• Journal of Plant Biotechnology
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• v.49 no.2
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• pp.124-130
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• 2022

The application of traditional taro propagation methods for large-scale cultivation would be insufficient to meet the high demand for quality planting materials. Therefore, this study aimed to develop an in vitro micro-propagation technique for two local taro cultivars (cv.), Wangi and Putih. Taro cormels were collected from the Malaysian Agricultural Research and Development Institute (MARDI) germplasm (Serdang, Malaysia). Explants were taken from the shoot tip of cormels and initially cultured on Murashige and Skoog (MS) basal media for four weeks. The explants were then transferred to different multiplication media supplemented with different types and concentrations of cytokinins such as 6-benzylaminopurine (BAP ) and Thidiazuron (TDZ). Shoot production was quantified after six weeks of culture. The highest mean number of new shoots was produced by the Wangi cultivar on MS medium supplemented with 2.0 mg/l BAP (2.10 shoots), MS medium supplemented with 0.5 mg/l TDZ (2.18 shoots), and Gamborg B5 medium supplemented with 6.0 mg/l BAP (2.43 shoots). The maximum average number of the Putih cultivar shoots was obtained on MS supplemented with 2.0 mg/l BAP (3.57 shoots). MS basal media was used for root initiation, as it produced an average of 25 roots with an 11-cm length. Various types of substrate mixtures were used during acclimatization. The best acclimatization substrate for the Wangi cultivar was 100% peat soil, whereas the Putih cultivar grew optimally in a combination of peat and perlites at a 1:1 ratio. Taro plantlets require approximately 4 to 6 weeks to acclimatize before they can be transferred to the field.

• Journal of Korean Society of Forest Science
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• v.79 no.2
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• pp.109-114
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• 1990

The axillary buds of 15-year-old Tilia amurensis were cultured on Saito and Ide (IS), Murashige and Skoog (MS) media and woody plant medium (WPM) to establish an effective micropropagation method. Five levels of 6-benzylaminopurine (BAP) were tested. On IS medium and WPM addition of 1.0/l BAP enhanced shoot development and shoot elongation, whereas addition of 0.5/l BAP was effective on MS medium. A better results were obtained from WPM with 1.0/l BAP and MS with 0.1/l BAP. Developed shoots were subcultured on each basal media but with 0.2/l BAP, Multiple shoots were almost doubled in a month. Root formation could be enhanced at higher concentration of indole-3-butyric acid (IBA). Better rooting rate (83.3%) was achieved on a half-strength MS medium with 3.0 /l IBA. Regenerated plantlets were successfully transferred to soil. To investigate the clonal variation in shoot development and shoot elongation by axillary bud culturing, seven plus tree clones were tested, Clonal variation in tissue culturability among plus trees was recognized by the Duncan's multiple range test at the 5% level. Kang Won No. 12 showed the best response on WPM with 1.0/l BAP.

• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.171-179
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• 2004

Hormonal requirements inducing different regeneration pathways with particular emphasis on somatic embryo-genesis in Alhagi graecorum were studied. While combination of 0.5 $\mu{M}$ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.5 $\mu{M}$ 6-benzylaminopurine (BAP) and 5 $\mu{M}$ 1-naphthaleneacetic acid (NAA) in MS medium induced callus formation and callus maintenance from internodal explants, each alone or in combination with other induced distinct regeneration pathway. Adventitious bud formation was induced on MS medium supplemented with 2.5 $\mu{M}$ BAP. It was improved when 2.5 $\mu{M}$ BAP was used in combination with 5 $\mu{M}$ NAA. MS medium containing 0.5 $\mu{M}$ 2,4-D or 5 $\mu{M}$ NAA induced the formation of abnormal direct somatic embryos. While increase of 2,4-D concentration (1.125-9) resulted in the formation of viable embryogenic mass, increase of NAA did not change its effect. NAA should be used in combination with 2,4-D even at low concentration (0.5 $\mu{M}$) to form embryogenic mass. In A. gaecorum, the role of 2,4-D as trigger of somatic embryogenesis and BAP as trigger of adventitious bud formation was deduced, but for maximum yield certain auxin-cytokinin ratio should be applied. Embryogenic masses characterized by high water content, low peroxidase activity, and low number of peroxidase and glutamate oxaloacetate transaminase bands in comparison with calli obtained under conditions stimulating adventitious bud formation. The resulted differential gene expression, which could be detected by native-PAGE patterns, could be used as marker for organogenic pathway in A. graecorum.

• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.241-246
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• 2005

Factors affecting regeneration of different Rubus varieties (blackberry, raspberry and their hybrid) were examined and a reliable regeneration system was established. Media for stock plant maintenance were tested; different explants and media were investigated to find the best circumstances for the regeneration. The effect of the commonly used antibiotics was studied to determine the most suitable one for selection of the transformants. We found that both MS and LS media supplemented by $20\;gL^{-1}$ sucrose are suitable for the stock plant maintenance. The optimal hormone content for the stock plants is $0.125\;mgL^{-1}$ 6-benzylaminopurine (BAP) with $0.01\;mgL^{-1}$ indole-3- butyric acid (IBA). The highest regeneration rate was observed on medium containing MS salts with B5 vitamins complemented with glucose, sucrose, maltose, $10\;gL^{-1}$ each, supplemented with benzylaminopurine riboside (BAR) ($2\;mgL^{-1}$) and indole-3-acetic acid (IAA) ($0.1\;mgL^{-1}$). The regenerated shoots appeared directly from the cut edges, without callus phase. Hygromycin and geneticin proved to be good selection agents for the Rubus explants, but due to their severe effect on the tissues we propose to use marker-free constructions for the transformation.

• BMB Reports
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• v.32 no.1
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• pp.51-59
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• 1999

The Korean radish cationic peroxidase (KRCP) promoter, comprising nucleotides -471 to +704 relative to the transcriptional initiation site, was fused to the GUS gene and transformed to tobacco BY-2 cells. We examined how auxin (2,4-dichlorophenoxyacetic acid, 2,4-D), cytokinin (6-benzylaminopurine, BAP), gibberellic acid ($GA_3$), abscisic acid (ABA), methyl jasmonate (MeJA), and phosphatidic acid (PA) affect the GUS expression in the presence or absence of 2,4-D in a modified LS medium. Exogenous 2,4-D or BAP greatly decreased the GUS expression regulated by the KRCP promoter in a modified LS medium containing 0.2 mg/l 2,4-D. $GA_3$ increased the GUS expression and ABA completely reduced the inductive effect of $GA_3$. The GUS expression was also increased dose-dependently by plant defense regulators, MeJA and PA. In contrast to the above results, auxin deprivation from the modified LS medium increased the GUS expression after treatment with exogenous 2,4-D whereas BAP still greatly decreased the GUS expression dose-dependently. $GA_3$ or MeJA slightly decreased the GUS expression. The data suggest that auxin deprivation changes the sensitivity of the suspension cells to exogenous chemicals and that the regulation of the KRCP promoter by 2,4-D, $GA_3$, and MeJA is dependent on auxin, whereas the regulation by BAP is not. This study will be valuable for understanding the function and expression mode of the Korean radish cationic peroxidase in Korean radish.

• Korean Journal of Medicinal Crop Science
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• v.11 no.4
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• pp.316-320
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• 2003

A protocol is described for rapid multiplication of Zanthoxylum piperitum DC. (Rutaceae), an important aromatic and medicinal plant, through shoot-tip explant cultures. Murashige and Skoog (MS) medium supplemented with various concentrations of N-6-benzyladenine (BA), N-6-benzylaminopurine (BAP) and thidiazuron (TDZ), in single or in combination with ${\alpha}-naphthaleneacetic$ acid (NAA), was used to determine the rate of shoot proliferation. N-6-benzyladenine (BA) used at 0.5mg/l, was the most effective in initiating multiple shoot proliferation at the rate of 23 microshoots per shoot-tip explants after 40 days of culture. Shoot multiplication increased 1.2-fold in each successive subculture. Induction of rooting (98%) was achieved by transferring the shoots to the same basal medium containing 2 mg/l indole-3-butyric acid (IBA). Plantlets went through a hardening phase in a controlled growth chamber, prior to in vivo transfer. These results represented that possible application for the mass production of plantlets through in vitro culture system of Zanthoxylum piperitum DC.

