• Journal of Plant Biotechnology
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• v.50
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• pp.169-175
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• 2023

A plant regeneration system was developed through shoot organogenesis from in vitro leaf explants of Chrysanthemum morifolium 'Ohblang'. The effects of different concentrations of plant growth regulators and AgNO₃ on efficient shoot regeneration and inhibition of browning were evaluated in chrysanthemum. The explants were cultured on MS shoot induction medium supplemented with 12 combination treatments of 6-benzyladenine (BA) 0.5, 1.0 and 2.0 mg/L, and α-naphthaleneacetic acid (NAA) 0.2, 0.5, 1.0 and 2.0 mg/L in darkness for 6 weeks and cultured under a 16/8 h photoperiod for 6 weeks. The highest shoot regeneration was obtained from the explants cultured on the medium with 1.0 mg/L BA and 1.0 mg/L NAA. Based on this result, AgNO₃ was added to a shoot induction medium containing MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA, 30 g/L sucrose, and 6 g/L agar to reduce browning of chrysanthemum leaf explants. In the control treatment without AgNO₃, leaf explants turned brown at the cut edge; however, browning was not observed in AgNO₃ treatments. Shoot organogenesis was higher at low concentrations of AgNO₃ and decreased with an increase in AgNO₃ concentration. The explants cultured on shoot induction medium (MS salts, vitamins, 1.0 mg/L BA, 1.0 mg/L NAA) with 1 mg/L of AgNO₃ produced the highest shoot regeneration with 2.6 shoots per explants and a browning index of 0.7. When the regenerated shoots were detached from the explants and cultured on MS medium, the shoots were elongated and rooted successfully.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.4
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• pp.331-335
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• 2013

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) $2.0mg{\cdot}L^{-1}$, benzyladenine (BA) $1.0mg{\cdot}L^{-1}$ and 3% sucrose. Friable callus was transferred to solidified MS media containing BA ($1.0mg{\cdot}L^{-1}$) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid ($2.0mg{\cdot}L^{-1}$), Kinetin ($1.0mg{\cdot}L^{-1}$), BA ($1.0mg{\cdot}L^{-1}$). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

• 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.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.

• Korean Journal of Environmental Agriculture
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• v.16 no.3
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• pp.227-232
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• 1997

Visible injury appeared 7 days after ultraviolet-B(UV-B) irradiation, but did not show any significant decline of growth in cucumber plant. However the growth of the first leaves of fertilized plants was suppressed by UV-B irradiation. Especially the most effective growth retardiation appeared when supplied with nitrogen rather than phosphate and potassium. These results suggest that UV-B may play an important role in inhibiting nitrogen metabolism. Therefore we examined the effect of activity of nitrate reductase, and found that the nitrate reductase activity of the first leaves was increased by UV-B irradiation for 7 days and fertilization. We examined the effect of plant hormone on the inhibition of growth in the first leaves. Benzyladenine promoted the growth of discs excised from the first leaves by fertilization and without UV-B, but did not promote the growth of leaf discs from UV-B irradiated plants. We conclude that the UV-B-induced decrease in the growth of the first leaves could be related to reduction in sensitivity to plant hormones.

• FLOWER RESEARCH JOURNAL
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• v.18 no.2
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• pp.87-92
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• 2010

This research was aimed to extend the vase life, improve the cut flower quality and promote flowering rate of bulbous cut Iris 'Blue Magic'. Three different concentrations of 1-MCP ranging from 250 to $750nL{\cdot}L^{-1}$ were treated on cut Iris flowers for the vase life elongation. Several effects of 1-MCP treatment has shown such as early flowering as one day but vase life showed no significant differences comparing to the untreated control. Flowering rate was 75% or more in the treated ones as compare to 64.2% in untreated control. Especially, in a treatment with $250nL{\cdot}L^{-1}$ 1-MCP for 12 hours it showed all flowering. In many post-harvest experiments of cut flowers, ethylene production from flower organ has considered as most critical factor on vase life. Ethylene production from flower organ was measured by gas chromatography (GC) and it showed that there is no direct relation between flower longevity and emission of the ethylene gas in cut Iris. In comparison with the untreated control, ethylene generation rather seems to be increased as 1-MCP treatment increased. There was a synergy effect when 1-MCP and BA applied simultaneously in which flowering and vase life were fastened and also extended, respectively. Therefore, treatment of 1-MCP for cut Iris at flower early harvest is able to improve the flower quality both by diminishing non-flowering rate and by extended flower longevity.

• 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.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.