• Plant Biotechnology Reports
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• v.2 no.2
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.103-108
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• 2001

The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

• Mycobiology
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• v.30 no.2
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• pp.70-75
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• 2002

The mycelia were directly isolated from eight species of fungal basidiocarps, confirmed to the ectomycorrhiza in the roots from the fields(forestry); Suillus bovinus, Paxillus involutus, Lactarius hysginus, Russula fragilis, Lepista nuda, Lyophyllum shimeji, Tricholoma matsutake, and Russula integra. The mycelia were pure-cultured with several transferring in various agars, and inoculated to the roots of pine(Pinus densiflora) seedling by in vitro method. After ten months growth under artificially aseptic conditions, all pine seedlings inoculated were stimulated at the growth-height, whereas those not inoculated were nearly dead. Also, the ramifications of ectomycorrhizal pine roots formed in the synthetic in vitro systems and were various according to the different mycelia. Synthesis of ectomycorrhiza were clearly confirmed in ten months growth, but not distinguished at this moment. It was clearly proved that the mycelia isolated caused the ectomycorrhizae in the roots of pine seedlings.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.151-162
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• 2001

Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extrac. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were patch tested in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynnthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively

• Korean Journal of Plant Resources
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• v.21 no.6
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• pp.444-448
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• 2008

An adventitious root formation protocol from Smilax china L. was established for in vitro production of dioscin, a steroidal saponin having various bioactivities such as anticancer, antifungal, antiviral, and antiobesity. Optimal medium for root initiation from leaf explant was MS medium containing $30\;g{\cdot}L^{-1}$ of sucrose supplemented with $1.0\;mg{\cdot}L^{-1}$ kinetin + $2.0\;mg{\cdot}L^{-1}$ NAA. The induction of adventitious roots from in vitro initiated root segments was most favorable to MS liquid medium with $0.1\;mg{\cdot}L^{-1}$ kinetin + $2.0\;mg{\cdot}L^{-1}$ NAA. Among the 20 different adventitious roots originated from different plants, strain No. 10 was selected based on production ability of dioscin, and its stability through the successive suspension culture. The maximum growth stage of adventitious roots was noticed at 5 weeks after subculture while that of dioscin production in the adventitious root was at 7 weeks after subculture in suspension culture system. These results provide that suspension culture of adventitious roots of Smilax china L. have a potential for in vitro mass production of dioscin.

• Korean Journal of Plant Resources
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• v.22 no.6
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• pp.584-591
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• 2009

Adventitious root culture was established in the Jawahar variety of Withania somnifera using MS basal medium supplemented with 0.5 (mg/l) IAA and 2.0 (mg/l) IBA. Root tips from germinated seedlings, MS0 maintained plants and adventitious roots were maintained in suspension medium (1/2 MS basal medium supplemented with 3% sucrose) for a period of 1 to 6 months. The weight gain in roots was noted and the withanolides were extracted from the dry roots using solvents petroleum ether, 50% ethanol and chloroform. The withanolides in the chloroform fractions of all root samples analyzed were compared using thin layer chromatographic analysis. Withanolide content in adventitious root sample was found to be superior compared to other roots at any given point of time during the 6month growth period.HPLC analysis of in vitro adventitious roots showed the presence of a new compound.

• Journal of Ginseng Research
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• v.35 no.4
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• pp.442-448
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• 2011

We previously reported that in vitro anti-platelet activity of tissue-cultured mountain ginseng (TCMG) ethanol extracts show improved efficacy when compared with commercial ginseng products such as Korean red ginseng and Panax ginseng. However, information on the anti-platelet activity of the ethyl acetate fraction from TCMG adventitious roots is limited. Therefore, in this study, we further investigated the effects of an ethyl acetate extract of TCMG (EA-TCMG) adventitious roots on in vitro antiplatelet activity in whole human blood and its effect on peripheral blood flow in mice. We found that EA-TCMG inhibited platelet aggregation with $IC_{50}$ values of 271, 180, and 147 ${\mu}g$/mL induced by collagen, adenosine-5'-diphosphate, and arachidonic acid, respectively. Among the three agonists used, thromboxane $A_2$ formation induced by arachidonic acid was markedly suppressed. Furthermore, EA-TCMG improved the peripheral circulatory disturbance by improving vascular blood flow. In conclusion, these results suggest that ethyl acetate extracts from TCMG adventitious roots might inhibit vascular platelet aggregation and thrombus formation.

