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Inhibition of Adventitious Root Growth in Boron-Deficient or Aluminum-Stressed Sunflower Cuttings

  • Hong, Jung-Hee;Go, Eun-Jung;Kim, Tae-Yun
    • Journal of Environmental Science International
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    • v.12 no.11
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    • pp.1189-1196
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    • 2003
  • The effect of boron and aluminum on the development of adventitious roots was studied in sunflower cuttings. Three-day-old seedlings were de-rooted and grown in nutrient solutions with or without boron and supplemented with different concentrations (from 50 to 700 ${\mu}$M) of aluminum. The number and length of the adventitious roots and proline content in adventitious roots in response to insufficient boron and aluminum stress were determined periodically. The micronutrient boron caused the development of numerous roots in the lower parts of the hypocotyl. A dose-response of boron-induced rooting yielded an optimum concentration of 0.1 mM boron. In the absence of boron, in the majority of the adventitious roots, a significant inhibition was observed with or without aluminum, indicating that the most apparent symptom of boron deficiency is the cessation of root growth. Increasing concentrations of aluminum caused progressive inhibition of growth and rooting of the hypocotyls, and a parallel increase in proline levels of adventitious roots. Supplemental boron ameliorated the inhibitory effect of aluminum, suggesting that aluminum could inhibit root growth by inducing boron deficiency. Ascorbate added to medium in the absence of boron improved root growth and induced a significant decrease in proline levels. These findings suggest that adventitious root growth inhibition resulting from either boron deficiency or aluminum toxicity may be a result of impaired ascorbate metabolism.

Adventitious root culture for secondary metabolite production in medicinal plants: A Review

  • Rahmat, Endang;Kang, Youngmin
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.143-157
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    • 2019
  • Medicinal plants are high-value natural resources that have been used as precautionary drugs by many people globally. The increasing global demand for bioactive compounds from medicinal plants has led to the overexploitation of many valuable species. One widely used approach to overcome this problem is the use of adventitious root cultures as a propagation strategy. This review examines the scientific research published globally on the application of adventitious root cultures for many medicinal plants. Adventitious roots generated under aseptic environments in suitable phytohormone-augmented medium exhibit high growth rates and production of important secondary metabolites. Parameters such as medium properties and composition, growth hormone type, and elicitation strategies for in vitro grown adventitious roots of medicinal plants, are the main topics discussed in this review. We also examine current developments in bioreactor system cultivation for plant bioactive compounds using adventitious root cultures, a technology with possible commercial applications, via several studies on adventitious root culture of medicinal plants in which bioreactor systems play a role. In conclusion, the development of adventitious root cultures for medicinal plants is highly useful because of their capability for vegetative propagation and germplasm preservation.

Optimization of Extraction Condition of Methyl Jasmonate-treated Wild Ginseng Adventitious Root Cultures using Response Surface Methodology

  • Liu, Qing;Jo, Yang Hee;Ahn, Jong Hoon;Kim, Seon Beom;Paek, Kee-Yoeup;Hwang, Bang Yeon;Park, So-Young;Lee, Mi Kyeong
    • Natural Product Sciences
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    • v.24 no.2
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    • pp.103-108
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    • 2018
  • The usage of wild ginseng (Panax ginseng C.A. Meyer) has been limited due to short supply and high price. Therefore, sufficient production as well as efficient extraction of mountain ginseng are required for the development as products. In this study, wild ginseng adventitious root cultures were prepared for efficient production with advantages of fast growth and stable production. Treatment of methyl jasmonate (MJ) to wild ginseng adventitious root cultures increased the extraction yield and antioxidative activity. Further investigation on effect of extraction conditions suggested the importance of ethanol concentration on antioxidative activity and extraction yield of MJ-treated wild ginseng adventitious root cultures. Optimized extraction condition of MJ-treated wild ginseng adventitious root cultures for maximum extraction yield and antioxidative activity was determined using response surface methodology with three-level-three-factor Box-Behnken design (BBD). Extraction of 1 g MJ-treated wild ginseng adventitious root culture with 30 ml of 9% ethanol at $30^{\circ}C$ produced 310.2 mg extract with 71.0% antioxidative activity at $100{\mu}g/ml$. Taken together, MJ-treated wild ginseng adventitious root culture is valuable source for wild ginseng usage and optimized extraction condition can be used for the development of functional products or folk remedies.

