• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.97-102
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• 2004

This study was carried out to establish The regeneration of healthy seedlings from the nodal segment culture of Chinese yam (Dioscorea opposita cv. Danma), cultivated in Korea. Different explants such as leaves, petioles, roots and nodal pieces, excised from the in vitro grown seedlings of Chinese yam, were cultured on MS medium supplemented with various combinations of growth regulators. All the growth regulators used induced plantlet regeneration from the nodal segments at a high frequency, while there was no induction of shoot or callus from leaf, petiole or root tissues. The medium supplemented with 0.01mg/L NAA, 0.5mg/L BA, 0.5-1.0mg/L kinetin and without plant growth regulator was effective for shoot development of buds from the nodal segment culture. The concentration of BA and NAA was an important factor in the bud induction of buds from the nodal segments of Chinese yam. Nodal segments cultured on the medium containing 1.0mg/L NAA and 0.5-1.0mg/L BA gave the best response to bud formation. The addition of GA$_3$ to the culture medium suppressed shoot induction and growth, while it increased microtuber formation. The shoot growth and microtuber formation were also affected by medium strength and solidity. The MS basal medium containing 1 g/L gelrite was suitable for microtuber formation from the nodal segment of Chinese yam.

• Korean Journal of Plant Resources
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• v.19 no.2
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• pp.308-314
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• 2006

This experiment was conducted out to investigate the effect of plant growth regulators on callus and shoot formation. The callus formation was effective on 1/2 MS medium containing 2,4-D, while shoot formation was suppressed. Shoot formation and differentiation were the highest in combination 0.1 mg/L of IAA and 0.1 mg/L of BA. The polarity of explants was investigated from cotyledon, which excised 20% of each basal and terminal parts. Formation of shoot was induced from excised ends of the basal part. In excised ends of the basal part, callus was induced vigorously and shoots were produced lately. Root induction was easily achieved in 1/3 MS medium from the adventitious shoot and more than 90% of regenerated plantlets acclimatized successfully and flowered normally.

• Journal of Plant Biotechnology
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• v.45 no.1
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• pp.55-62
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• 2018

In the present study in vitro mutagenesis was used to study the effect of gamma irradiation and EMS on callus induction, morphogenesis and production of multiple shoots from different explants of Citrullus colocynthis (L.) Schrad. Gamma radiations (5 kR to 20 kR) and certain chemicals have been effected on plant growth developments and changes of biochemical metabolisms in plants. Murashige and Skoog (MS) medium containing with auxins such as NAA, IAA, 2,4-D (0.5 ~ 2.0 mg/l), cytokinines BAP, kn TDZ, (0.5 ~ 2.5 mg/l), L-Glutamic acid (1 ~ 2 mg/l) and Coconut milk (10 ~ 20%). After 5 weeks on induction media, explants and callus (EC) were exposed to 5 kR, 10 kR, 15 kR and 20kR, of gamma radiation and treated with 1, 2, 3, 4 and 5 mM ethyl methane sulphonate (EMS) for 30 min. The highest percentage of callusing was observed (70%) stem irradiated with 5 kR and significantly decrease in fresh and dry weight of callus in the below 4 kR doses and above 20 kR doses, there was a progressive decrease in the fresh weight and dry weights when compared to control callus. Maximum percentage of plantlet regeneration (59%) was induced from callus exposed to 15 kR gamma irradiation on MS media fortified with 2.0 mg/l 2,4-D + 2.0 mg/l BAP + 2.0 mg/l L-glutamic acid. Increase in gamma irradiation dose above 15 kR and 5 mM EMS reduced regeneration capacity of callus. Doses higher than 20 kR and 7 mM EMS was lethal to micropropagated plants of Citurullus colocynthis.

• Journal of Korean Society of Forest Science
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• v.97 no.1
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• pp.127-133
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• 2008

This study was conducted to establish the optimal condition for plant regeneration and acclimatization from somatic embryos of Panax ginseng. Cotyledon segments of Panax ginseng produced primary and secondary somatic embryos when cultured on MS and WPM media supplemented with 7% sucrose. To induce plantlet conversion, cotyledonary somatic embryos were cultured on WPM solid medium with $GA_3$ at various concentrations (1~30 mg/L) for 4 weeks. Plantlets were transferred to 1/2 WPM solid medium with $GA_3$ at various concentrations (0~5 mg/L) and 0.5% activated charcoal for shoot and root elongations. Elongated plantlets further developed into well-developed leaf and root system on 1/3 SH medium with 0.5% activated charcoal under ventilation condition for 5 months. The highest survival rate to soil was 75% when plantlets were regenerated on 1/3 SH medium without sucrose under ventilation condition.

