• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.230-239
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• 2022

Holostemma annulare (Family Asclepiadaceae) is an invaluable vulnerable medicinal plant; the root tubers are used in Ayurveda medicine and by folk healers to treat various ailments. In this study, Schenk and Hildebrandt medium fortified with the cytokinins 6-benzyl adenine, kinetin, and auxins, including indole 3-butyric acid, indole 3-acetic acid, α-naphthaleneacetic acid, and 2,4-dichlorophenoxyacetic acid, were checked for their efficiency on root tuber induction from different explants. Adventitious root tubers were more successfully induced from in vitro leaf segments and shoots when cultured in Schenk and Hildebrandt medium supplemented with 0.5 mg/l of α-naphthaleneacetic acid. In addition, preliminary phytochemical analysis of in vitro root tubers and identification of different secondary metabolites were conducted. Thin layer chromatography and high performance thin layer chromatography analysis of the crude methanolic extracts of the in vitro root tuber identified the presence of lupeol, a bioactive triterpene. Adventitious root tuber induction offers a novel method for the in vitro production of bioactive metabolites that can be scaled up by bioreactors, thus ensuring the conservation and sustainable utilization of H. annulare. The study warrants further scale-up production and pharmacological investigation that can be extended for pharmaceutical needs.

• Korean Journal of Medicinal Crop Science
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• v.14 no.4
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• pp.225-228
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• 2006

To acquire the normal regeneration of plantlets, we investigated combinations and concentrations of plant growth regulations for optimal conditions of adventitious root formation. Based on the previous study, we performed callus and shoot induction. When induced shoot was transferred into a rooting medium containing plant hormones, it wilted and died. Thus, the shoot proliferated on 1/2 MS medium for 10 days and was then treated with MS medium supplemented with 3.0 mg/L NAA for 3 days. Adventitious root formations were observed after shoot planlets were transferred to 1/3 MS medium. The concentrations of salt and sucrose were gradually reduced in MS medium and the rooted plantlets were transferred for acclimatization into a mixture of peatmoos : perlite (3 : 2).

• Journal of Life Science
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• v.25 no.4
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• pp.390-396
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• 2015

This study was carried out to investigate the action of phytohormones which influence the adventitious root formation of calli originating from the leaves of Persicaria perfoliata. The optimal medium condition for callus formation was ½-strength MS, 1% sucrose, and 4.5 μM 2,4-D. In order to determine which phytohormones had an effect on the adventitious root formation, the calluses were cultured in various media with different kinds of phytohormones. As a result, the medium with GA₃ or IAA was shown to induce root formation. To deeply investigate the effects of GA₃ and IAA, calli were cultured in 0.1, 1, and 10 mg/l levels of phytohormones. Numbers of roots formed per callus were 10.9, 14.2, 22.6 in GA₃, 5.8, 3.9, 1.1 in IAA, respectively. Therefore, the higher GA₃ or the lower IAA concentration, the more roots formed. To confirm this role of GA₃ we tested with inhibitors PBZ and NPA. GA₃ with PBZ resulted in reduction by 52.4~69.4% compared to GA₃ alone. In contrast, GA₃ with NPA resulted in an increase by -8~45.6% compared to GA₃ alone in root formation. Also, results were determined on the effect of GA₃ with other phytohormones on root formation. Kinetin, 2iP and ABA with GA₃ had a negative effect, but IAA with GA₃ showed a similar result to GA₃ alone. From these results we infer GA plays a key role and auxin has subsidiary activity on adventitious root formation. This is the first report that indicates GA₃ promotes adventitious root formation from calli in P. perfoliata.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.213-221
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• 2007

The treatment of radish cotyledons with a nitric oxide (NO)-releasing substance, sodium nitroprusside (SNP) resulted in an increased adventitious root development in a dose-dependent manner. However, this NO-mediated enhancement effect was reversed when either 0.5 mM EGTA (an extracellular $Ca^{2+}$ chelator) or 0.1 mM $LaCl_3$ (a calcium channel blocker) was applied with $50\;{\mu}M$ SNP. Our results also showed that guaiacol peroxidase (GPX) and syringaldazine peroxidase (SPX) activities, which are known to play a key role in rooting, were more largely increased during adventitious root induction in the cotyledons treated with SNP. However, the treatment of cotyledons with SNP plus $LaCl_3$ inhibited the SNP-induced increases in the activities of both GPX and SPX. Trifluoperazine (TFP), an antagonist of calmodulin (a specific calcium-binding protein), also delayed adventitious root formation and significantly reduced the root length and number of the SNP-treated cotyledons as well as the deactivation of GPX and SPX enzymes. In conclusion, our results suggest that calcium is involved in the NO response leading to induction of adventitious root through a regulation of GPX and SPX.

• Journal of Plant Biology
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• v.37 no.2
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• pp.195-201
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• 1994

In order to study the effects of polyamine inhibitors and polyamines on adventitious root formation, the correlation between adventitious root formation and carbohydrate metabolism was investigated in inoculated soybean (Glycine max L.) cotyledons at the concentration of $10^{-3}\;M$ methylglyoxal bis-(guanylhydrazone)[MGBG], and $10^{-3}\;M$ MGBG plus $10^{-5}\;M$ polyamines (putrescine, spermidine and spermine), respectively, for the adventitious root formation medium. The contents of starch, maltose and sucrose were lower in control and were higher in the treatments containing $10^{-3}\;M$ MGBG, and $10^{-3}\;M$ MGBG plus $10^{-5}\;M$ polyamines during culture. It was shown that the soluble sugar levels, except glucose, were higher than that of control in most of the treatments and the change in glucose contents tended to be similar to that control. The amylase activity increased in control and MGBG treatment, the maltase activity was higher in control and the invertase activity showed less substantial changes during culture.ulture.

