• Title/Summary/Keyword: Hairy root

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Selection of Optimal Biotic Elicitor on Tropane Alkaloid Production of Hairy Roots in Scopolia parviflora Nakai (미치광이풀 모상근 배양에서 Tropane Alkaloids 생산성 증진을 위한 최적 생물학적 엘리시터 선발)

  • Jung, Hee-Young;Kang, Seung-Mi;Kang, Young-Min;Kim, Yong-Duk;Yang, Jae-Kyung;Chung, Young-Gwan;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.358-363
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    • 2003
  • ScopoIamine and hyoscyamine which belong to tropane alkaloids are the pharmaceutically valuable anticholinergic drugs. In order to increase the productivities, the effects of elicitation were investigated during hairy root cultures of Scopolia. parviflora. Biotic elicitors originated from 3 fungi and 1 yeast were prepared as homogenate and supernatant and added to 3-week-old cultures. Both of homogenate and supernatant of Candida albicans elicitors increased the scopolamine production. The production of hyoscyamine was enhanced by homogenate of Fusarium solani and supernatant of C. albicans. Most of the other fungal elicitors were also improved on the tropane alkaloid production compared to non-treatment. Among the elicitors tested, C. albicans was proved the optimal biotic elicitor on tropane alkaloids production. These results will be served mass production of tropane alkaloids by large-scale production.

Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures. (Red Beet의 모상근 배양을 이용한 천연색소인 Betacyanin 생산의 최적화)

  • Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.435-441
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    • 1998
  • Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

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Production of secondary metabolites by tissue culture of Artemisia annua L. (Artemisia annua L.의 조직배양을 이용한 이차대사 산물의 생산)

  • Kim, Nam-Cheol;Kim, Jeong-Gu;Lim, Hyung-Joon;Hahn, Tae-Ryong;Kim, Soo-Un
    • Applied Biological Chemistry
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    • v.35 no.2
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    • pp.99-105
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    • 1992
  • Artemisia annua contains the antimalarial principle, artemisinin. The possibility of the production of this compound through tissue culture technique was studied. The optimum combinations of hormones for the induction of callus were p-chlorophenoxyacetic acid(pcPA) and 6-benzylaminopurine(BAP) or pcPA and N-isopentenylaminopurine(2iP) in 0.05 mg/l each. For the growth of callus, the same combination of pcPA and BAP was optimum in concentrations of $1.0\;{\mu}M\;and\;0.5\;{\mu}M$, respectively, and the optimal concentration of sucrose was also found to be 2%(w/v). Tissue culture from the crown gall grew faster than normal callus. In the suspension culture broth and the cells of normal callus or Agrobacterium-transformed tumors, arteannuic acid and 11,12-dihydroarteannuic acid were found together with common phytosterols, whereas artemisinin was not found.

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Effects of Organic Fertilizers and Green Manure Crops on Growth Responses and Yields of Astragalus membranaceus Bunge (황기의 녹비작물 작부체계 활용 유기재배 시 생장반응과 수량에 관한 연구)

  • Song, Beom-Heon;Lee, Kyung-A;Chang, Yoon-Kee;Kim, Young-Gook;Ahn, Tae-Jin;Ahn, Young-Seob;Park, Chung-Beom
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.2
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    • pp.83-89
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    • 2011
  • This studies were conducted to investigate the growth responses and yields of Astragalus membranaceus Bunge which were grown with organic cultivation using two organic fertilizers and two green manure crops of hairy vetch and rye in 2008 and 2009. The lengths of stems and roots were not clearly different between organic and conventional cultivations. The root diameters and the branch numbers were slightly thicker and more in all treatments of organic cultivation than that of conventional cultivation in 2008, although they were not significantly different between two cultivating methods in 2009. The dry weights of shoot and root were heavier on organic cultivation than those on conventional cultivation. The T/R ratios were commonly higher in organic culture, showing more clearly in 2008 than 2009. The yields were generally higher in organic cultivation than those in conventional cultivation in 2008 although the effect did not clear on yield in 2009. The organic cultivation using of organic fertilizers and green manure crops would be new method to have safety and qualitative products. The techniques of organic cultivation for Astragalus membranaceus Bunge would be studied more on utilizing the natural organic resources.

Production of Monoclonal Antibody about Specific Key Enzyme of Hyoscyamine $6{\beta}-Hydroxylase$ (H6H) in Scopolia parviflora

  • Kang, Young-Min;Jung, Hee-Young;Kang, Seung-Mi;Jin, Byung-Rae;Lee, Sang-Chul;Lee, Byung-Hyun;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.135-140
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    • 2004
  • Total RNAs were isolated from cultured roots of Scopolia parviflora, $poly(A)^+$ RNA was obtained through the mRNA purification, cDNA library of Hnh6h was constructed. Recombinant baculoviruses in Spodoptera frugiperda (Sf) cells were constructed by use of the transfer vector pBacPAK, which has the AcNPV sequence under the polyhedrin promoter. The expression vector carrying Hnh6h gene was transferred to S. parviflora and obtained transgenic hairy root lines. Our results confirmed the over expression of the H6H protein was used by anti-pBacPAK about cDNAs of S. parviflora. This study will served for production of tropane alkaloids by metabolic engineering.

Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Plant cell culture strategies for the production of natural products

  • Ochoa-Villarreal, Marisol;Howat, Susan;Hong, SunMi;Jang, Mi Ok;Jin, Young-Woo;Lee, Eun-Kyong;Loake, Gary J.
    • BMB Reports
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    • v.49 no.3
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    • pp.149-158
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    • 2016
  • Plants have evolved a vast chemical cornucopia to support their sessile lifestyles. Man has exploited this natural resource since Neolithic times and currently plant-derived chemicals are exploited for a myriad of applications. However, plant sources of most high-value natural products (NPs) are not domesticated and therefore their production cannot be undertaken on an agricultural scale. Further, these plant species are often slow growing, their populations limiting, the concentration of the target molecule highly variable and routinely present at extremely low concentrations. Plant cell and organ culture constitutes a sustainable, controllable and environmentally friendly tool for the industrial production of plant NPs. Further, advances in cell line selection, biotransformation, product secretion, cell permeabilisation, extraction and scale-up, among others, are driving increases in plant NP yields. However, there remain significant obstacles to the commercial synthesis of high-value chemicals from these sources. The relatively recent isolation, culturing and characterisation of cambial meristematic cells (CMCs), provides an emerging platform to circumvent many of these potential difficulties.

Phytochemical Studies on Astragalus Root(1) - Saponins

  • Kim, Ju-Sun;Yean, Min-Hye;Lee, Eun-Ju;Kang, Sam-Sik
    • Natural Product Sciences
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    • v.14 no.1
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    • pp.37-46
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    • 2008
  • From the 70% EtOH extract of the roots of Astragalus membranaceus (Leguminosae), fifteen saponins were isolated and identified as astragaloside I (1), isoastragaloside II (2), astragaloside II (3), agroastragaloside I (4), cyclogaleginoside B (5), cycloaraloside A (6), brachyoside B (7), agroastragaloside II (8), astragaloside III (9), astragaloside IV (10), astramembranoside A (11), astramembranoside B (12), cylocanthoside E (13), cyclounifolioside B (14) and azukisaponin V methyl ester (15) by spectroscopic methods. Ten compounds 1 - 3, 5 - 7, 9 - 11 and 14 have cycloastragenol as an aglycon, and four compounds 4, 8 , 12, and 13 have cyclocanthogenin as an aglycon. The hairy roots of A. membranaceus were shown to produce previously unreported cycloartane-type saponins such as agroastragalosides I (4) and II (8) and cycloastragenol $3-O-{\beta}-D-xyloside$ (5), together with the known saponins. This is the first report of these saponins (4, 5, and 8) from the intact plant. Although the occurrence of the oleanane-type triterpene saponin, azukisaponin V methyl ester (15), in Astragalus plants has been demonstrated by others, this is the first report of the azukisaponin V methyl ester (15) from the Astragalus plants.

First Report of Bacterial Root Rot Caused by Serratia plymuthica on Panax ginseng (Serratia plymuthica에 의한 인삼 세균뿌리썩음병 발생)

  • Jung, Won Kwon;Kim, Young Soo;Choi, Jin Kook;Kim, Seung-Han;Jang, Myeong-Hwan;Kwon, Tae Lyong;Jeon, Yongho
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.332-338
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    • 2018
  • In August of 2011, a wilting disease of ginseng was observed at Bongwha, Gyeongbuk province, Korea. Affected plants initially show withering symptoms on leaves of ginseng. As the disease progresses, withering leaves spread downward, eventually encompassing the whole plant. Leaves lose vigor but remain pale green. Symptoms of roots were brown, and soft rots characterized by moist and watery decay of the whole ginseng root, which initiated as small brown, water-soaked lesions of hairy roots and enlarged to the entire roots. The causal organism isolated from the infected roots was identified as Serratia plymuthica based on its physiological and biochemical characteristics, by cellular fatty acid composition (GC-FAME), the utilization of carbon sources (BioLog System), and 16S rRNA sequence of the isolated bacterium were 99% homologous to those of Serratia plymuthica strains. Artificial inoculation of the bacterium produced the same brown or soft rot symptoms on the ginseng roots, from which the same bacterium was isolated. This is the first report of bacterial root rot caused by the Serratia plymuthica in ginseng in Korea. Serratia plymuthica has been used as antagonistic microorganism for biological control on several crop plants. But it was proved pathogen of ginseng at humid condition in this study.