• Title/Summary/Keyword: adventitious root

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Adventitious Root Cultures of Panax ginseng C.V. Meyer and Ginsenoside Production through Large-Scale Bioreactor System

  • Hahn, Eun-Joo;Kim, Yun-Soo;Yu, Kee-Won;Jeong, Cheol-Seung;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.5 no.1
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    • pp.1-6
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    • 2003
  • The adventitious root of Panax ginseng C.A. Meyer is regarded as an efficient alternative to cell culture or hairy root culture for biomass production due to its fast growth and stable metabolite production. To determine optimal culture conditions for the bioreactor culture of ginseng roots, experiments have been conducted on physical and chemical factors such as bioreactor type, dissolved oxygen, gas supply, aeration, medium type, macro- and micro-elements, medium supplement during culture period, sucrose concentration, osmotic agents, medium pH and light. Elicitation is a key step to increase ginsenoside accumulation in the adventitious roots but biomass growth is severely inhibited by elicitor treatment. To obtain high ginsenoside content with avoiding biomass decrease, we applied two-stage bioreactor culture system. Ginseng adventitious roots were cultured for 40 days to maximize biomass increase followed by elicitation for 7 days to enhance ginsenoside accumulation. We also experimented on types and concentrations of jasmonate to determine optimal elicitation methods. In this paper, we discussed several factors affecting the root propagation and ginsenoside accumulation. Based on the results obtained from previous experiments we have established large-scale bioreactor system (1 ton-10 ton) for the efficient production of ginseng adventitious roots and bioactive compounds including ginsenoside. Still, experiments are on going in our laboratory to determine other bioactive compounds having effects on diet, high blood pressure, DPPH elimination and increasing memories.

Effects of Inverted Incubation on Adventitious Root Formation in Epicotyl Cuttings of Vigna angularis Owhiet Ohashi (팥의 유경절편에서 부정근형성에대한 도립배양의 효과)

  • Kang, Byung-Sook;Cho, Duck-Yee;Kim, Young-Soon;Soh, Woong-Young
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.75-80
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    • 1998
  • Disbudded epicotyl cuttings from light-grown 6-day-old seedings of Vigna angularis Owhi et Ohashi were preincubated in $2\;\times\;10^{-4}M$ IAA solution for 48 hr to promote adventitious root formation in upright or inverted direction and then incubated in upright direction for 96 hr. Adventitious root formation occurred only at the morphological base of the cuttings which were preincubated in upright direction, while at the both ends in inverted direction. IAA treatment enhanced the adventitious root formation in all cuttings regardless of their orientation during preincubation. To elucidate localized root development, the activity of enzymes involved in root initiation and development was measured 24 hr, 48 hr, and 148 hr after epicotyl incubation. IAA oxidase, peroxidase and catalase were assayed in the apical, middle and basal segment of the epicotyls, and their fresh weight and length were measured. Elongation occurred the most in the upper segment of the epicotyl while fresh weight gain was the most in the basal segment. At root initiation phase, 24 hr after incubation IAA peroxidase and catalase activities appeared high at rooting zone while IAA oxidase activity was low at both ends, IAA oxidase and peroxidase activities declined at the rooting zone during the adventitious root formation at 48 ht. Inversion of cuttings during preincubation caused a chrange of enzyme activities along their epicotyl cuttings. Only peroxidase activity showed a high correlation with root initiation. Therefore, the biochemical change is highly correlated with change in IAA level in the rooting zone of the epicotyl, resulting in root formation in unusual rooting zone of epicotyl.

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Growth, secondary metabolite production and antioxidant enzyme response of Morinda citrifolia adventitious root as affected by auxin and cytokinin

  • Baque, Md. Abdullahil;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.109-116
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    • 2010
  • Morinda citrifolia adventitious roots were cultured in shake flasks using Murashige and Skoog medium with different types and concentrations of auxin and cytokinin. Root (fresh weight and dry weight) accumulation was enhanced at 5 $mg\;l^{-1}$ indole butyric acid (IBA) and at 7 and 9 $mg\;l^{-1}$ naphthalene acetic acid (NAA). On the other hand, 9 $mg\;l^{-1}$ NAA decreased the anthraquinone, phenolic and flavonoid contents more severely than 9 $mg\;l^{-1}$ IBA. When adventitious roots were treated with kinetin (0.1, 0.3 and 0.5 $mg\;l^{-1}$) and thidiazuron (TDZ; 0.1, 0.3 and 0.5 $mg\;l^{-1}$) in combination with 5 $mg\;l^{-1}$ IBA, fresh weight and dry weight decreased but secondary metabolite content increased. The secondary metabolite content (including 1,1-diphenyl-2-picrylhydrazyl activity) increased more in TDZ-treated than in kinetin-treated roots. Antioxidative enzymes such as catalase (CAT) and guaiacol peroxidase (G-POD), which play important roles in plant defense, also increased. A strong decrease in ascorbate peroxidase activity resulted in a high accumulation of hydrogen peroxide. This indicates that adventitious roots can grow under stress conditions with induced CAT and G-POD activities and higher accumulations of secondary metabolites. These results suggest that 5 $mg\;l^{-1}$ IBA supplementation is useful for growth and secondary metabolite production in adventitious roots of M. citrifolia.

