• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.269-273
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• 2003

In order to develop an efficient micropropagation protocal for Eucalyptus pellita, on in vitro culture system has been was established by inducing axillary buds from greenhouse stock materials. Among 6different media tested, DKW medium was the best ot induce bast induce both shoot proliferation and growth. Average number of proliferated shoots of 403per explant was obtained at the concentration of 0.1mg/LBA. Most of the stem materials excreted phenolic compounds at the proximal part of the explant and caused darking of the media. Therefore, it was necessary to transfer frequently to a fresh medium and/or to add activated charcoal at the concentration of 0.02％(w/v). Generally on vitro roots were formed easily on 1/2DKW medium with NAA treatment. All the explants rooted at the medium containing 0.2mg/L NAA and displayed vigorous root growth in vitro culture conditions. After transferred to an artificial soil mixture (peatmoss: vermiculrite: perlite, 1:1:1, v/v/v) in the greenhouse, most rooted plantlets survived well without any morphological abnormalities. The results show that the species can be micropropagated effectively by the application of axillary bud culture system.

• Journal of Plant Biotechnology
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• v.5 no.2
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• pp.115-119
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• 2003

To determine the appropriate concentrations of nutrients and growth regulators for shoot proliferation and root initiation, several rose hybrid tea cultivars were cultured. Cultured shoot tips and lateral buds from different cultivars proliferated multiple shoots on Murashige and Skoog (MS) medium supplemented with 0 to 4 mg/L BA and 0 to 0.05 mg/L NAA. The ability of the explants to proliferate shoots and initiate roots was affected by genotype, the nodal position of explant, the strength of MS basal medium and growth regulators used. The buds nearest the apex exhibited the slowest rate of development. Most cultivars had the highest shoot proliferation when cultured on MS medium with 2 mg/L BA and 0.01 mg/L NAA, but the degree varied by cultivars. Root development was enhanced by lowering the concentration of MS salts.

• Journal of Korean Society of Forest Science
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• v.96 no.2
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• pp.178-182
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• 2007

Rauvolfia serpentina (L.) Benth. ex Kurz is a medicinal plant and an endangered tropical rainforest plant species. Since the field cultivation that aims to fulfill the industrial needs is never accomplished, tissue culture appears to be the most feasible way to improve the quality and quantity of R. serpentina. This experiment used two kinds of explants (roots and shoots) to induce optimal root formation in different combinations of auxin and photoperiod. Each explants exhibited different responses on given treatments. Differentiated root could be produced from explants cultured in IBA 20 mg/L with and without light. The highest number of roots, root length and root weight induced from shoot explants were effective on MS medium containing IBA 20 mg/L and incubated under dark condition, while highest total weight (callus and root) from root explants cultured on MS medium supplemented 10 mg/L IBA and 10 mg/L NAA and incubated under day length (11/13 hr). The root induced from shoot explants produced the highest major alkaloid content. The highest content of ajmaline (2.17 ppm fresh weight) and reserpine (1.30 ppm fresh weight) were observed in shoot explants cultured in MS medium containing combination of IBA 10 mg/L and NAA 10 mg/L and incubated under dark condition, yohimbine (1.47 ppm fresh weight) was in the shoot explants cultured in MS medium containing NAA 20 mg/L and incubated under day length, while serpentine was absent.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.1-6
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• 2000

This study was carried out to find the effects of plant growth regulators on callus formation, rooting and shooting from cotyledon explant in oriental me]on. Various combinations of 0.1 mg/L auxins (IAA, NAA) and 0.5, 1.0. 1.5, 2.0 mg/L cytokinins (BA, kinetin, zeatin) were treated to the MS basal medium, respectively. Callus was induced mort effectively as 2,437.0 mg (FW)/explant in MS medium supplemented with 0.1 mg/L NAA and 2.0 mg/L BA, but that was non-embryogenic callus as colored yellow white and broke easily. Root was induced most effectively at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L NAA and 0-5 mg/L kinetin. Shoots formed on cut part of vein at a frequency of 98.0% in MS medium supplemented with 0.1 mg/L IAA and 2.0 mg/L BA, that were multiple shoots. in case of its concentration, BA and lower concentration of IAA and NAA (0.01 and 0.05 mg/L). respectively. shooting ratio was not increased. The result of treatment with BA 0-5 mg/L and IAA 0.1 mg/L, callus induced at a week, and shoot start to form multiple shoots about 3 weeks after inoculation. After 2 times subculture as 2 weeks intervals, divided shoots rooted and developed into intact plantlets at 10 weeks and then that grown normally on pots after acclimatization.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.155-161
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• 2008

This paper reported the establishment of mass production system of adventitious roots of Codonopsis lanceolata through shake flask and bioreactor culture. Induction of adventitious roots was started from the explants of leaf, stem and root on 1/2 strength Murashing and Skoog (MS) solid medium. Stem segments were the best explants for induction of adventitious roots compared to leaf and root segments. Among the different auxins tested (NAA, IBA and IAA), number of adventitious root per explant was highest on solid medium with 1.0 mg/L IBA, and produced $9.9{\pm}1.2$ roots per explant. However, growth of adventitious roots was fast in the presence of IBA at low concentration (0.1 mg/L). In shake flask culture, maximum production of adventitious roots (fresh weight) was obtained in half-strength MS medium compared to full-strength and one-third MS medium. When the adventitious roots produced in shake-flask culture were transferred to 5 L air-lift bioreactor, 16 times of fresh weight increase was gained after one month of culture. These results indicate that this protocol for the production of C. lanceolata adventitious roots can be applied to large scale culture for practical application.

