or NAA resulted in callus formation, the best results were combination of $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA. $5\;mg/l$, NAA and $0.5\;mg/l$, BA, respectively. The highest number of shoot (12 shoots per callus) was achieved with $3\;mg/l$, Kinetin. Also, when pieces of embryogenic callus induced on the medium supplemented with $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA were subcultured on hormone-free medium, somatic embryos were differentiated and developed further into welldeveloped plants.

• Proceedings of the Korean Society of Crop Science Conference
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• 2017.06a
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• pp.171-171
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• 2017

Propagation by crown division in Platycodon grandiflorum is too slow for producing many plants in a short time although the plants are uniform. This study was performed to enhance the mass propagation for Platycodon grandiflorum containing yellow green petals via various medium compositions and the growth regulators. The nodes containing yellow green petals were used as materials to execute the study with a variety of MS medium concentrations. The 1/4MS medium showed the best development of adventitious root, while the 1/2MS medium exhibited the potential growth. The higher the concentration of sucrose showed the better development and growth of both shoots and adventitious roots. Many adventitious roots were developed at the controlled culture medium at pH 4.8 with a tendency of suppression with higher levels of pH. Also, the cultivated node and leaf explants with the treatments of simple and combined applications with auxin and cytokinin at the 1/4MS culture medium with adding 5% of sucrose were used to identify the influences of growth regulators. The regeneration of plantlets at the 234single application showed a good result with the addition of BA $1mg{\cdot}L^{-1}$ and the development and growth of adventitious roots appeared to be good at the addition of NAA $1mg{\cdot}L^{-1}$. For the combined applications, the regeneration of plantlets and the development of adventitious roots were prosperous at the combined applications with BA $0.1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$. The addition of IAA for the leaf explants induced a number of plantlets that showed the potential regeneration, and the highest results was obtained from the combined applications of both BA $1mg{\cdot}L^{-1}$ and IAA $2.5mg{\cdot}L^{-1}$. In addition, the development of adventitious roots showed the satisfactory results at the combined application of both BA $1mg{\cdot}L^{-1}$ and IAA $0.5mg{\cdot}L^{-1}$.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.38-48
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• 2019

Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

• Korean Journal of Plant Resources
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• v.19 no.5
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• pp.634-639
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• 2006

In order to develop a disease-resistant root stock for the growth of watermelon, an efficient regeneration system of the gourd(Lagenaria leucantha Duch.) inbred line GO701-2 via organogenesis was established in this experiment. Using proximal parts of cotyledon explant excised from germinated seedling in vitro, maximum adventitious shoot formation (39%) was achieved on MS medium where cytokinin (BA) and auxin (IAA) were added at a concentration of 3mg/L and 0.1mg/L, respectively. Roots of the elongated shoots were successfully formed on MS medium without adding any plant growth regulators. The cucumber CsGolS1 gene known as a resistance gene against biotic and abiotic stresses, was constructed into the binary vector pBI121 under the control of CaMV 35S promoter. When the gene was introduced into the genome of gourd by Agrobacterium-mediated transformation, putative transgenic plants were obtained with the transformation efficiency of approximately 20 percent.