• 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%.

• 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.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2022년도 추계학술대회
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• pp.99-99
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• 2022

Artemisia fukudo is a biennial plant and has been reported to have anticancer, anti-melanogenesis, and anti-inflammatory effects. However, it is difficult to produce biomass from A. fukudo, so it is not used as a material for cosmetics or pharmaceuticals. In vitro culture can stably produce biomass throughout the year. In this study, the culture system for producing the highest biomass and bioactive substances was compared. Ex vitro plants were collected in Pyoseon-eup, Jeju island in May 2021, and in vitro culture was harvested after culturing for 8 weeks (plantlet) and 4 weeks (adventitious roots), respectively. After harvest, total polyphenol content (TPC), total flavonoid content (TFC), and DPPH scavenging activity were analyzed. In biomass production, adventitious roots (FW: 5.1 g·100 ml^-1, DW: 0.6 g·100 ml^-1) were about 4 times higher than that of plantlets (FW: 1.8 g·200 ml^-1, DW: 0.3 g·200 ml^-1). Both TPC and TFC were highest in ex vitro plants (9.2 ㎍·mL^-1, 31.6 ㎍·mL^-1), and were 3.0 times and 1.8 times higher than those of plantlets (3.0 ㎍·mL^-1, 17.8 ㎍·mL^-1), respectively. The IC₅₀ value of DPPH scavenging activity was also the best in ex vitro plants (69.8 ㎍·mL^-1), followed by root root (184.4 ㎍·mL^-1) and plants (325.3 ㎍·mL^-1) in that order. Through additional elicitor treatment, scale-up, and advanced compounds analysis such as HPLC, it can be used as an industrial material.

• 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.

• Journal of Plant Biotechnology
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• 제49권1호
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• pp.82-89
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• 2022

정금나무(Vaccinium oldhamii)의 정아를 포함한 줄기를 재료로 다신초 유도 및 기외발근을 통해 완전한 식물체를 효과적으로 재분화시킬 수 있었다. 정아를 포함한 줄기로부터 다신초 유도는 2.0 mg/L 2-iP에서 유도율 100.0%, 절편체 당 다신초 유도수 7.4개로 나타났으며, 특히 신초 길이는 평균 51.7 mm로 다른 사이토키닌(zeatin, BA 또는 TDZ) 처리구에 비해 가장 높게 나타났다. 증식된 줄기로부터 기내발근은 0.5 mg/L IBA 첨가에서 절편체 당 2.4개의 뿌리를 생산하였고, 뿌리 길이는 평균 17.6 mm로 가장 우수한 결과를 보였다. 상토를 이용한 기외발근의 경우 1.0 mg/L IBA와 탈크(talc) 혼합용액 처리 시 평균 발근율은 80.0%로 기내발근에 비해 더 양호한 것으로 나타났다.

• Archives of Pharmacal Research
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• 제14권4호
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• pp.352-356
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• 1991

The anti-platelet activity of ilexoside D isolated from the roots of Ilex pubescens Hook. et Arn. was investigated in in vitro and ex vivo models of platelet aggregation induced by ADP, thrombin or collagen in rats. In vitro ilexoside D inhibited more effectively platelet aggregation induced by ADP and thrombin than by collagen as compared with aspirin. Ex vivo ilexoside D also inhibited platelet aggregation induced by ADP and collagen, but not by thrombin, and the inhibitory action of ilexoside D was more effective than that of aspirin. However, in vitro ilexoside D inhibited very poorly the generation of malonyldialdehyde, which is known to be concomitantly released with thromboxane $A_2$ during platelet aggregation. These results suggest that the anti-platelet activity of ilexoside D may not be responsible for prostaglandin synthesis in platelets.

• Journal of Forest and Environmental Science
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• 제37권4호
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• pp.304-308
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• 2021

Iris koreana (Iridaceae) is an endangered plant native to Korea. In order to develop an in vitro propagation method, we investigated the effect of 2,4-dichlorophenoxy acetic acid (2,4-D) and a-naphthalene acetic acid (NAA) on callus induction in different I. koreana tissues. In addition, we also investigated the effect of 2,4-D and Benzyl aminopurine (BA) treatments on adventitious shoot induction in viable calli and the effect of indole-3-butyric acid (IBA) on root formation in viable shoots. We found that callus production was highest with 1.0 mg/L NAA (94.4% cultured rhizome explants), and adding low concentrations of 2,4-D to BA containing media significantly increased the frequency of shoot primordial formation. The best rooting results were obtained with 1.0 mg/L IBA, on which 98% of regenerated shoots developed roots and produced an average of 7.4 roots within 45 days. This in vitro propagation protocol will be useful for conservation, as well as for mass propagation.

