• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.88-94
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• 2005

Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

• Journal of Korean Society of Forest Science
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• v.97 no.4
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• pp.452-457
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• 2008

In order to develop an efficient micropropagation technique effect of plant growth regulators (PGRs) affecting on shoot proliferation from shoot apex in Pyrus ussuriensis was tested. Generally, there was no conspicuous effect on shoot induction by the treatment of PGRs and one or two shoots/explant were induced when cultured on MS medium supplemented with BA and/or BA plus NAA. Both apical shoot necrosis and hyperhydric shoots were observed frequently in multiplied shoots, and callus was formed at the basal part of shoots. About 20% spontaneous rooting was achieved in growing shoots, however the proliferated shoots exhibited poor rooting rate in gelrite supported media. When we tried to ex vitro rooting of the shoot cutting, the shoot cuttings rooted up to 50% with 100 mg/L IBA application. The rooted plantlets grew normally after acclimatization in the greenhouse.

• Korean Journal of Medicinal Crop Science
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• v.21 no.2
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• pp.142-147
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• 2013

Gastrodia elata has been cultivated as an important medicinal resources to treat various human diseases. One of the major problems associated with its field production is the degeneration of seed tubers, which is mainly caused by soil-borne pathogens. This study was conducted to produce disease-free seed tubers by the development of in vitro micropropagation method. First, tubers of G. elata were treated with $HgCl_2$ prior to culturing in vitro. Among various culture medium tested, water agar (WA) and WPM medium were the most effective on the induction and growth of vegetative stems. NAA ($0.1mg/{\ell}$) or TDZ ($1.0mg/{\ell}$) in WA medium showed better growth of vegetative stems compared to other plant hormones. Finally the induction and growth of vegetative stems were better in the dark compared to the light condition. In this study, we established an in vitro micropropagation system of G. elata, which might be an efficient way to increase the yield and quality of G. elata tubers in the field production.

• Journal of Crop Science and Biotechnology
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• v.10 no.3
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• pp.141-146
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• 2007

The roots of Aristolochia elegans Mast.(Aristolochiaceae) are widely used in Mexican traditional medicine as a remedy for scorpion venom. Current experimental evidence supports its purported antidote properties. However, collection from the wilderness has lead to local extinction of natural populations. In order to contribute to species preservation, cultivation, and standardization of morphological and pharmacological properties, a micropropagation method was developed. This includes in-vitro germination of seeds to produce aseptic plantlets, induction of multiple budding, and acclimatization. The treatment with benzylamino purine(10 ${\mu}M$) induced the highest number of buds(3.1 on average) in both types of explants. On the other hand, indolebutyric acid(1.5 ${\mu}M$) caused the highest root index(11.8) per explant. One hundred percent of the micropropagated plantlets developed vigorously after the acclimatization process.

• Korean Journal of Plant Tissue Culture
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• v.22 no.4
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• pp.223-227
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• 1995

Immature flowers and lateral buds of Neoregeria carorinae cv. Tricolor were cultured for micropropagation and the collecting times of materials, growth regulators and theirs concentrations, and cultural methods on the formation of adventitious buds and growth were investigated in this experiment The formation rate was the highest in immature flowers collected at 4weeks after flower bud differentiation and in buds at 7weeks after flower differentiation of adventitious buds. MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L BA was the most favorable for the formation of adventitious buds. Solid medium was more effective for the formation of adventitious buds than liquid one. MS medium with 1.0 mg/L NAA was the most suitable for the rooting of regenerated shoot. Liquid medium was effective for the rooting of regenerated shoot than solid one.

• Journal of Plant Biotechnology
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• v.41 no.2
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• pp.100-106
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• 2014

In order to develop an efficient in vitro micropropagation technique for a rare plant species, Astragalus membranaceus Bunge var. alpinus N., shoot proliferation and in vitro or in vivo rootings were conducted and hyperhydrated leaf generated from cultures was histologically observed. During shoot induction, no distinct effect on multiple shoot induction was found between BA and kinetin treatment. BA enhanced the number of internodes, whereas kinetin stimulated shoot elongation. Hyperhydrated leaf composed of bigger cells and retarded palisade parenchyma and showed irregular cell arrangement compared to normal leaf. Especially starch content in hyperhydrated leaf was significantly reduced. The best rooting rate was achieved by B5 medium among three different medium (B5, MS and WPM) and 0.1mg/L IBA treatment induced the highest rooting ratio (80%). No statistical difference was induced by explant types (apical bud or axillary bud) in terms of rooting ratio. In vivo cutting induced rooting rate up to 65% by 0.5% IBA/Talc powder treatment. Although in vivo rooting rate was less efficient compared to in vitro rooting, better survival rate was observed after soil acclimatization. Present study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the species.