• Korean Journal of Plant Resources
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• v.24 no.5
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• pp.628-635
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• 2011

This study was conducted to establish the tissue culture system for Korean domestic Miscanthus sinensis, which is used in various purposes such as forage, and bio-energy resources. With the mature seed of Miscanthus, optimum concentrations of plant growth regulators were identified for an efficient callus induction and regeneration. Among the treatments of 1~10 $mg{\cdot}L^{-1}$ 2,4-D, IBA, or NAA, callus induction rate was highest (85.3%) on MS medium containing 5 $mg{\cdot}L^{-1}$ 2,4-D. Under the condition, the callus were efficiently induced and proliferated with comparably lower frequencies of callus browning. In shoot regeneration, the treatment of NAA combined with BAP seemed to contribute more efficient conditions to shoot regeneration than those of NAA with Kinetin or 2-iP. Especially, regeneration efficiency and number of regenerated plants were 83.7% and 5.5 in 3 $mg{\cdot}L^{-1}$ NAA with 5 $mg{\cdot}L^{-1}$ BAP, respectively, which were higher frequencies than those in NAA with Kinetin or 2-iP. In results, 5 $mg{\cdot}L^{-1}$ 2,4-D and 3 $mg{\cdot}L^{-1}$ NAA combined with 5 $mg{\cdot}L^{-1}$ BAP were efficient for embryogenic callus induction and regeneration of Miscanthus. This system would be useful for mass-propagation and developing new cultivars via tissue culture of Miscanthus sinensis.

• Journal of Plant Biology
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• v.28 no.3
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• pp.233-241
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• 1985

The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl2.2H2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 $\mu$M $\alpha$-naphthaleneacetic acid (NAA) and 32 $\mu$M N6-benzylaminopurine (BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.

• Plant Resources
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• v.4 no.1
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• pp.53-58
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• 2001

The effect of carbenicillin on the dedifferentiation and the regeneration efficiency of plant tissues of horseradish(Armoracia rusticana) was evaluated, Inhibition effect for callus initiation was observed when leaf blade, root and petiole segments were grown on MS medium containing 500 mg/L to 2000 mg/L carbenicillin and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The regeneration of horseradish shoots from leaf blade, root and petiole explants were decreased as the addition of carbenicillin increased from 1000 mg/L to 2000 mg/L in MS medium containing 0.5 mg/L of 6-benzylaminopurine (BAP) or kinetin. Especially, 500 mg/L carbenicillin treatment significantly inhibited shoot induction when leaf blade explants were grown on hormone-free MS medium. It was suggested that the toxic effects of combinations of carbenicillin and 2,4-D may be due to high auxin activity levels.

• Journal of Plant Biotechnology
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• v.39 no.1
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• pp.69-74
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• 2012

Using calli of $Muscari$ $armeniacum$ cv. 'Early Giant' that is monocotyledonous ornamental bulb crop with increasing demand in Korea, we carried out current studies to establish an in vitro multiple propagation protocol via somatic embryogenesis. We found that soft pale yellow green calli were induced from leaf explants cultured on all media containing 0.1~3.0 $mg{\cdot}L^{-1}$ auxins such as 1-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). However, induced calli showed vigorous growth only when they further transferred on same media containing 2,4-D, 4-amino-3,5,6-tri-chloropicolinic acid (picloram), or 3,6-dichloro-o-anisic acid (dicamba). Although frequency of somatic embryo induction depended on callus source and PGR composition in somatic embryo induction media, somatic embryogenesis was initiated on surface of proliferated calli after transferring on media with no PGR or 0.01 $mg{\cdot}L^{-1}$ NAA co-supplemented with various cytokinins such as $N^6$-benzylaminopurine (BAP). Highest number of embryo at 9.3 per callus clump was obtained when calli which were grown under 0.1 $mg{\cdot}L^{-1}$ picloram supplementation were sub-cultured on medium with 0.01 $mg{\cdot}L^{-1}$ NAA and 0.5 $mg{\cdot}L^{-1}$ BAP. In addition, morphological characteristics of somatic embryo were categorized into following nine phases: globular, biased heart, biased torpedo, early cotyledonary, middle cotyledonary, late cotyledonary, early sprouting, middle sprouting, and late sprouting embryos.