• 원예과학기술지
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• 제32권5호
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• pp.694-701
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• 2014

Allium hookeri L. (Alliaceae family) is an important ethnomedicinal plant native to the Himalayan region of Asia. The aim of this research was to establish a high-frequency plant regeneration system for in vitro propagation of A. hookeri. Among the tissue types examined, receptacle explants derived from immature flower buds showed the highest regeneration rate of shoots ($93.33{\pm}4.63%$), roots ($76.67{\pm}7.85%$), and calli ($80.00{\pm}7.43%$) when cultured on Gamborg B5 (B5) medium containing $10{\mu}M$ 6-benzylaminopurine (BA) + $1{\mu}M$ naphthalene acetic acid (NAA), $0.5{\mu}M$ BA + $5{\mu}M$ NAA, and $1{\mu}M$ BA + $10{\mu}M$ NAA, respectively. Shoot multiplication was superior when cultured in liquid rather than on solid medium and relatively high concentrations of BA, ranging from 5 to $10{\mu}M$. Efficient bulblet formation following root induction from shoot clumps was achieved with culture in liquid B5 medium containing 7% (w/v) sucrose. Regenerated bulblets were successfully acclimatized to ex vitro conditions with a greater than 95% survival rate. By this method, a maximum of 62 plantlets per receptacle could be propagated within 9 weeks of initial culture. The in vitro propagation system established in this study will promote A. hookeri biotechnology, including large-scale production of healthy and aseptic clones, preserving parental genotypes with desirable traits, and genetic manipulation to enhance medicinal value.

• 한국산림과학회지
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• 제96권2호
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• pp.178-182
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• 2007

Rauvolfia serpentina (L.) Benth. ex Kurz is a medicinal plant and an endangered tropical rainforest plant species. Since the field cultivation that aims to fulfill the industrial needs is never accomplished, tissue culture appears to be the most feasible way to improve the quality and quantity of R. serpentina. This experiment used two kinds of explants (roots and shoots) to induce optimal root formation in different combinations of auxin and photoperiod. Each explants exhibited different responses on given treatments. Differentiated root could be produced from explants cultured in IBA 20 mg/L with and without light. The highest number of roots, root length and root weight induced from shoot explants were effective on MS medium containing IBA 20 mg/L and incubated under dark condition, while highest total weight (callus and root) from root explants cultured on MS medium supplemented 10 mg/L IBA and 10 mg/L NAA and incubated under day length (11/13 hr). The root induced from shoot explants produced the highest major alkaloid content. The highest content of ajmaline (2.17 ppm fresh weight) and reserpine (1.30 ppm fresh weight) were observed in shoot explants cultured in MS medium containing combination of IBA 10 mg/L and NAA 10 mg/L and incubated under dark condition, yohimbine (1.47 ppm fresh weight) was in the shoot explants cultured in MS medium containing NAA 20 mg/L and incubated under day length, while serpentine was absent.

• Journal of Plant Biotechnology
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• 제39권2호
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• pp.114-120
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• 2012

본 실험은 약리적 효능이 뛰어난 홍경천의 기관 (액아가 포함된 줄기 조직)배양을 통한 대량증식 방법의 일환으로 액아배양을 통한 다신초를 유도, 증식조건에 관해 BA와 GA3의 영향을 조사하였고 신초증식에 필요한 적정 sucrose의 농도를 확인하였다. 그 후 적절한 기내 발근 및 토양순화 조건을 탐색하여 최적의 홍경천의 생산 조건을 확립하였다. 다신초의 유도 및 증식은 $GA_3$와 BA를 혼합 처리가 효과적이었다. 신초증식에 효과적인 sucrose의 농도는 50 g/L가 첨가된 배지였으며, 평균 7 cm 이상의 신장을 보였다. 기내 발근은 옥신류 식물생장조절물질 무첨가 배지에서 가장 높은 발근길이와 개수를 나타냈다. 기내발근된 유식물체의 토양순화는 모래상을 제외하고는 모두 양호한 생장을 보였고, 특히 모래, 피트모스, 펄라이트가 모두 첨가된 배합토에서 가장 양호한 생장을 보였다. 이상의 결과는 수년전 국내에 도입하려다 실패한 약용식물인 홍경천의 기내배양을 통한 재배의 가능성을 제시하는 것이고 더 나아가 다양한 생물소재의 활용 방안의 기초재료로 활용될 것으로 보여진다.