• Plant Biotechnology Reports
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• v.4 no.4
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• pp.303-309
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• 2010

Endoreduplication is a developmental process that is unique to plants and occurs in all plants. The present study aimed to assess endoreduplication in various explant tissues and regenerated somatic embryos of Doritaenopsis. We further investigated the effects of light quality on endoreduplication and somatic embryo proliferation. To this end, we studied endoreduplication in leaves and root tips from regenerated plantlets and somatic embryos and in developing somatic embryos under 4 types of lighting conditions: red light, red + far-red light, red + blue light, and white light. We found that the degree of endoreduplication varied in different explants, and that the choice of explants used also influenced the ploidy levels of the newly regenerated somatic embryos. The DNA content of the leaf (2C-8C) was less than that of the root tip (2C-16C) and somatic embryo (2C-64C). In terms of light quality, the combination of red and far-red light produced the highest number of somatic embryos, while maintaining a low degree of endoreduplication. The data obtained indicate that this light combination stimulates somatic embryogenesis in Doritaenopsis and may exert some control on endoreduplication during cell division. These findings can be applied to achieve a reduction in somaclonal variations for the purpose of mass proliferation and genetic improvement.

• Journal of Plant Biotechnology
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• v.50
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• pp.108-114
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• 2023

In vitro conservation is one of the most effective strategies for rare plant protection, especially for orchid species. To maximize the success rates of in vitro explant establishment (stage I) in conservation programs, the application of tissue culture additives such as Plant Preservative Mixture^TM (PPM^TM) should be emphasized. In this study, we used Dendrobium thyrsiflorum Rchb.f. (1875) seeds and seedlings as a model for the evaluation of PPM^TM's phytotoxicity in the meristematic tissues of epiphytic orchids. PPM^TM had no observable inhibitory effect on protocorm, shoot, or root development when it was supplemented at 0.1%. PPM^TM supplementation caused adverse effects on D. thyrsiflorum explants at concentrations > 0.2%. At high concentrations, young in vitro seedlings showed damage, especially at the root tissue level. Based on this model, supplementation of 0.1-0.2% PPM^TM to culture media was successfully implemented to establish in vitro cultures of other rare orchid species in our conservation program.

• Journal of Plant Biotechnology
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• v.47 no.3
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• pp.235-241
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• 2020

This is the first study to establish a complete protocol for micropropagation of Rehmannia glutinosa from root segments. The study involved investigating the effect of plant growth regulators on in vitro shoot regeneration and rooting and identifying substrates supporting survival and growth performance of ex vitro seedlings. A Murashige and Skoog (MS) medium containing 30 g/L sucrose for shoot induction and 0.2 mg/L indole-3-acetic acid (IAA), 1 mg/L 6-benzylaminopurine (BAP), and 1 g/L polyvinylpyrrolidone (PVP) for shoot multiplication resulted in the highest number of shoots per explant and shoot height. Applying a medium containing 0.5 mg/L IAA and 1 g/L PVP yielded optimal rooting of the shoots grown in vitro. Compost enriched with microbial inoculants and perlite enhanced seedling growth better than that with organic biofertilizer-free substrates (soil and sand). We recommend the continuous production of micropropagated R. glutinosa seedlings from root segments under the aforementioned conditions as a possible propagation technique for crops of this species.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.182-188
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• 2012

Iris dichotoma Pall. is an important endangered plant belonging to the family Iridaceae. A method was developed for the rapid micropropagation of I. dichotoma through plant regeneration from leaf, rhizome, and root explant-derived calli. Leaf, rhizome, and root segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; $0-3.0mg{\cdot}L^{-1}$) for callus induction. Callus production was highest at $1.0mg{\cdot}L^{-1}$ 2,4-D, where 73.8% and 45.5% of cultured rhizome and root cuttings, respectively, produced calli. The viable calli were maintained at an induced concentration of 2,4-D ($3.0mg{\cdot}L^{-1}$). They were then transferred to MS medium supplemented with various concentrations of 2,4-D ($0-3.0mg{\cdot}L^{-1}$) in combination with 6-benzyladenine (BA: 0, 1.0 and $3.0mg{\cdot}L^{-1}$) for adventitious shoot regeneration. The addition of a low concentration of 2,4-D into BA-containing medium significantly increased the frequency of shoot regeneration in leaf, rhizome, and root-derived calli. The highest number of adventitious shoots (26.4 per callus) formed at $0.5mg{\cdot}L^{-1}$ 2,4-D and 1.0 mg/l BA. For rooting of the shoots, half- strength MS medium supplemented with different concentrations of indole 3-butyric acid (IBA) $0-3.0mg{\cdot}L^{-1}$ was tested. The optimal results were observed using half-strength MS medium supplemented with $1.0mg{\cdot}L^{-1}$ IBA, on which 98% of the regenerated shoots developed roots with an average of 3.5 roots per shoot within 45 days. The plantlets raised in vitro were acclimatized and transferred to soil with 95% success. This in vitro propagation protocol will be useful for conservation and mass propagation of this endangered plant.