Involvement of nitric oxide-induced NADPH oxidase in adventitious root growth and antioxidant defense in Panax ginseng

  • Tewari, Rajesh Kumar;Kim, Soohyun;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Plant Biotechnology Reports
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    • v.2 no.2
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    • pp.113-122
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    • 2008
  • Nitric oxide (NO) affects the growth and development of plants and also affects plant responses to various stresses. Because NO induces root differentiation, we examined whether or not it is involved in increased ROS generation. Treatments with sodium nitroprusside (SNP), an NO donor, 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger, and $N{\omega}-nitro-{\text\tiny{L}}-arginine$ methyl ester hydrochloride (${\text\tiny{L}}-NAME$), an NO synthase (NOS) inhibitor, revealed that NO is involved in the adventitious root growth of mountain ginseng. Supply of an NO donor, SNP, activates NADPH oxidase activity, resulting in increased generation of $O_2{^{{\cdot}-}}$, which subsequently induces growth of adventitious roots. Moreover, treatment with diphenyliodonium chloride (DPI), an NADPH oxidase inhibitor, individually or with SNP, inhibited root growth, NADPH oxidase activity, and $O_2{^{{\cdot}-}}$ anion generation. Supply of the NO donor, SNP, did not induce any notable isoforms of enzymes; it did, however, increase the activity of pre-existing bands of NADPH oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase, and glutathione reductase. Enhanced activity of antioxidant enzymes induced by SNP supply seems to be responsible for a low level of $H_2O_2$ in the adventitious roots of mountain ginseng. It was therefore concluded that NO-induced generation of $O_2{^{{\cdot}-}}$ by NADPH oxidase seems to have a role in adventitious root growth of mountain ginseng. The possible mechanism of NO involvement in $O_2{^{{\cdot}-}}$ generation through NADPH oxidase and subsequent root growth is discussed.

Effects of Auxin-induced Ethylene on Growth and Development of Adventitious Roots of Panax ginseng C.A. Meyer (IBA와 NAA 처리에 의해 생성된 Ethylene이 인삼(Panax ginseng C.A. Meyer) 부정근의 생장과 발달에 미치는 영향)

  • Kim, Yun-Soo;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.173-177
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    • 2003
  • The effect of IBA and NAA on adventitious root cultures of Panax ginseng C.A. Mater were investigated. Results indicated differences in growth and development of the roots according to 5mg/L IBA and 2mg/L NAA. IBA resulted in a normal root development and a higher growth compared to NAA. The roots formed on NAA-containing media were shorter and thicker than those in IBA, showing a hypertrophy of the root tip. NAA induced more than 1.6 times higher ethylene production compared to IBA, which caused inhibition of the root growth. Under the ventilation, in the other hand, on difference was observed in ethylene concentration and the root growth between IBA and NAA treatments. Under ventilation ethylene production was not detected until 10 days of culture, while detected from the initial stage under on ventilation. The results suggested the importance of ventilation during the culture for the growth and development of ginseng adventitious roots.

A Simple and Rapid Method for Functional Analysis of Plant Growth-promoting Rhizobacteria Using the Development of Cucumber Adventitious Root System

  • Bae, Yeoung-Seuk;Park, Kyung-Seok;Lee, Young-Gee;Choi, Ok-Hee
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.223-225
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    • 2007
  • Many plant growth-promoting rhizobacteria (PGPRs) have been known for beneficial effects on plants including biological control of soilborne pathogens, induced systemic resistance to plant pathogens, phytohormone production, and improvement of nutrient and water uptake of plants. We developed a simple and rapid method for screening potential PGPR, especially phytohormone producing rhizobacteria, or for analyzing their functions in plant growth using cucumber seedling cuttings. Surface-sterilized cucumber seeds were grown in a plastic pot containing steamed vermiculite. After 7 days of cultivation, the upper part 2 cm in length of cucumber seedling, was cut and used as cucumber cuttings. The base of cutting stem was then dipped in a microcentrifuge tube containing 1.5ml of a bacterial suspension and incubated at $25^{\circ}C$ with a fluorescent light for 10 days. Number and length of developed adventitious roots from cucumber cuttings were examined. The seedling cuttings showed various responses to the isolates tested. Some isolates resulted in withering at the day of examination or in reduced number of roots developed. Several isolates stimulated initial development of adventitious roots showing more adventitious root hair number than that of untreated cuttings, while some isolate had more adventitious root hair number and longer adventitious roots than that of untreated control. Similar results were obtained from the trial with rose cuttings. Our results suggest that this bioassay method may provide a useful way for differentiating PGPR's functions involved in the development of root system.

Auxin Affects on Production of Adventitious Roots and Secondary Metabolites in Echinacea angustifolia (오옥신 처리가 에키네시아의 부정근 및 생리활성물질 생산에 미치는 영향)

  • Jang, Young Se;Cui, Hai Yan;Lee, Eun Jung;Kim, Hae Won;Paek, Kee-Yoeup
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.479-486
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    • 2012
  • The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L ${\alpha}$-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.

