• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.164-170
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• 2017

A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

• KSBB Journal
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• v.7 no.2
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• pp.85-91
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• 1992

The role of NAA(1-Naphthaleneacetic acid) and BA(6-Benzyladenine) in the adventitious shoot regeneratlon from Populus leaf segments and changes in the pattern of RNA and protein synthesis were investigated. The adventitious shoot regeneration octured at the basal cut end of Populus leaf segments. This process was effected by many factors, including wounding culture conditions, light and plant growth regulators etc. The highest adventitious shoot regeneration frequency was obtained at $0.01mg/\ell$ NAA with $0.2mg/\ell$ BA. In this condition adventitious shoot starved to regenerate on the 13th day of oullure. The most optimal hormone composition for RNA and protein synthesis was $0.01mg/\ell$ NAA with $0.2mg/\ell$ BA. The content of RNA and protein was greater at the proximal part. In the course of adventitious shoot regeneration, the proteins associated with polar-regeneration appeared at the proximal part of populus leaf segment.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.157-163
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• 2010

A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.

• Korean Journal of Medicinal Crop Science
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• v.25 no.2
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• pp.95-101
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• 2017

Background: Callus cultivation has the advantage of producing a large amount of tissue of a plant in a laboratory regardless of the environment, for extracting an active substance. In the present study, callus formation was induced in the leaves of the succulent plant Adenium obesum (Forssk.) Roem & Schult. After callus cultivation, anti-inflammatory activity tests were conducted, because leaves and stems of A. obesum have been reported to possess biological activity. Methods and Results: In order to induce callus formation, various concentrations of plant growth factors, such as kinetin, naphtha-leneacetic acid (NAA), 6-benzyladenine (BA), and indole-3-acetic acid (IAA) were added to MS solid medium. The maximum callus proliferation was induced by mixed medium consisting of NAA ($2mg/{\ell}$) and BA ($1mg/{\ell}$). In addition, an elicitor was added to the medium under optimal conditions for initiating suspension culture. After suspension culturing, the activities of the callus extracts were compared and analyzed. The cytotoxicity and anti-inflammatory activity tests revealed that the anti-inflammatory activity of the callus extract and the content of phenolic compounds were elevated after treatment of the callus culture with the elicitior. Conclusions: A. obesum callus might be considered as potential source of biologically active anti-inflammatory material.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.221-224
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• 2003

Leaf explants of Ixeris sonchifolia produced adventitious shoots at a frequency of 100% when cultured on MS medium supplemented with combinations of various concentrations of 6-benzyladenine (BA) (0.44, 4.44, or 8.87 ${\mu}M$) and 0.54 ${\mu}M$ NAA, or MS medium supplemented with 22.19 ${\mu}M$ BA and 2.69 ${\mu}M\;\alpha$-naphthaleneacetic acid (NAA) after four weeks of culture. Each explants (approximately $3{\times}6mm$) produced greater than 70 shoots at a combination of 0.44 ${\mu}M$ BA and 0.54 ${\mu}M$ NAA. Leaf explants produced shoots at a frequency of greater than 80% even at as low as 0.13 ${\mu}M$ BA as the sole growth regulator. Upon transfer to one-third strength MS with 0.54 ${\mu}M$ NAA, excised adventitious shoots were rooted at a frequency of 100%. Regenerated plantlets were transplanted to potting soil and grown to maturity in a greenhouse. The competence of I. sonchifolia for plant regeneration via organogenesis appears to be greater than the competence of tobacco, currently the best model plant for organogenesis.

• Journal of Plant Biotechnology
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• v.36 no.2
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• pp.137-143
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• 2009

An efficient system for high frequency plant regeneration was established through investigating various factors such as plant growth regulator combinations, explant types and ages, and addition of $AgNO_3$ influenced on shoot regeneration in Brassica juncea L. cv. BARI sarisha-10. Murashige and Skoog (MS) medium supplemented with 0.1 mg/L NAA (1-naphthaleneacetic acid) and 1 mg/L BA (6-benzyladenine) showed the maximum shoot regeneration frequency (56.67%) among the different combinations of NAA and BA. Explant type, explant age, and addition of $AgNO_3$ also significantly affected shoot regeneration. Of the four type of explants (cotyledon, hypocotyl, root, and leaf explants)- cotyledon explants produced the highest shoot regeneration frequency and hypocotyls explants produced the highest number of shoots per explant, whereas root explants did not produce any shoot. The cotyledonary explants from Four-day-old seedlings showed the maximum shoot regeneration frequency and number of shoots per explant. Shoot regeneration frequency increased significantly by adding $AgNO_3$ to the medium. Two mg/L $AgNO_3$ appeared to be the best for shoot regeneration with the highest shoot regeneration frequency (86.67%) and number of shoots per explant (7.5 shoots). Considerable variation in shoot regeneration from cotyledonay explants was observed within the B. juncea L. genotypes. The shoot regeneration frequency ranged from 47.78% for cv. Shambol to 91.11% for cv. Rai-5. In terms of the number of shoots produced per explant, B. juncea L. cv. Daulot showed the maximum efficiency. MS medium supplemented with 0.1 mg/L NAA showed the highest frequency of rooting. The regenerated plantlets were transferred to pot soil and grown to maturity in the greenhouse. All plants were fertile and morphologically identical with the source plants.

