• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

• FLOWER RESEARCH JOURNAL
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• v.16 no.1
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• pp.12-16
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• 2008

This study was carried out to produce haploid plants to verify a systematic breeding program and genetic analysis. Effect of developmental stage of pollen grains and pre-treatment temperature on production of haploid plants was investigated. Microscopic investigation of the explants (Lilium Asiatic hybrid 'reamland' revealed that the length of flower bud at 23.0-24.9, 25.0-26.9, and 27.0-28.9 mm long coincided with tetrad, uninucleate, and binucleate, respectively. When the efficiency of the anther culture from microgametogenetic stages was tested, late uninucleate to early binucleate stage, having the length of 23.0 to 28.0 mm long flower bud, was the best. The frequencies of the callus induction and plant regeneration from the stage mentioned above were 17.8 and 6.7%, respectively. When calli were cultured on the MS medium containing picloram and zeatin at $25^{\circ}C$, shoots were obtained. Roots of regenerated plantlets were confirmed as haploid through an microscopic observation.

• 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.

• Journal of Bio-Environment Control
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• v.26 no.2
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• pp.72-77
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• 2017

This study was conducted to examine the effect by application method and concentration of plant growth regulator (PGR) on the growth and runner production of strawberry (Fragaria ${\times}$ ananassa Duch. cv. Maehyang) in a velno-type greenhouse. The seedlings of strawberry were transplanted in pot ($64{\times}27{\times}18cm$) filled with commercial mixed medium (Tosilee) on February 22nd, 2016. The 6-benzylaminopurine (6-BAP) was applied with foliar spray or drench, respectively as 900, 1,200 or $1,500mg{\cdot}L^{-1}$ (50 mL per plant) at 3 weeks after transplanting. Nutrient solution was sufficiently supplied by the drip irrigation as EC $0.65dS{\cdot}m^{-1}$ for rooting during 7 days. After rooting, the 450 mL nutrient solution supplied per pot twice a day (10 min). Plant height and crown diameter of 'Maehyang' mother plant appeared no significantly difference. The other growth characteristics, such as root length, number of primary roots, leaf length, leaf width, leaf area and fresh and dry weights of the shoot or root, were significantly the greatest in the control. And, the SPAD value of strawberry was the highest as 44.2 in the drench with $900mg{\cdot}L^{-1}$. The foiler spray was more effective in runner production than drench, and the number of runners appeared high values at the 900 and $1,500mg{\cdot}L^{-1}$. Whereas, the number of strawberry plantlets was effective in the drench. The results indicate that both growth and the number of runners of strawberry plant were the best achieved by foliar spray application at the $900mg{\cdot}L^{-1}$.

• Journal of Bio-Environment Control
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• v.23 no.2
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• pp.144-147
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• 2014

It was intended to closely examine an effect that a change in the concentration of culture medium had on the potato(Solanum tuberosum L.) plantlet growth in the microponic system so as to mass-produce the virus-free plant of new variety 'Saebong' for potato processing. The adjusted concentration of potato culture medium was 0.2, 0.6, 1.0, 1.4, 1.8, and $14.0dS{\cdot}m^{-1}$. And potato seedling was cut into pieces of 1.5 cm in length, which included 2 growth points and leaves. And each was explanted in glass vial of 50 mL. And experiments were carried out twice for 18 days or 21days. Culture medium of 2ml was put in the container respectively. And 1 mL was added after 10 days. And in terms of cultivation environment, the experiment was carried out at the day length of 16 hours at the temperature of $23{\pm}1^{\circ}C$ under the white LED light of $40{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$. The concentration of culture medium in the experiment I was EC 0.2, 1.0, $14dS{\cdot}m^{-1}$ and was adjusted to 0.6, 1.0, 1.4, $1.8dS{\cdot}m^{-1}$ in the experiment II. The results showed that the survival rate of plantlet was 90% at $0.2dS^2m^{-1}$, 100% at $0.6dS^2m^{-1}$, 100% at $1.0dS^2m^{-1}$. 0% at $1.4dS{\cdot}m^{-1}$, 0% at $1.8dS{\cdot}m^{-1}$. and 0% at $14.0dS{\cdot}m^{-1}$ after 7 days. With regard to the explanted potato seedling, in case of the treatment where the electrical conductivity of culture medium was adjusted to $1.0dS{\cdot}m^{-1}$, root developed 2 days after transplantation. And the plantlet vigorously grew into strong plant that had 7 leaves, length of 5cm, and fresh weight of 0.5 g after 18 days. In case of the treatment where the concentration of culture medium was adjusted to $0.6dS{\cdot}m^{-1}$, the root plantlets developed 4 days after transplantation. And those grew into plant that had 7 leaves and fresh weight of 0.2 g after 21 days. Therefore, we found that it is effective to control potato culture medium by adjusting its electrical conductivity to $0.6{\sim}1.0dS{\cdot}m^{-1}$ for the mass production of virus-free potato seedling in the microponic system.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.116-122
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• 2014

Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.

• Korean Journal of Medicinal Crop Science
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• v.2 no.1
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• pp.60-66
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• 1994

This study was conducted to determine the optimum conditions of inducing callus, proliferating callus, forming somatic embryos, and regenerating plantlets via somatic embryogenesis, for the purpose of producing artificial seeds and substantially developing plant factory technologies that can be employed to all seasons production of Bupleurum plants. Callus was efficiently induced from leaf tissues at three leaf stage in the MS medium supplemented with 2, 4-D 2mg /1 and thidiazuron(TDZ) 0.lmg /1. Callus induction from leaf tissues at maturity was mostly effective in the mixture of 2,4- D 2mg /1 and TDZ 1.0mg /1 while that from flower bud tissues was fairly good in the MS medium containing 2,4-D 1 or 2mg /1.Callus was formed in 15 to 20 days after culture initiation in the MS media supplemented with 2, 4- D 1-2mg /1 and TDZ 0.l-1.0mg /1. Such hormones as kinetin 3mg /1, GA 1mg /1, and the mixture of GA 1mg /1 and TDZ 1mg /1 effected markedly to proliferate the callus cells.The optimum temperature and light intensity for callus culture were found to be $25^{\circ}C$ and 3000 Lux, respectively. Direct plant regeneration from cultured callus was fairly made on hormone-free MS or half-strength MS medium. Somatic embryogenesis was most frequently observed in hormone-free media:60 somatic embryos per 20ml in MS medium and 28 somatic embryos per 20ml in half -strength MS medium. There were three stages-globular, heart, and torpedo-in development of somatic embryos, among which globular stage was more frequently observed in MS medium rather than in half-strength MS medium. Somatic embryos induced from suspension culture fairly differentiated a number of shoots and roots on hormone-free and half-strength MS solid medium.