• Plant Biotechnology Reports
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• v.4 no.4
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• pp.261-267
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• 2010

We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

• Korean Journal of Medicinal Crop Science
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• v.20 no.1
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• pp.47-53
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• 2012

This study was conducted to examine effects of plant growth regulators and light resources on the formation of multiple shoot and plant regeneration of Hovenia dulcis var. koreana Nakai. Stem and shoot tip were cultured on MS medium or WPM supplemented with various plant growth regulators. At the single treatment, the highest shoot formation was obtained when stem explants were cultured on WPM supplemented with kinetin $1.0mg{\cdot}L^{-1}$. MS medium containing NAA 0.1 and TDZ $0.1mg{\cdot}L^{-1}$ gave the best results for shoot induction rate and shoot growth in combination treatments. Of the BAP and kinetin tested, BAP $0.5mg{\cdot}L^{-1}$ on WPM was found to be more effective for shoot growth from shoot tip. Under white fluorescent light treatment, shoot growth was much higher than blue, red LED treatments. Root induction from in vitro growth of plantlet was the best on WPM supplemented with $1.0mg{\cdot}L^{-1}$ IBA. The results suggest that selection of plant growth regulators and light resources could be important factor to achieve an efficient in vitro growth.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.171-175
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• 1998

Chinese foxglove (Rehmannia glutinosa) is receiving much attention as one of the principal medicinal crops and the crude drug damand expands rapidly.This study was conducted to obtain the basic breeding information of Chinese foxglove. Effects of supplemental plant growth regulators were investigated on leaf tissue for proliferation. 100% callus formation, 31% plantlet regeneration and 6% root differentiation were obtained by adding 0.5 mg/L NAA and 2.0 mg/L BA. 2,4-D and Zeatin treatment also resulted in 95% increase in callus formation, but shoot was not formed. During the subculture, callus propagation rate recorded 15.4% with 0.2 mg/L NAA and 1.0 mg/L BA and plant regeneration improved on MS medium supplemented with 0.2 mg/L NAA and 0.5 mg/L kinetin. The number of shoot formed ranged from 1.7 on WPM medium to 3.4 on MS medium with 0.1 mg/L NAA and 0.5 mg/L BA. Supplementation of 1.0 g/L activated charcoal improved the In vitro plant growth.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.81-86
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• 2008

This study describes conditions for plant regeneration from protoplasts-derived from embryogenic callus of satsuma mandarin. Plants were generated via somatic embryogenesis. Protoplasts isolated directly from nucellar callus induced from immature ovule of satsuma mandarin cv. Okitsu (Citrus unshiu Marc.) were cultured in 0.6M $BH_3$ medium. Cell division and plating efficiency were affected by protoplast culture method. The liquid over solid method was the most effective for formation of microcalli. Most of microcalli grew rapidly and transferred onto embryoid formation medium. Optimum embryoid formation medium was MT medium containing 1.5 g/L malt extract, 0.146 M sucrose and the medium for plantlet regeneration was MS medium containing 0.09M sucrose, 1.0 mg/L $GA_3$. No differences were noticed in growth habits and leaf characters such as shape, thickness, and colour between protoplast-derived plants and nucellar seedlings. This plant regeneration system from protoplasts-derived from embryogenic callus provides an alternative way for producing new scion and rootstock cultivar from citrus species which can not be crossed.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.23-28
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• 1994

This study was rimed out to examine the difference in the cell vitality between mesophyII protoplast (MP) and paraveinal mesophyII protoplast (PVMP) of Nicotiana tabaccum 'Xanti', Petunia hybrida 'Blue Star' and Chrysanthemum morifolium 'Baeckwang' by using urea permeability technique. The effects of various enzyme solutions and incubation time, NAA and thidiazron on plant regeneration from isolated protoplasts were also investigated. The vibratome technique was used for protoplast isolation and urea permeability test because the fresh living, thin tissue stripes (50 ${\mu}{\textrm}{m}$ of thickness) could be obtained with minimal damage with the vibratome. For the three plants examined, the urea permeability on the tested tissue stripes was relatively higher in PVMP than in MP by about Ks = 2.0 $\times$ 10$^{-5}$ cm/sec. The treatment of an enzyme mixture of 1.5% cellulase R-10, 1% Driselase, 0.5% Macerozyme R-10, and 0.5% Pectinase for 4 to 8 h was effective on the isolation of PVMP. The highest frequency of callus formation and plant regeneration from the isolated protoplasts was obtained with NAA 2 mg/L and thidiazuron 0.01 mg/L. Furthermore, the results demonstrated that cell devision and plantlet regeneration was more frequent in the PVMP than in the MP of the same leaf or plant We, therefore, conclude that UM is an excellent experimental material for the callus formation and regeneration from isolated protoplasts.

• Korean Journal of Plant Resources
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• v.12 no.2
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• pp.107-119
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• 1999

In this study, the induction regeneration of callus from immature embryo in trifoliata orange (Poncirus trifoliata RAFIN.) were accomplished. The embryogenic calli were induced from the immature embryo derived from seed when the calli were irradiated for 16hr at about 2,000 Lux in $\frac{1}{2}$ MS medium supplemented with 3% sucrose, and 44.4$\mu$M BA. Regeneration to whole plants was the most successful in MS medium containing 5.0$\mu$M BA. The yellowish callus was developed at 2 to 3 weeks of culture and the callus was changed from yellow to green at 5 to 6 weeks culture. In vitro regeneration was directly induced from embryogenic callus in MS medium containing 3% sucrose and 5.0$\mu$M BA. Multishoot was formed at 16 weeks culture. Moreover, when the root-formed plantlet was transplanted to soil, they grew to a whole plant. The compact cultured-cells were observed by light microscope after 4 weeks of cultivation and the embryogenic clumps were formed about the 5 weeks. At the same time, the neighboring cells were liquefied. In addition, differentiation of leaf and stem from the callus was observed after 12 weeks. The developed oil sacs and the profacicular cambium of the immature leaf were observed after 18 weeks. Therefore, we can see the considerable changes of cell arrangements according to the developmental stages of calli from trifoliata orange.