• Korean Journal of Plant Tissue Culture
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• v.21 no.3
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• pp.145-150
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• 1994

This study was conducted to determine the effects of 3 cytokinins (BA,2iP and kinetin) and their concentrations (0, 0.1, 0.5, 1.0, and 2.0 mg/L) on multiple shoot production of Citrus spp. 'Sambokam' and 'Byungkyool' by nodal culture. Nodal explants were obtained from in vitro germinated seedlings of both cultivars. 'Sambokam' produced more multiple shoots than did 'Byungkyool' by nodal culture. Among the 3 cytokinins tested in this study BA supplemented in semi-solid MS basal medium was the most effective stimulator for multiple shoot production, and an optimal concentration was determined to be 1.0 mg/L. Shoot elongation and root formation were inhibited by increasing cytokinin concentration, regardless of cytokinin types. BA at 1.0 mg/L produced the most multiple shoots and the highest number of leaves in 'Sambokam', whereas any cytokinin and concentration studied in this experiment did not affect any scored variables such as shoot and leaf numbers, etc. in 'Byungkyool'.

• Journal of Plant Biology
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• v.29 no.4
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• pp.233-242
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• 1986

In order to pursue some physiological studies on organogenesis in ginseng tissue culture, ginseng root explants were cultured on a modified MS medium containing NAA and kinetin. The activities of peroxidase and some enzymes were investigated and their isoenzyme patterns were also observed. The activity of peroxidase decreased by 20% in one week's culture and increased thereafter by 80% in culturing for 7 weeks compared with the control group. Glucose-6-phosphate dehydrogenase activity increased by 400% after culturing for 5 weeks and increased during the days preceeding root formation. The activities of glutamate dehydrogenase and acid phosphatase also increased during the culture. After 3 weeks' culture, new peroxidase isozyme (pH 7.6) appeared and 7 weeks' culture, another new peroxidase isozyme (pH unidentified) appeared. These patterns were also identified by using FPLC. After 7 weeks' culture, a new esterase isozyme of pH 8.5 appeared and isozyme patterns of acid phosphatase were quite changed compared with the isozyme patterns of tissue cultured for 5 weeks. In so far as these new isoenzymes appear distinctively after 7 weeks' culture, root differentiation is supposed to be induced after 7 weeks' culture.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.95-99
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• 1995

We have previously established a system for plant regeneration through somatic embryogenesis and Agrobacterium-mediated transformation of Korean ginseng. In this study to produce a fungus-resistant plant, we introduced a bean chitinase gene into ginseng using the transformation system. A binary vector pChi/748 was constructed by introducing the bean basic chitinase gene into EcoRI site of pGA748 which carries the CaMV 35S promoter governing the introduced gene and neomycin phosphotransferase II(NPT-II)gene as a positive selection marker. Cotyledonary explants were cocultured with A. tumefaciens strain LBA4404 harboring the binary vertor pChi/748 for 48 h, and transferred to MS medium supplemented with l mg/L2,4-D,0.1mg/L kinetin, 100 mg/L kanamycin, and 500mg/L carbenicillin. Kanamycin-resistant calli were formed on the cut surface of cotyledonary explants after one month of culture, and subsequently they gave rise to somatic embryos. Upon transfer onto medium containing 1 mg/L each of BA and GA$_3$, most of them converted to plantlets after 5 weeks of culture. The genomic DNA of eight kanamycin-resistant regenerants was subjected to polymerase chain reaction (PCR) using two specific 21-mer oligonucleotides derived from the chitinase gene. PCR-Southern blot analysis confirmed that the chitinase gene was incorporated into six out of the eight regenerants..

• Plant Resources
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• v.5 no.1
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• pp.29-44
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• 2002

In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.

• Korean Journal of Plant Resources
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• v.27 no.1
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• pp.43-50
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• 2014

This study was carried out to determine the optimal medium composition and growth regulators for the micropropagation of Platycodon grandiflorum (Jacq.) A. DC. Nodes containing yellow green petals were used as plant materials to execute the study. The best performance of adventitious root development was found in 1/4 strength of MS basal salt and the growth was satisfactory in the concentration of 1/2 MS medium. The best condition for adventitious root development and growth was observed in the higher concentration (5%) of sucrose and activated charcoal free 1/4MS medium respectively. Adventitious roots were developed at the controlled culture medium at pH 4.8 with a tendency of suppression with higher levels of pH. However, it was prevailed that the development and growth depended on the concentration of agar. The lower concentration of agar (0.4%) was performed better than that of higher concentration (1.2%), whereas the agar concentration (0.4%) showed the best performance for the development and growth of adventitious roots. For the development of shoots containing node, BA combined with IAA was more effective than kinetin with IAA or NAA. The highest shoot development (3.9 shoots per explant) was performed on MS medium supplemented with 0.1 mg/L BA and 0.5 mg/L IAA.