• Korean Journal of Plant Tissue Culture
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• v.21 no.2
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• pp.105-110
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• 1994

In order to study on the effect of polyamine inhibitors and polyamines on adventitious root formation the correlation between adventitious root formation and polyamine levels were investigated in cultured soybean (Glycine max L.) cotyledons. Adventitious root formation was inhibited in medium containing 10$^{-4}$ -10/ sup -2/ M polyamine inhibition such as $\alpha$-difluoromethylornithine (DFMO), $\alpha$-difluoromethylarginine (DFMA), cyclohexylammonium sulfate (CHA), dicyclohexyl-amine (DCHA) and methylglyoxal-bis (guanylhydrazone) (MGBG). An inhibitory effects at 10$^{-3}$ M MCBG were much higher than other treatments. Treatment with 10$^{-3}$ M MGBG plus 10$^{-5}$ M spermine led to reversal of the effects of MGBG alone. The polyamine levels were sharply increased in the first few days in each treatnent compared to control. The remarkably increasing polyamine contents were observed in medium supplemented spermine.

• Korean Journal of Medicinal Crop Science
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• v.28 no.6
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• pp.445-454
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• 2020

Background: Culturing wild ginseng adventitious root using plant factory technology provides genetic safety and high productivity. This production technology is drawing attention in the fields of functional raw materials and product development. The cultivation method using elicitors is key technology for controlling biomass and increasing secondary metabolites. Methods and Results: Elicitor treatments using methyl jasmonate, pyruvic acid, squalene, β-sistosterol were performed to amplify total ginsenosides (Rb1, Rc, Rb2, Rb3, and Rd) of cultured wild ginseng adventitious root. Thereafter, fermentation and steaming processes were performed to convert total ginsenosides into minor molecular ginsenosides (Rg3, Rk1, and Rg5). The result indicated that methyl jasmonate minimizes the reduction in fresh weight of cultured wild ginseng adventitious root and maximizes total ginsenosides (sum of Rb1, Rc, Rb2, Rb3, and Rd). Ginsenoside conversion results showed a maximum degree of conversion of 131 mg/g. Conclusions: In this study, we demonstrated that the optimal elicitor treatment method increased the content of total ginsenosides, while the steaming and fermentation processing method increased the content of minor ginsenosides.

• Journal of the Korean Society of Tobacco Science
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• v.17 no.1
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• pp.33-40
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• 1995

Structures of the adventitious root meristem induced from callus culture of tobaco (Nicotiana tabacum cv. NC 82) leaves were investigated by light and transmission electron microscopy. Structural differences between in vitro root and callus cells were also examined by the microscopy. The submicroscopic features of the in vitro root cells were as follows. Intercellular spaces were not developed and nuclei with two nucleoli were observed occasionally. Plasmodesmate were found in groups or sing1y on transverse and longitudinal walls. Amyloplast solely filled with starch grains, with one to five electron - dense bands, was surrounded by single membrane. in the callus cells, vacuolization of central part in the cytoplasm having mitochondria with swollen cristae and starch grains like those of in vitro root cells was a distinct feature. Vesicles which were found between cell wall and plasma membrane may be arisen by a process of protoplasmic invagination. By comparing of ultrastructures between the cells of callus and in vitro roots we found that the distinct differences lied on thickened cell walls and hypertrophed vacuoles in the former, and less thickened cell walls and several small vacuoles in the later.

• Journal of Ginseng Research
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• v.31 no.3
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• pp.154-158
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• 2007

In order to increase the content of germanium in ginseng adventitious roots, the effects of chelating agents on germanium content and root growth were investigated in the submerged cultures of ginseng adventitious roots. Chelating agents such as citric acid, oxalic acid, phosphoric acid, EDTA (Ethylenediamine tetraacetic acid) or EGTA (Ethylene glycol-bis $({\beta}-aminoethylether)-tetraacetic$ acid) were administrated in the submerged culture of ginseng root containing 50 ppm $GeO_2$. After 6 weeks of cultivation, fresh weight, germanium and saponin contents in the roots were analyzed. Among chelating agents, addition of 1.0mM phosphoric acid was found to be best for germanium accumulation. Under this condition, germanium content increased 1.4 times as compared to that of the control. The germanium content in the adventitious roots also increased with addition of EDTA or EGTA, while they inhibited the growth of ginseng adventitious root. Citric and oxalic acids were not effective for increasing germanium content in adventitious roots. As the results, it suggests that the phosphoric acid can be proved as the optimal agent for the enhancement of germanium accumulation in ginseng adventitious roots. These results can be served as a guideline for the mass production of ginseng adventitious roots containing germanium by large-scale production.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.266.3-267
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• 2003

In order to induce production of methylnissolin, sterol biosyntheis inhibitor, in the adventitious root of Astragalus membranaceus the effect of methyl jasmonate(MeJ), a growth regulator of plant, was investigated. After treatment of MeJ (0$\mu\textrm{m}$, 10$\mu\textrm{m}$, 100$\mu\textrm{m}$) to the adventitious root which was harvested in the time interval of 0, 7, 14, 21, 28days and the fresh weight, dry weight and the contents methylnissolin was determined. (omitted)