• Journal of Plant Biotechnology
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• v.42 no.3
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• pp.257-264
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• 2015

To determine whether metabolite fingerprinting for whole cell extracts based on Fourier transform infrared (FT-IR) spectroscopy can be used to discriminate and compare metabolic equivalence, standard medicinal parts from four medicinal plants (Cynanchum wilfordii Hemsley, Atractylodes japonica Koidz, Polygonum multiflorum Thunberg and Astragalus membranaceus Bunge) and their in vitro-produced adventitious roots were analyzed by FT-IR spectroscopy. The principal component analysis (PCA) and partial least square discriminant analysis (PLS-DA) from the FT-IR spectral data showed that the whole metabolic pattern from Cynanchum wilfordii was highly similar to Astragalus membranaceus. However, Atractylodes japonica and Polygonum multiflorum showed significantly different metabolic patterns. Furthermore, adventitious roots from Cynanchum wilfordii and Astragalus membranaceus also showed similar metabolic patterns compared to their standard medicinal parts. These results clearly show that mass proliferation of adventitious roots may be applied to aquire novel supply of standard medicinal parts from medicinal plants. However, the whole metabolic pattern from adventitious roots of Atractylodes japonica and Polygonum multiflorum were not similar to their standard medicinal parts. Furthermore, FT-IR spectroscopy combined with multivariate analyses established in this study may be applied as an alternative tool to discriminate the whole metabolic equivalence from several standard medicinal parts. Thus, we suggest that these metabolic discrimination systems may be applied for metabolic standardization of herbal medicinal resources.

• Korean Journal of Medicinal Crop Science
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• v.8 no.1
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• pp.14-20
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• 2000

This experiment was conducted to investigate the effects of length of storage period under low temperature, $CO_2$ enrichment and addition of plant growth regulators in Murashige and Skoog medium on the plant regeneration of Korean ginseng (Panax ginseng C. A. Meyer). Seeds were treated for 60 and 80 days respectively under $5^{\circ}C$ environment. 2500ppm of $CO_2$ was enriched by ventilation. Three plant growth regulators added to the medium were Indolbutyric acid, Benzyladenin and Gibberellic acid (GA3). The result indicated that : The capacity of differentiation was higher in the aged cotyledons from the seeds treated for 80 days under low temperature condition than in those treated for 60 days. $CO_2$ enrichment had stimulating effects on the growth and development of shoot primordium significantly but less effects on the formation of adventitious buds. From one zygotic embryo hundreds of plantlets were differentiated. $CO_2$ enrichment had effects on the formation of both indirect somatic embryo and direct somatic embryo. Indirect somatic embryo showed little growth and differentiation, being undifferentiated vascular stele and epicotyl. Direct somatic embryos were formed on the epidermis of backside basal part of cotyledon. Those embryos developed to whole plant having latent bud.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.261-266
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• 1995

This study was carried out to develop new varieties which are dwarf and tolerant to winter cold in the Aurantioideae by intergeneric crossing. to do that, the reciprocal crosses of Hwanggeumyooza and trifoliate orange, yooza and trifoliate orange were done and in vitro immature ovule culture of their hybrid was carried out .The callus formation from immature ovule was good in order of Hwanggeumyooza, Hwanggeumyooza $\times$ tifoliate orange, yooza, and trifoliate orange and best at 1 to 3 mg/L NAA+0.5mg/L zeatin on MT medium. In vitro germination percentage of 20week old hybrid of Hwanggeumyooza $\times$ tifoliate orange and trifoliate orange $\times$ Hwanggeumyooza were 41.3% and 37.7, respectively. The phenotype of hybrid (95%) of Hwanggeumyooza $\times$ trifoliate orange and that (100%) of trifoliate orange $\times$ Hwanggeumyooza were similar to that of trifoliate orange. After Hwanggeumyooza was pollinated by pollens of trifoliate orange, the pollen tubes grew on stigma after 3h of pollination and entered into micropyle after about 24~28 h. One gamete in pollen was fused with polar nuclei after 2 days and other one fused with egg nucleus at 3days after pollination. The fruit set percentage by intergeneric crossing was 14.0% in Hwanggeumyooza $\times$ trtfoliate orange and 17.5% in trifoliate orange $\times$ Hwanggeumyooza. The fruit set percentages of Hwanggeumyooza. and trifoliate orange were 34.2% and 39.5% by artificial self-fertilization, 34.2% and 39.5% by artificial cross fertilization, 3.1% and 1.4% by parthenocarpy and 13.0% and 3.0% by natural fertilization, respectively. The somatic and gametic chromosome numbers of Hwanggeumyooza, yooza, and trifoliate orange were 2n=18 and n=9.