• Korean Journal of Medicinal Crop Science
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• v.2 no.2
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• pp.154-161
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• 1994

This experiment was carried out to establish micropropagation system in Fritillaria thunbergii Miq. Through the culture of bulblet scales, stems, node-buds and shoot tips with special reference to the effect of physiological age of explant and plant growth regulators on bulblet formation. Number of formed bulblets was significantly increased in node-bud or stem tissue compared to scals segments and on the medium supplemented with kinetin than BA containing medium. Optimum levels of kinetin for bulblet formation from node-bud taken from above 3 cm shoot length and stem segments excised from below 3 cm shoot length were 5.0 mg /L and $1.0{\sim}3.0\;mg$ /L kinetin, respectively. Interesting phenomenon was observed, the direct formation of bulblets from the axilliary bud of cultured explants. Bulblet forming capacity in stem tissue was depended on stem age, young stem had high regeneration ability compared to old stem taken from above 10 cm shoot length. 1.0 mg /L kinetin was optimum concentration for the formation of bulblets from old stem segments. Stem tissue taken from underground growing plant was promoted coampare to shoot tips or bulb scale segments. Optimum concentration of sucrose was $5{\sim}7%$. Summariged above results revealed that effective explant for micropropagation was stem and /or node-bud tissue excised from less than 3 cm plant height compared to those of bulb scale segments which showed high contamination after culture. Maximum multiplication rate of young stem and /or node-bud segment was about 20 times. Kinetin requirement for stimulation of bulblet formation from cultured explant depended on source of explants but favorable levels of kinetin for organogenesis ranged from 1.0 mg /L to 5.0 mg /L.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.63-69
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• 2000

In order to establish a micropropagation system for buffalo gourd (Cucurbita foetidissima ) and common milkweed (Asclepias syriaca), the effects of several plant growth regulators and culture temperature on shoot multiplication and rooting were investigated. In buffalo gourd, the greatest number of shoot from shoot tip culture and well growth of formed shoot were obtained on the MIS medium supplemented with 1.0 mg/L BA and 0.3 or 0.6 mg/L IAA. Whereas kinetin and 2iP were not effective for shoot multiplication in vitro. It was found that 22$^{\circ}C$ and $25^{\circ}C$ were suitable for shoot multiplication. Roots were easily formed by the addition of auxins, especially 1.0 or 2.0 mg/L IBA and 2.0 mg/L IAA. Over 90% of plants survived successfully after being transferred into the field. In common milkweed, BA was more effective than kinetin or 2iP for its micropropagation in vitro. The increased shoot weight and number of nodes per shoot were obtained on the medium containing 3.0 mg/L BA and 0.3 or 0.6 mg/L IAA. But 2iP promoted the shoot elongation. In addition. common milkweed was sensitive to culture temperature in vitro. Temperature around 22$^{\circ}C$ was favorable for shoot multiplication and growth, whereas temperature higher than $25^{\circ}C$ usually reduced the rate of shoot survival rate.

• Journal of Korean Society of Forest Science
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• v.84 no.1
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• pp.41-47
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• 1995

A micropropagation system for black Locust(Robinia pseudoacacia) was established by using shoots and pin-punctured leaves of in vitro germinated seedlings. The greatest number of shoots (an average of 10.5 shoots) was obtained when shoot tips were cultured on MS medium supplemented with 1.0 mg/l BAP and 0.01 mg/l NAA. When pin-punctured leaf explants were cultured on the same medium, mean number of 13.5 shoots were produced. Shoot growth was accelerated by adding 50 mg/l of silver nitrate ($AgNO_3$), an anti-ethylene compound to the culture medium. Each shoot was excised from the mass and transferred onto half strength MS medium for rooting. Zygotic embryos at different developmental stages were cultured on LS medium supplemented with various growth regulators to induce somatic embryos. When cultured on LS medium with 1.0 mg/l 2,4-D. 14.3% of the zygotic embryos induced somatic embryos. Upon transfer onto the basal medium, somatic embryos sporadically converted into plantlets.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.117-121
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• 2002

We have developed an in vitro micropropagation system via shoot formation from axillary buds using nodal segments of Corylopsis coreana. Explants from both juvenile tree (one-year-old greenhouse stock seedlings) and mature tree (ten-years-old tree in nursery) were compared with regard to propagation efficiency. Combined treatment of both BA and zeatin were effective on shoot proliferation since the best result was obtained on MS medium supplemented with 0.5∼3.0 mg/L zeatin and 0.2 mg/L BA. Generally, juvenile explants were better in both shoot proliferation and growth than mature explants. However, as the duration of in vitro culture was proceed to 6 months, explants from mature tree also produced three shoots per explant. Distinctive differences in rooting and adaptability to soil of shoots obtained from mother trees. Whereas shoots originated from juvenile explants rooted as high as 97%, those from adult explants showed 62% rooting. Similar result was also observed in soil acclimatization. The plantlets derived from juvenile plants survived 67%, while only 48% of those from adult trees survived. The results showed a possibility of the micropropagation of Corylopsis coreana through shoot formation from axillary buds. In addition, the advance of the research still remain to enhance the frequency of acclimatization of plantlets from mature trees for practical application.