• KSBB Journal
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• v.6 no.3
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• pp.289-297
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• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• Journal of Life Science
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• v.25 no.4
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• pp.390-396
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• 2015

This study was carried out to investigate the action of phytohormones which influence the adventitious root formation of calli originating from the leaves of Persicaria perfoliata. The optimal medium condition for callus formation was ½-strength MS, 1% sucrose, and 4.5 μM 2,4-D. In order to determine which phytohormones had an effect on the adventitious root formation, the calluses were cultured in various media with different kinds of phytohormones. As a result, the medium with GA₃ or IAA was shown to induce root formation. To deeply investigate the effects of GA₃ and IAA, calli were cultured in 0.1, 1, and 10 mg/l levels of phytohormones. Numbers of roots formed per callus were 10.9, 14.2, 22.6 in GA₃, 5.8, 3.9, 1.1 in IAA, respectively. Therefore, the higher GA₃ or the lower IAA concentration, the more roots formed. To confirm this role of GA₃ we tested with inhibitors PBZ and NPA. GA₃ with PBZ resulted in reduction by 52.4~69.4% compared to GA₃ alone. In contrast, GA₃ with NPA resulted in an increase by -8~45.6% compared to GA₃ alone in root formation. Also, results were determined on the effect of GA₃ with other phytohormones on root formation. Kinetin, 2iP and ABA with GA₃ had a negative effect, but IAA with GA₃ showed a similar result to GA₃ alone. From these results we infer GA plays a key role and auxin has subsidiary activity on adventitious root formation. This is the first report that indicates GA₃ promotes adventitious root formation from calli in P. perfoliata.

• Horticultural Science & Technology
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• v.16 no.2
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• pp.239-241
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• 1998

The experiments were carried out to improve culture efficiency of rhizome and mericlone propagation through settlement of problems occurring during culture period of temperate Cymbidium species. Shooting efficiency from rhizome of C. forrestii 'Nokwoon' was improved, when cultured in $H_3P_4$ medium (Hyponex 3+peptone 4g/L) supplemented with 170mg/L $NaH_2PO_4{\cdot}H_2O$ and 0.4mg/L Thiamin e HCl, but the other varieties were not influenced to shooting efficiency by additives. Medium in which rhizome of C. nishiuchianum 'Hodukjiwha' was cultured became less browned in $H_3P_4$ medium added with 150mg/L PVP, but the other treatments of antioxidants was failed to prevent the medium browning. Re-formation of rhizome from young shoots of C. forrestii 'Sojub', 5.5cm in length occured in $H_3P_4$ enriched with 2.0 mg/L NAA and 1.0 mg/L BA under darkness, but axillary buds were elongated in the medium with 1.0 mg/L NAA and 3.0 mg/L BA under light condition. On the other hand, rhizomes from young shoot of C. forrestii 'Seosinmae' and 'Songmae', 5.5cm and 2.5cm in length respectively were reformed in 2.0 mg/L NAA and 5.0mg/L kinetin under darkness, but multishoot from young shoot were emerged in 2.0mg/L NAA and 3.0mg/L BA.

• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.247-257
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• 2005

An efficient, rapid and large-scale in vitro clonal propagation of agronomically important Indian cereal crop genotypes (NSH27 & K5) of Sorghum bicolor (L.) Moench. by enhanced shoot proliferation in shoot tip segments was designed. MS medium fortified with plant growth regulators and coconut water markedly influenced in vitro propagation of Sorghum bicolor. In vitro plantlet production system has been investigated on Murashige and Skoog (MS) medium with the synergistic combination of 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), 5% coconut water and 3% sucrose which promoted the maximum number of shoots as well as beneficial shoot length. Subculturing of shoot tip segments on a similar medium enabled continuous production of more than 100 healthy shoots with similar frequency. When the healthy shoot clumps were cultured on MS medium fortified with 6-benzyladenine ($22.2\;{\mu}M$), kinetin ($4.6\;{\mu}M$), adenine sulphate ($2.8\;{\mu}M$), ${\alpha}$-naphthaleneacetic acid ($2.7\;{\mu}M$), ascorbic acid ($30.0\;{\mu}M$) and 5% coconut water, a rapid production of axillary and adventitious buds was developed after 8 wk culture. More than 300 shoots were produced 10 wk after culture. Rooting was highest (100%) on half strength MS medium containing 22.8 mM IAA. Micropropagated plants established in garden soil, farmyard soil and sand (2:1:1) were uniform and identical to the donor plant with respect to growth characteristics. These plants grew normally without showing any traits.

• Journal of Korean Society of Forest Science
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• v.74 no.1
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• pp.56-60
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• 1986

Excised mature embryos of P. koraiensis were plated on a series of G.D. basic media supplemented with growth regulators as follow; $0.1mg/{\ell}$ NAA(M-1); $0.1mg/{\ell}$ NAA and 2,4-D (M-2); $0.1mg/{\ell}$ 2,4-D and BAP (M-3); $0.1mg/{\ell}$ 2,4-D and kinetin (M-4). Cytological study of the callus showed that thercentage of occurrance of diploid cells was observed 52% in M-3 and 36% in M-4, while that was revealed 29% in M-1 and 17% in M-1. The frequency of diploid cells was increased on the M-3 and M-4 media. The stable chromosome state is a crucial factor for organogenesis. Therefore, it can be inferred that the callus tissues cultured on media supplemented with both auxin and cytokinin have the greatest possibility of organogenesis.