Determination Times for Induction of Adventitious Shoots, Roots, Trichomes, and Calli from Segments of Arabidopsis thaliana by NAA and BA (애기장대(Arabidopsis thaliana)잎 절편에서 NAA와 BA에 의한 신초, 부정근, 모용 및 캘러스 형성 결정 시기)

  • Kim, Song-Lim;Han, Tae-Jin
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.161-165
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    • 2003
  • The effect of NAA and Benzyladenine(BA) for determination times on the formation of adventitious shoots, roots trichmoes and calli in MS basal medium was investigated in leaf segments from ecotype 'Nosses' of Arabidopsis thalliana. Adventitious shoots, roots, trichomes and calli were formed fromed from leaf segments in a wede range of NAA and BA. The optimal combination of hormones for adventitious shoots formation, 20mg/L NAA for trichome formation, 100mg/L for callus formation. Inductive times for formation of adventitious shoots, roots, trichomes and calli were determined at 14, 4, 6 and 18 days respectively by periodical transfer of leaf segments from hormines containing media to hormone free medium.

Induction and in vitro Proliferation of Adventitious Roots in Phyllanthus urinaria (여우구슬(Phyllanthus urinaria)의 부정근 유도 및 기내증식조건)

  • Bae, Kee-Hwa;Yun, Pil-Yong;Choi, Yong-Eui
    • Korean Journal of Plant Resources
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    • v.22 no.5
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    • pp.454-460
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    • 2009
  • Phyllanthus urinaria was an important species in Korea and distributed in all around of Korea. The roots and stems of this plant have been used for natural medicine for the treatment of diabetes, the hepatitis B virus and disturbances of the kidney and urinary bladder. Production of adventitious roots in P. urinaria by in vitro cultures could be used as alternatives materials. Shoot and root segments from P. urinaria seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the shoot part. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximun induction of adventitious root was obtained on medium with 0.5 mg/L IBA. In liquid culture, growth of root was best on medium supplemented with 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 0.5 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. These results revealed the first attempt for the production of adventitious roots in P. urinaria.

Plant Regeneration via Adventitious Shoot Formation from Hypocotyl Explants of Groundcherry (Physalis angulata L.) (땅꽈리(Physalis angulata L.) 하배축 절편으로부터 신초 형성을 통한 식물체 재분화)

  • Koh, Seok Chan
    • Korean Journal of Plant Resources
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    • v.35 no.4
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    • pp.502-507
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    • 2022
  • In the present study, plant regeneration through adventitious shoot formation from hypocotyl segments of in vitro seedlings of groundcherry (Physalis angulata L.) was investigated to determine the optimum culture conditions for highly efficient regeneration of the species. Adventitious shoots in hypocotyl segments were efficiently induced on MS media with low concentrations of BAP, specifically, with 0.5-1.0 mg/L BAP singly or in combination with 0.1-0.5 mg/L NAA. The 1.0 mg/L BAP single treatment was most effective for forming multiple adventitious shoots. When the induced shoots were transferred to the root induction media, low concentrations of NAA, IBA, and IAA enhanced the development of adventitious roots from adventitious shoots, suggesting that low concentrations of auxins were optimal for producing regenerated plantlets. The number of roots per shoot was large (> 2.0), and the root length exceeded 8.0 cm. In particular, the development and the overall shape of the roots were ideal. Furthermore, the number and length of shoots exceeded 2 and 6.0 cm, respectively. When the regenerated plantlets were transferred to compost soil, the root and shoot systems had developed well to the point that all of the regenerated plantlets acclimated successfully, resulting in normal morphology and growth characteristics, similar to those of the mother plant. Therefore, plant regeneration via adventitious shoot formation is expected to be one of the main methods for producing groundcherry on a large scale for a stable supply of the raw materials.