• Horticultural Science & Technology
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• v.35 no.2
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• pp.252-264
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• 2017

Tea is one of the most consumed beverages worldwide and the relatively high levels polyphenols is benefit for health. In this study, we developed an efficient system for proliferation of callus from 'Anji Baicha', a cultivar of tea (Camellia sinensis). Callus tissue was initially induced by culturing leaf explants on medium containing different plant growth regulators. For callus induction, thidiazuron (TDZ) was more effective than 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}-naphthalene$ acetic acid (NAA), and $N^6-benzyladenine$ (BA). The frequency of callus induction from leaf explants reached 90.21% on $1.0mg{\cdot}L^{-1}$ TDZ and the developed callus was reddish and friable. We also tested the effect of different concentrations of NAA, 2,4-D, indole 3-acetic acid (IAA), BA, and TDZ, alone and in combinations, on callus proliferation. Medium supplemented with TDZ in combination with IAA was suitable for callus proliferation and accumulation of tea polyphenols. The growth index value and tea polyphenol content of callus cultured on MS medium containing $0.5mg{\cdot}L^{-1}$ TDZ and $1.0mg{\cdot}L^{-1}$ IAA was maximally 1,351% and 23.24%, respectively, and the relative abundance of epicatechin was as high as 17.449%. We also measured the antioxidant activity of all samples and the callus with the highest tea polyphenol content also exhibited high potential radical scavenging activity.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.1-6
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• 1994

Cotyledonary and hypocotyl explants of melon seedlings were cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and benzyladenine (B.A).Up to 22% of cotyledonary explants and 7%, of hypocotyl explants, respectively: Produced somatic embryos through intervening two types of calli: bright yellow compact (BYC) callus and pale-yellow compact (PYC) callus. BYC callus was capable of producing somatic embryos at initial culture, but it became necrotic as subrulhues proceeded. In contrast UC callus was incapable of producing somatic embryos during initial culture (first 6 weeks), but it became bright-yellow friable (BYF) callus with forming a few globular embryos after 2 months of subculture, indicating that the callus turned embryogenic. The embryogenic capacity of BYF maintained for over one year when the callus was sucultured at 4-week interval. Upon transfer onto MS basal medium the callus gave rise to numerous somatic embryos and subsequently converted to plantlets. Plantlets were transplanted to potting soil and grown to maturity in the phyotron.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.161-166
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• 2007

In vitro shoot regeneration and multiple shoot induction has been obtained from the stolen node explants in Eremochloa ophiuroides (Munro) Hack. The highest number of shoots ($10.66{\pm}0.21$) was observed from initial explants after one month culture duration on Murashige and Skoog (MS) medium containing 6-benzyladenine (BA: 0.5 mg/l). First generation shoot was excised and sub-cultured on the same fresh media for further multiplication of shoots. An enhanced number of second round shoots ($15.33{\pm}0.21$) was obtained compared to the initial culture media containing BA (0.5 mg/l). The number of shoots/stolon node was higher among all the concentrations of BA than kinetin (KN). In vitro regenerated shoots were successfully rooted in the phytohormone free MS medium. Plantlets generated with roots were transferred to pots containing compound mixture of soil and kept in green house conditions. Acclimatized plants showed 100% survival rate with normal morphology in green house conditions. The present study demonstrates the effect of explant and different plant growth regulators towards in vitro response in E. ophiuroides. Moreover, the study reveals the effect of cytokinin on induction of shoot number per stolen node explant in E. ophiuroides.

• Journal of Plant Biotechnology
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• v.46 no.4
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• pp.291-296
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• 2019

This study was conducted to determine the effect of a light-emitting diode (LED) on in vitro shoot growth and rooting in teak (Tectona grandis L.). In the experiments with apical bud explants, the greatest shoot elongation (3.2 cm) occurred when they were cultured on DKW medium under 50% blue and 50% red LED mixture (BR), whereas no differences in growth were observed in different light sources (florescent light [F] or BR) or media (MS or DKW). The highest number of shoot multiplication (2.4/explant) or elongation (4.94 cm) was achieved with 0.5 or 1.0 mg/L 6-Benzyladenine (BA) treatment under BR. In addition, the best rooting rate (93.8%) or root length (1.3 cm) was recorded with 0.5 mg/L indole-3-butyric acid (IBA) treatment under BR, and the highest root induction (3.1/explant) was observed in 0.2 mg/L IBA under BR. The in vitro rooted plantlets were hardened and survived well on soil.