• Korean Journal of Food Science and Technology
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• v.14 no.2
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• pp.130-135
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• 1982

The antioxidative action of petroleum ether, chloroform, chloroform-methamol (2 : 1, v/v), methanol and methanol-water extracts of Panax ginseng roots in the autoxidation of mixed methyl esters of unsaturated fatty acids (MEUFA) was investigated in vitro. All of the extracts of Panax ginseng roots possessed antioxidative activity and inhibited the formation of hydroperoxides in the autoxidation of mixed methyl esters of unsaturated fatty acids (MEUEA). The induction period in the autoxidation of MEUFA was extended by the addition of each extract. The antioxidative activity was more prominent with chloroform and chloroform-methanol (2 : 1, v/v) extracts than with other extracts. The antioxidative activity of each extract estimated by Olcott's oven test and the measurement of peroxide value (POV) showed a similar tendency. From the results obtained, it was conclude that extracts of Panax ginseng roots had remarkable antioxidative activity in the autoxidation of MEUFA in vitro.

• Journal of Forest and Environmental Science
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• v.25 no.2
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• pp.119-126
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• 2009

An efficient in vitro plant regeneration system was developed through shoot proliferation from axillary buds of Tectona grandis (L.f), APNBV-1 (Andhra Pradesh North Badrachalam Venkatapuram-1) clone. Multiple shoots of high quality were produced in vitro from axillary bud explants. An average of 4.39 shoots/explant were obtained on Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) benzyl amino purine (BA), kinetin (KN), indole acetic acid (IAA), gibberillic acid ($GA_3$), growth adjuvants casein hydrolysate (CH), adenine sulphate (Ads) and antioxidants ascorbic acid, polyvinyl pyrrollidine (PVP). Eighty five percent of rooting was observed in ex vitro rooting media containing IBA and vermiculite. In ex vitro rooting, single shoots with 2 to 3 nodes were subjected to IBA of different concentrations at different periods of time intervals. Direct rooting in vermiculite at 500 ppm concentration of IBA resulted in 4.3 number of roots with 2 cm length. Minimum response of rooting and length of roots were recorded at 100 ppm concentration of IBA. Planlets were transferred to plastic bags for short acclimatization stage in green house where they survived at 95%.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.2
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• pp.45-56
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• 2001

Korea mountain ginseng known as oriental miracle drug is an important medicinal plant. The effect of mountain ginseng adventitious roots extract has been described. The valuable root of mountain ginseng contained several kinds of ginsenosides that have been confirmed to have many active functions for the human body. However, the study of mountain ginseng has a limit because the price of wild ginseng is very expensive and rare. The mountain ginseng adventitious roots were derived from mountain ginseng callus that were induced from mountain ginseng roots. Adventitious roots were separated from callus and grown in solid media(Murachige and stoog media). It was cultured in a 20L bioreactor. After culturing for 40days, adventitious roots were harvested. Afterwards the harvested mountain ginseng adventitious roots were dryed and extracted. We examined the effect on melanogenesis of mountain ginseng adventitious roots extract. Here, we report the inhibitory effect of melanin biosynthesis on the adventitious roots extract of In vitro test. Also, we assessed the safety of adventitious roots extract. In vitro, cytotoxicity of adventitious roots extract was assessed in mouse fibroblast using two method: The neutral red uptake assay and the MTT assay. In vivo, the allergic and irritant were Patch teated in 30 patients. Consequently, extract of mountain ginseng adventitious roots have inhibitory effect on melanin biosynthesis in B-16 melanoma cell test, tyrosinase inhibitory test and DOPA auto-oxidation test. There were decreased 86%(0.5% concentration), 45%(1% concentration) and 61%(1% concentration), respectively.