• Research in Plant Disease
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• v.19 no.4
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• pp.288-293
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• 2013

In worldwide, viral diseases of apple plants has caused the serious problems like reduced production and malformation of fruits. Also, the damages of apple plants by virus and/or viroid infection (Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple mosaic virus, and Apple scar skin viroid) were reported in Korea. However there is few report about the protection approach against the infection by apple viruses. Therefore, this paper introduced the experimental protocol for the development of virus-free apple cultivars (Danhong, Hongan, Saenara, Summerdream). Apple plants were treated at $37^{\circ}C$ for 4 weeks and shoot tips were cultured in vitro. After heat treatment, the detection of apple viruses was performed by RT-PCR using virusspecific detection primers in new apple cultivars. With the heat treatments followed by in vitro shoot tip culture, the proportion of virus-free stocks of 'Danhong', 'Hongan', 'Saenara', and 'Summerdream' was 28%, 16%, 12%, and 12%, respectively. Taken together, this approach can be a good tool for production of virus-free apple stocks.

• Journal of Bio-Environment Control
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• v.24 no.3
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• pp.167-172
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• 2015

The aim of this study was to develop an effective production system of blueberry plants by using tissue culture technique. Murashige and skoog medium (MS) and woody plant medium (WPM) were compared for shoot formation of highbush blueberries. Also medium supplemented with zeatin/2-isopentenyl adenine (2iP)/benzyl aminopurine (BA) (1, 2/10, 15/4, $6mg{\cdot}L^{-1}$)and zeatin/2iP/BA (0.5/10, 15/$0.05mg{\cdot}L^{-1}$) as plant growth regulators to determine the effect of shoot formation and shoot proliferation, respectively. The shoot explants cultured on WPM showed higher shoot formation rates, more number of nodes, and longer root length than those on MS medium during the primary culture. Shoots were not formed when the explants were cultured on the medium without plant growth regulators or on only BA. The shoot explants cultured on the medium supplemented with 2iP showed low rates of shoot formation. On the other hand, zeatin was the most effective for shoot formation and growth of the explants. Also influence of different cytokinins (zeatin, 2iP) on the shoot proliferation of subcultured shoot explants was studied. There was no significant difference among the different concentrations of zeatin in the rate of shoot formation and number of shoots. However at higher concentration of zeatin, number of nodes was increased, and shoot length was shorted. The proper concentrations of zeatin for shoot propagation in subculture were found to be $0.5mg{\cdot}L^{-1}$ and $1mg{\cdot}L^{-1}$.

• Korean Journal of Plant Resources
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• v.16 no.2
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• pp.160-167
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• 2003

In order to investigate the effects of developmental stage of embryos and plant growth regulators on mass production of Zantedeschia spp. Southern Light, immature zygotic embryos of Zantedeschia spp. Southern Light were cultured on Murashige and Skoog(1962) basal media or containing 2,4-D, NAA and BA. Globular embryos did not grow on any of the 2,4-D, NAA and BA combinations. The most suitable stage of immature zygotic embryo culture on the induction callus and multiple shoot was at early cotyledonary embryo stage, and at this stage of embryos were germinated up to 87.5%. The whitish watery callus and yellowish compact nodular callus produced on all 2,4-D, NAA and BA media. The best combination for inducing embryogenic callus was 0.5 mgL NAA and 1.0 mg/L BA. Whitish watery calli have been subcultured for more than 8 months and have retained their producing ability, Plant regeneration was only obtained by direct shoot development and yellowish compact nodular calli. Abundant plantlets were regenerated from cotyledonary stage of embryo culture on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA. Supplementation of the media with 10％ coconut water showed as the best concentration for plant differentiation from direct developed of shoots. The number of regenerated plants from one embryo could be seperated 25-35s plantlets. All yellowish compact callus-derived plantlets were transferred to pots containing a mixture of vermiculite, perlite and sand(1:1;1 v/v) and 100% of divided plantlets were phenotypically normal.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.336-339
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• 2003

