• Plant Biotechnology Reports
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• 제2권2호
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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.

• 식물조직배양학회지
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• 제28권2호
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• pp.103-108
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• 2001

카사바 (Manihot esculenta Crantz cv. MColl 22)의 마디절편을 제아틴 0.01%가 첨가된 MS 기본배지에서 2주간 현탁배양하여 경엽부 길이가 1.5∼2.5 cm로 생장된 유식물의 부정근을(1) 1∼l.5 cm길이만 남기고 짧게 전정, (2) 1∼2 mm 이내로 제거, (3)뿌리 발생부 전체를 절단한 후 마사토를 담은 유리병에 이식하여 비무균조건에서 순화생장시켰다. 그 결과 이식 7∼10일경부터 각 조건에서 새 뿌리가 발생되기 시작하여 6주 후에는 조건에 따라 73∼93%의 생존율을 나타냈다. 기내 발생 뿌리 가운데 손상 없이 토양에 이식된 것은 그대로 생존을 계속하여 유식물 생장에 도움을 주었으며, 제거된 부정근은 이식 후 신장이 중단되고 새로 뿌리가 발생되었다. 그러나 간혹 전정되고 남은 부분에 측근이 발생된 것이 관찰되었다. 각 조건에서 새로 발생된 기외 뿌리의 수와 길이 및 경엽부 길이와 마디 수에서는 서로 유의성 있는 차이를 나타내지 않았으나, 생체중에서는 전정>제거>절단의 순으로 약간의 차이를 보였다. 미세증식된 카사바의 기내발생 부정근을 적절히 전정 또는 제거한 후 이식하여 순화시키는 것은, 작업을 간편하게 할 뿐만 아니라 유식물의 생존율이 높고 생장에도 지장이 거의 없으므로 경제적인 미세증식 방법으로 유용하게 이용될 수 있다.

• Mycobiology
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• 제30권2호
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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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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.151-162
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• 2001

산삼은 고유의 생약으로 민간 또는 한방에서 효능을 인정받아 왔으나 산삼의 희귀성으로 인하여 산삼에 대한 연구가 활발하지 못하였다. 따라서 본 실험은 식물 조직 배양 기술을 이용하여 산삼 부정근을 대량으로 배양하고 추출하여 화장품 원료로서의 적용 가능성을 연구하였다.

• 한국자원식물학회지
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• 제21권6호
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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.

• 한국자원식물학회지
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• 제22권6호
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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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• 제35권4호
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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.

• 한국식품과학회지
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• 제14권2호
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• pp.130-135
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• 1982

인삼중에 함유되어 있는 항산화물질의 본태를 파악하기 위하여 인삼의 각종 용매(petroleum ether, chleroform, chleroform-methanol, methanol 및 methanol-water)추출물들이 혼합 불포화지방산 메틸에스테르(MEUFA)의 자동산화에 미치는 영향을 in vitro에서 검토한 결과는 다음과 같다. 1. 인삼의 각종 용매 추출물들은 모두 항산화작용을 나타내어 MEUFA의 자동산화에 의한 지질 과산화물의 형성을 억제하였다. 그 중에서도 클로로포름 및 클로로포름-메탄올(2 : 1, v/v)추출물들이 보다 강한 항산화작용을 나타내었다. 2. Olcotte 등의 oven test에 의한 무게의 증가는 MEUFA의 자동산화에 의한 과산화물의 형성에 기인함이 확인되었다. 3. 인삼의 각 추출물들은 MEUFA의 지질 과산화물형성의 유도기간을 연장시켰다. 4. 각 추출물들의 항산화작용에 있어서 Olcott 등의 oven test에 의한 무게의 측정 Lea법과 Rhodan 철법에 의한 POV 측정 등의 결과에서 추정되는 작용은 상호간 잘 일치되었다. 이상의 결과로부터 인삼중에는 항산화물질들이 인삼성분으로서 함유되어 있음이 in vitro에서 확인되었다.

• Journal of Forest and Environmental Science
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• 제25권2호
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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%.

• 대한화장품학회지
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• 제27권2호
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• pp.45-56
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• 2001

산삼은 고유의 생약으로 민간 또는 한방에서 효능을 인정받아 왔으나 산삼의 희귀성으로 인하여 산삼에 대한 연구가 활발하지 못하였다. 따라서 본 실험은 식물 조직 배양 기술을 이용하여 산삼 부정근을 대량으로 배양하고 추출하여 화장품 원료로서의 적용 가능성을 연구하였다. 본 실험에서 사용한 산삼 부정근은 강원도 평창에서 채취한 110년생 천종삼으로 산삼으로부터 유래된 캘러스에서 부정근을 유도한 후 절취하여 생물 배양기에서 액체 현탁액으로 대량 배양하였다. 약 5주간의 배양기간을 거쳐 증식된 산삼 부정근을 세척하여 건조시킨 후 추출하여 산삼 부정근 추출물을 얻었다. 산삼 부정근 추출물의 미백 효과 측정을 위하여 tyrosinase 억제 실험과 DOPA 자동산화 그리고 B-16 melanoma cell를 이용한 미백 실험을 실시하였고 유해성 검증을 위해서 안전성 실험과 세포 독성 실험을 실시하였다. in vitro 상의 유해성 실험은 transformed mouse fibroblast L929를 배양하여 NR assay, MTT assay를, in vivo 상의 안전성 실험은 인체 첩포를 이용한 Patch test를 실시하여 피부 반응의 관찰을 통해서 자극 혹은 알레르기성 반응의 발생여부를 실시한 결과 무해하였으며 melanin 생성 억제 실험 결과 미백효과가 우수한 것으로 나타났다.