• Plant Biotechnology Reports
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• v.4 no.4
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• pp.321-328
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• 2010

An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro (15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g $1^{-1}$ sucrose, 2.2 g $1^{-1}$ Gelrite, and 7.7 lM naphthalene acetic acid (NAA) with 2.2 ${\mu}M$ thidiazuron (TDZ). Regeneration of adventitious shoots from callus (30-40 shoots per explant) was achieved on MS medium containing 5.5 ${\mu}M$ TDZ, 2.2 ${\mu}M$ NAA, and 3.3 ${\mu}M$ silver nitrate ($AgNO_3$). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 ${\mu}M$ gibberellic acid ($GA_3$). The elongated shoots were rooted in MS medium supplemented with 4.0 ${\mu}M$ indole 3-butyric acid (IBA). Rooted plants were acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an average of 40 plants per leaf explant with a culture period of 98 days.

• Journal of Plant Biotechnology
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• v.47 no.1
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• pp.53-65
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• 2020

Here, we describe an efficient and rapid protocol for the micropropagation of Thymus pallidus Cosson ex Batt., a very rare medicinal and aromatic plant in Morocco. After seed germination, we tested the effect of different macronutrients, cytokinins alone or in combination with gibberellic acid (GA₃) or auxins, on T. pallidus plantlet growth. We found that Margara macronutrients (N₃₀K) had the best effect on the in vitro development of the plantlets. The addition of 0.93 μM/L 1,3-diphenylurea (DPU), 0.46 μM/L adenine (Ad), and 0.46 and 0.93 μM/L kinetin (Kin) resulted in the best shoot multiplication and elongation. In addition, the combination of 0.46 μM/L Kin, DPU, or Ad with gibberellic acid, in particular, 0.46 μM/L Ad + 0.58 μM/L GA₃ and 0.46 μM/L Kin + 1.15 μM/L GA₃, led to better bud and shoot multiplication. Moreover, the integration of the combinations of 0.46 μM/L Kin and auxins, namely 0.46 μM/L Kin + 2.85 μM/L indole-3-acetic acid (IAA), 0.46 μM/L Kin + 2.85 or 5.71 μM/L indole-3-butyric acid (IBA), and 0.46 μM/L Kin + 0.3 or 0.57 μM/L 1-naphthaleneacetic acid (NAA), in the culture medium led to better root development and optimized aerial growth. Finally, the in vitro plants from the medium containing N₃₀K + 0.46 μM/L Kin + 2.85 μM/L IAA were successfully acclimatized; these plants served as a source for repeating in vitro culture.

• Journal of Korean Society of Forest Science
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• v.86 no.3
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• pp.391-398
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• 1997

Present study describes a method on the application of efficient tissue culture systems for the micro-propagation of juvenile and mature sawtooth oak(Quercus acutissima). Nodal segments with axillary buds were used as initial explant sources. WPM(Woody Plant Medium) was the best in growth and proliferation of shoot among the media tested. Although the single effect of zeatin revealed on two dorminant shoot elongation with normal growth until the elevation of levels up to 3.0mg/l, BAP($N^6$-benzyl amino purine) usually showed better response than zeatin on shoot multiplication and/or elongation. In addition, the incorporation of BAP and zeatin onto the culture media represents more effectiveness in shoot proliferation and its growth. Optimum concentrations of BAP and zeatin were 0.5 and 0.05~1.0mg/l, respectively. Ninety percent of the proliferated shoots was rooted on half-strength GD (Gresshoff and Doy, 1972) medium containing 0.5mg/l IBA(indole butyric acid) in 4 weeks after culture. More than 70% of the rooted plantlets survived after 5 months of transplanting into artificial soil mix containing equal amount of peatmoss and perlite. Among 27 plus tree clones which were grafted twice onto the juvenile rootstocks, only 4 clones revealed the possibility for shoot multiplication through tissue culture system. The capacity for the micropropagation using mature explant sources was highly depended on clonal differences compared with those of octet age. More than 90% of rooting ratio was obtained from the best responding clone. Among the 7 rooting media tested, GD medium was the best far rooting. The most effective rooting was obtained on half-strength GD medium containing 0.2 to 2.0mg/l IBA. More than 60% of rooted plantlets survived after 5 months of transplanting into the artificial soil mix.

• 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.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Journal of Bio-Environment Control
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• v.20 no.4
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• pp.346-351
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• 2011

This research was done to identify the mass propagation method in winter daphne (Daphne odora Thunb) using its softwood cuttings and to investigate its plant characters established at different light and soil conditions as an indoor plant. Cuttings from winter daphne were taken and grown in different treatment consisted of rooting media (perlite, vermiculite, perlite + vermiculite (1 : 1) and commercial horticulture media soil), indole butyric acid (IBA) hormone concentrations (0, 100, 500, 1000 ppm and Rootone) and date of cutting. Transplants were grown at different light intensities (100, 1000, 2500 lux and control) and growing media. Results showed that cuttings grown in perlite + vermiculite (1 : 1) gave higher percentage (100%) rooting. Cuttings treated with Rootone and IBA 100 ppm showed good rooting growth and cutting taken in June, 25 gave the highest rooting (96.7%). The best plant growth obtained at 1000 lux (56~60 ${\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) followed by 2500 lux (125~130 ${\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$) and scoria mixed with commercial horticulture media soil showed better growth of transplants.