Hydrocotyle maritima Honda used as medicinal plants for a hemostatic agent was investigated for in vitro regeneration. The petiole explants of H. maritima were cultured on callus induction medium containing growth regulators ($0{\sim}5\;mg/l$, NAA and $0{\sim}5\;mg/l$ 2,4-D) either in single or in combination with $0.1{\sim}2\;mg/l$ BA for 6 weeks. Although single treatments of 2,4-D or NAA resulted in callus formation, the best results were combination of $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA. $5\;mg/l$, NAA and $0.5\;mg/l$, BA, respectively. The highest number of shoot (12 shoots per callus) was achieved with $3\;mg/l$, Kinetin. Also, when pieces of embryogenic callus induced on the medium supplemented with $0.5\;mg/l$, 2,4-D and $0.5\;mg/l$, BA were subcultured on hormone-free medium, somatic embryos were differentiated and developed further into welldeveloped plants.

• Korean Journal of Plant Resources
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• v.27 no.5
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• pp.540-545
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• 2014

The Polygonatum odoratum cv. Gungangbeaksea, bred in Gyeongsangnam-Do Agricutural Research & Extension Service, was cultured in vitro for micropropagate rapidly through the culture of rhizome explants ($5{\times}5mm$). The $7{\times}7mm$ explants of adventitious multi-bud clusters (AMC), obtained through the culture of rhizome explants (MS + 3.0 mg/L BA) were cultured on MS media with BA and TDZ. The shoot multiplication was favorable on the MS medium containing 3.0 mg/L TDZ with 2.8 in shoot number. But the formation of AMC was low in all media tested. The explants of AMC were cultured on MS media containing 1.0~5.0 mg/L TDZ and NAA to multiplicate AMC more. The formation of AMC was a little more stimulated on combined MS media of TDZ and NAA, than that with TDZ alone. The multiplication of shoots and AMC was favorable on MS media with 3.0 mg/L TDZ and 5.0 mg/L NAA, and 5.0 mg/L TDZ and 3.0 mg/L NAA. As the concentration of MS salts increased, the formation of AMC was decreased. But the formation of AMC was more stimulated, as the concentration of sucrose increased to 7%. Therefore, the multiplication of shoots and AMC was suitable on media containing 3.0~5.0 mg/L TDZ and NAA, and 7% of sucrose. The explants of AMC were rooted on media with 3.0 mg/L IBA, or 2.0 mg/L NAA with more than 80% in rooting ratio. The plantlets were treated at $5^{\circ}C$ for 8 weeks, and cultured ex vitro for 8 weeks. The survival ratio of plantlets were 100% in vermiculite, and the mixed soil with perlite 1 volumn and vermiculite 1 volumn.

• Journal of Plant Biotechnology
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• v.47 no.3
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• pp.242-247
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• 2020

This experiment was performed to determine the concentration of coconut water that is most effective in promoting the growth of 'Maehyang' strawberry in tissue culture. Cultivars in this experiment consisted of 'Maehyang' grown in the presence of 0, 20, 40, 60, 80, 100 ml·L^-1 of coconut water which was added to a medium mixed with BA 0.5 mg·L^-1 and IBA 0.1 mg·L^-1. Morphological variation tests and SSR detection with coconut water were conducted to determine variations in proliferation. The proliferation rate of 'Maehyang' strawberry improved with the coconut water treatment compared to non-treatment. In particular, the proliferation rate with 100 ml·L^-1 coconut water treatment increased by about 4 times. When 'Maeyang' was treated with 100 ml·L^-1 of coconut water, two morphological variants occurred without genetic variation. Therefore, the results suggest that 100 ml·L^-1 of coconut water can be used to mass-produce "Maeyang" strawberry without causing genetic variations.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.281-285
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• 2002

Whole plant cultures of Centella asiatica (L.) Urban in vitro were established and the effects of basal media, some macro elements and sucrose concentration on productivity of triterpene glycosides (madecassoside and asiaticoside: M$\varepsilon$A) were investigated. Among the media (MS, B5, RCM) tested, MS and 0.5 RCM medium were the best for plant growth and M$\varepsilon$A production, respectively. However, taking into account the M$\varepsilon$A productivity, B5 medium was superior (M$\varepsilon$A: 14.28 mg/g dry wt.). Major macronutrients of B5 medium adjusted with the concentration of 25 mM KNO$_3$,1 mM NaH$_2$PO$_4$, 1 mM CaCl$_2$ and 1~10 mM MgSO$_4$, caused elevated or optimized levels of M$\varepsilon$A. On sucrose concentration, the highest yields of M$\varepsilon$A were obtained from 6% sucrose.