Regeneration of adventitious root from Calystegia soldanella L. in Jeju island and mass proliferation method using bioreactor system (제주지역 갯메꽃(Calystegia soldanella L.) 유래 부정근 재분화 및 생물반응기 시스템 이용 대량증식법)

  • Jong-Du Lee;Eunbi Jang;Weon-Jong Yoon;Yong-Hwan Jung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.37-37
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    • 2021
  • Calystegia soldanella L. is a perennial herbaceous halophyte belonging to the convolvulaceae family, which mainly grows in coastal sand dunes in Korea. Shoots and rhizomes are edible, and roots called 'Hyoseon Chogeun' are known to have medicinal effects such as antipyretic, sterilization, and diuretic. In addition, physiological activities of antioxidant, anti-inflammatory, antiviral, antifungal and PTP-1B (protein tyrosine phosphate-1B) inhibition have been reported. In this study, in vitro induction cell lines of C. soldanella L. collected from the coastal sand dunes in Jeju island was redifferentiated into adventitious roots that can be used as medicinal resources. Also the biomass of mass-proliferated adventitious roots using a bioreactor were evaluated. Plants of C. soldanella L. were collected from the crevice of the seashore in the coastal area of Taeheung 2-ri, Namwon-eup, Seogwipo-si. Then, it was separated into leaves, stems, rhizomes, and roots, and surface sterilized with 70% ethyl alcohol and 2% NaOCl (sodium hypochlorite). After washing with sterilized water, each organ section was cultured in Hormone-free MS medium (Murashige & Skoog Medium). As a result, the induction response rates were evaluated at 85% and 55%, respectively, in terms of callus formation and shoot generation in the rhizome segment. In the case of the adventitious roots morphological characteristics induced by single-use treatment of auxin-based plant growth regulators IBA and NAA from redifferentiated shoots were compared. Most efficient adventitious root culture method as a rooting rate, number, length, and biomass proliferation in the bioreactor system was confirmed when treated by culturing in MS salts, Sucrose 30 g·L-1, and IBA 1mg·L-1 for 4 weeks. In this study, the medium composition and culture period were confirmed using a bioreactor system to mass-proliferate adventitious roots derived from C. soldanella L. in Jeju island. Also this adventitious root line developed a new medicinal material could increase value of the bio-industry ingredient through quantitative and qualitative screening of phyto-bioactive compounds.

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Adventitious Shoot Formation and Plant Regeneration from Explants of Solanum nigrum L. (까마중(Solanum nigrum L.)의 유식물 절편체에서 부정아 형성 및 식물체 재분화)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.277-284
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    • 2012
  • In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

Optimal conditions for adventitious root organogenesis from peony root explant callus cultures (작약(Paeonia lactiflora Pall.) 뿌리절편 유래 캘러스 배양으로부터 부정근발생을 위한 최적 배양조건)

  • Lee, Young Jin;Choi, Myung Suk;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.207-212
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    • 2022
  • The optimal culture conditions for root organogenesis from the callus of peonies (Paeonia lactiflora Pall.) were investigated. Root explants with vascular bundles were cultured in Murashige and Skoog (MS) medium combined with 0.5-4.0 mg/L auxins (indole acetic acid [IAA], naphthalene acetic acid [NAA], indolebutyric acid [IBA], and 2,4-dichlorophenoxyacetic acid [2,4-D]) and 0.0-2.0 mg/L cytokinins (kinetin, zeatin, and benzylaminopurine [BAP]) to induce callus formation. The callus was then cultured in MS medium combined with three concentrations (0.1, 0.5, and 1.0 mg/L) of IAA, NAA, IBA, kinetin, zeatin, and BAP in the dark for 6 weeks. Based on the results, the effects of dark and light conditions on the callus cultured in MS medium with combinations of 0.1-1.0 mg/L IBA and zeatin for 6 weeks were studied. Callus formation was most effective (>+++) in the medium with a combination of 1.0 mg/L NAA and 1.0 mg/L zeatin. A high number of long adventitious roots were observed in the mediums with 0.1 mg/L IBA (6.66 and 4.82 cm) and 0.5 mg/L zeatin (2.32 and 0.72 cm) among auxins and cytokinins, respectively. The highest number (14.06) of adventitious roots were formed from the callus cultured in light in the MS medium combined with 0.1 mg/L IBA and 0.5 mg/L zeatin. This same medium induced the formation of the longest adventitious root (5.45 cm) in the dark. Thus, optimization of in vitro culture conditions may be possible for the mass propagation of adventitious roots in peonies.

산삼의 배양 및 그 응용에 관한 연구

  • Sin, Mi-Hui
    • 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

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