• Journal of Plant Biotechnology
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• v.46 no.4
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• pp.310-317
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• 2019

Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica 'Fuji') leaf explants were developed in the present study. The effects of different basal media, types and concentrations of carbon sources, and concentrations of plant growth regulators were evaluated to determine the optimal shoot regeneration conditions for 'Fuji' apple leaf explants. On different treatments involving combinations of basal media, LS and N6 media, and different types and concentrations of cytokinins, 6-benzyl-adenine (BA) and thidiazuron (TDZ), shoot regeneration rates were the highest in the N6 medium combined with BA. Among the plant growth regulator and carbon source combination treatments, 5.0 mg/L BA, and 0.1 mg/L α-naphthalene acetic acid (NAA) with 40 g/L sorbitol was the optimal combination for shoot regeneration. In addition, the optimal sorbitol concentrations for shoot regeneration were 40 g/L and 60 g/L. The highest regeneration (81.8%) was achieved using 40 g/L sorbitol. The regenerated shoots elongated and rooted on rooting medium, consisting of 1/4 MS medium with 0.2 mg/L indole-3-butyric acid (IBA). The plantlets were acclimatized and the regenerated plants exhibited normal phenotypes.

• Journal of Plant Biotechnology
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• v.43 no.1
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• pp.104-109
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• 2016

Rosa rugosa is a medicinal, ornamental, and edible plant native to Eastern Asian countries, including Korea, Japan, and China. The aim of this study was to establish a system for biomass production and secondary metabolite accumulation during in vitro culture and acclimatization of Rosa rugosa. The highest rate of multiple shoot proliferation was achieved with $8.8{\mu}M$ benzyladenine (BA) (83.3%). However, the number of shoots (14.4 per explant) at $4.4{\mu}M$ BA was higher than that at $8.8{\mu}M$ BA. Compared to BA, a combination of thidiazuron (TDZ) and indole butyric acid (IBA) exhibited significantly lower shoot induction, with only 50.0~79.2% and 4.2~16.7% relative shoot formation, respectively. During acclimatization, shoots were sampled every week and their total phenolic contents were analyzed. Among various growth factors, fresh weight showed the most dramatic increase from the 3rd week (88.0 mg/plant) to 4th week (132.7 mg/plant). Total phenolics and flavonoids contents were the highest at $1^{st}$ week of acclimatization. Depending on developmental stages, total phenolics and flavonoids contents were higher in 1-yr-old shoots grown ex vitro than in those of older field-grown or in vitro-grown plants. Amongst different ages of field grown plants, 6-year-old plants, the oldest in this study, showed the lowest content in total phenolics.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.213-221
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• 2022

Wild garlic (Allium victorialis var. platyphyllum, AVVP) is a nontimber forest product used as an edible and medicinal vegetable. AVVP is usually propagated form offspring bulbs but it takes a long time to harvest. Using tissue culture technology could overcome this problem. This study investigated the optimal conditions for shoot multiplication, root growth, and plant growth by scooping AVVP bulbs. AVVP bulbs harvested from Ulleung Island, Korea, the main producer of AVVP, were surface-sterilized and used for in vitro propagation. Shoot multiplication was performed by the scooping method. More than five multiple shoots were induced from scooped tissue in Quoirin and Lepoivre (QL) medium containing plant growth regulators (PGRs); the maximum number of multiple shoots were induced from scooped tissue in QL medium containing 0.45 μM thidiazuron (TDZ) after 16 weeks of culture. Roots were induced directly at the base of the shoots in all treatments. In vitro rooting depended on the type of PGRs, and the best root-inducing treatment was QL medium containing 9.84 μM indole-3-butyric acid (IBA). Plants with in vitro roots were transferred to pots containing artificial soil and successfully acclimatized for 4 weeks. The acclimatized plants showed a survival rate of 80% after 20 weeks and gradually promoted growth depending on the acclimatization period. The results of this study will be of great help to AVVP dissemination through sustainable mass propagation.

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.42-48
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• 2017

This study was conducted to establish the optimal condition for in vitro propagation of Disporum sessile. In the experiments with different kinds of media (MS, B5 and WPM) and explant types for shoot induction and elongation, the highest number of shoot inductions (2.5/explant) was shown when the axillary bud explants were cultured on MS medium without plant growth regulators (PGRs). The best shoot elongation (7.2 cm) was also obtained when the apical buds were cultured on MS medium. The effect of BA pulse treatments with in vitro shoots was also examined. The highest in shoot induction (2.29/explant) and elongation (7.28 cm) was observed when the shoots from axillary buds were cultured on the media without PGRs. However, it decreased with increasing duration of BA pulse-treatment. The highest rooting rate (100%) and number of root inductions (21.3/explant) were achieved with 1.0 mg/L indole-3-butyric acid (IBA) treatment, whereas no differences were observed by different shoot types. The regenerated in vitro plantlets were hardened and successfully established in soil.