Production of Adventitious Ginseng Roots Using Biorectors

  • Yu, Kee-Won;Hahn, Eun-Joo;Paek, Kee-Yoeup
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.309-315
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    • 2000
  • Panax ginseng is an important medicinal plant that has been used worldwide for geriatric, tonic, stomachic, and aphrodisiac treatments. Ginsenosides contained in the ginseng root are the main substances having active functions for human body. The price of ginseng is very expensive due to a complex process of cultivation, and the yield of ginseng is limited, which cannot meet the demand of the increasing market. Researchers have applied plant biotechnology to solve the problems but there are still things to be determined towards ginsenoside production by large-scale adventitious root culture. In this experiment, 5 to 20 liter bioreactors were employed to determine optimal conditions for adventitious root culture and ginsenoside production of Panax gineng. Callus was induced from the ginseng root on MS agar medium containing 1.0 mg. $L^{-1}$ 2,4-D and 0.1 mg. $L^{-1}$ kinetin. Then the callus was cultured on MS agar medium supplemented with 2.0 mg. $L^{-1}$ IBA, 0.1 mg. $L^{-1}$ kinetin, and 30 g. $L^{-1}$ to induce adventitious roots. The maximum root growth and ginsenoside production were obtained in 1/2 MS medium. 2.0 mg. $L^{-1}$ naphthalene acetic acid resulted in greater root growth than 2.0 mg $L^{-1}$ indole-3-butyric acid. Ginsenoside content increased with 2.0 mg. $L^{-1}$ benzyl adenin or kinetin. High concentrations of benzyl adenin (above 3.0 mg. $L^{-1}$ ) decreased the adventitious root growth and ginsenoside productivity. N $H_{4}$$^{+}$ inhibited the ginsenoside accumulation, while high concentrations of $K^{+}$, $Mg_{2}$$^{+}$, and $Ca_{2}$$^{+}$ increased it. N $H_{4}$$^{+}$ at 0.5 and 1.0 times of the normal amount in 3/4 SH medium resulted in the greatest biomass increase, but the highest ginsenoside productivity was obtained when N $O_{3}$$^{-}$ was used as the sole nitrogen source in the medium. Most microelements at high concentrations in the medium inhibited the root growth, but high concentrations of MnS $O_4$enhanced the root growth. Root dry weight increased with increasing sucrose concentrations up to 50 g. $L^{-1}$ , but decreased from 70 g $L^{-1}$ Ginsenoside productivity was maximized at the range of 20 to 30 g. $L^{-1}$ sucrose. In the experiment on bioreactor types, cone and balloon types were determined to be favorable for both adventitious root growth and ginsenoside production. Jasmonic acid was effective for increasing ginsenoside contents and Rb group ginsenosides mainly increased. These results could be employed in commercial scale bioreactor cultures of Panax ginseng.x ginseng.

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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.

Adventitious Root Development and Ginsenoside Production in Panax ginseng, Panax quinquefolium and Panax japonicum

  • Han, Jung-Yeon;Kwon, Yong-Soo;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.147-152
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    • 2006
  • This work was carried out to establish adventitious root culture system in three Panax species (wild-grown P. ginseng, P. quinquefolium, and P. japonicum) to analyze their ginsenoside productivity. Adventitious roots were induced directly from segments of seedlings after cultured on MS(Murashige andSkoog 1962) solid medium containing 3.0 mg/l IBA. Omission of $NH_4NO_3$ from the medium greatly enhanced both the frequency of adventitious root formation and number of roots per explants in all the three Panax species. However, elongation of post-induced adventitious roots was enhanced on medium with $NH_4NO_3$. Two-step culture protocol: $NH_4NO_3$-free medium for first two weeks of culture, followed by $NH_4NO_3$ containing medium for further 4 weeks, greatly enhanced the fresh weight increase of adventitious roots in all the three ginseng species. The fresh weight of adventitious roots was high in P. quinquefolium and low in P. ginseng, followed by P. japonioum regardless of the composition of medium. Pattern and content of ginsenosides in adventitious roots differed among the three Panax species. Total ginsenoside content of adventitious roots in P. quinquefolium, P. ginseng, and p. japonicum was 8.03, 15.7 and 1.2 mg/g dry weight, respectively. Among the three speices, adventitious roots in P. quinquefolium produced hig-hamount of ginsenosides. The pattern of ginsenoside fractions between P. ginseng and P. quinquefolium was similar but the amount of ginsenoside differed between the two, While, in P japonicum, total ginsenoside content was very low and some ginsenosides such as ginsenoside Rb2 and Rf were not detected. Conclusively, we demonstrate that same culture condition was required for induction and elongation of adventitious roots of three ginseng species but growth of adventitious roots and their ginsenoside production were different among them.