Liquid Culture Systems for in vitro Plant Propagation

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Chapter 40 Cultivation of root cultures of Panax ginseng in different bioreactors and in temporary immersion - Comparison of growth and saponin production Tomáš Van�k, Lenka Langhansová* & Petr Maršík Institute of Organic Chemistry and Biochemistry, AS CR, Flemingovo nám. 2, 166 10, Praha 6, Czech Republic. *Author for correspondence; Fax: +420-220183574; E-mail: lenka@uochb.cas.cz Abstract: Different systems of large-scale cultivation of multiple adventitious roots of Panax ginseng C. A. Meyer were compared to cultivation in Erlenmeyer flasks. Adventitious roots were isolated from plantlets regenerated from somatic embryos and cultivated separately in liquid media. Multiplication of adventitious roots was performed in liquid Schenk and Hildebrandt (1972) medium containing 3 % sucrose, and 24.6 µmol indole-3-butyric acid. The highest saponin content of 28.51 mg g -1 of dry weight was found in adventitious roots cultivated in the RITA ® temporary immersion system (TIS). The best production of biomass was achieved in RITA ® vessels and standard Erlenmeyer flasks placed on rotary shaker, followed by the Applikon 3-litre bioreactor and a simple airlift reactor. Saponin production in Erlenmeyer flasks was 10.07 mg g -1 of the dry weight while the production in the Applikon 3-litre bioreactor was only 3.60 mg g -1 . Other bioreactor systems tested showed neither significant saponin production nor high biomass production. Key words: bioreactor, saponin, Panax ginseng, adventitious roots Abbreviations: IBA – indole-3-butyric acid; MS – Murashige and Skoog (1962); NAA – naphthalene acetic acid; SH – Schenk and Hildebrandt (1972); TIS – temporary immersion system 1. Introduction Panax ginseng C. A. Meyer (Araliaceae) is an herbaceous plant, which in Oriental medicine has a strong reputation since ancient times for being a tonic, regenerating, and rejuvenating, even though its pharmacological activity has not been fully elucidated. Ginseng grows wild in mountain areas, from Nepal to Manchuria, and from eastern Siberia to Korea but has been 539 A.K. Hvoslef-Eide and W. Preil (eds.), Liquid Culture Systems for in vitro Plant Propagation, 539–546. © 2005 Springer. Printed in the Netherlands.
540 Tomáš Vanek et al. overexploited. Recently it was reported that ginsenosides and polyacetylenes isolated from ginseng roots have cytotoxic activity (Newal et al., 1996). The current supply of ginseng mainly depends on field cultivation, which is a long and laborious process. Native ginseng plants need 5 – 7 years growth prior to harvest and the content of ginsenosides is low. In vitro mass production in large-scale systems seems to be potentially a more efficient alternative for the production of the bioactive components of ginseng. In our previous study we compared saponin production in different ginseng tissue cultures namely callus, suspension culture and adventitious roots. We found a full range of ginsenosides distributed analogous as in the roots of native plants in adventitious roots cultivated in liquid Schenk and Hildebrandt medium supplemented with 24.6 µmol IBA while the saponin content in callus as well as in cell suspension cultures was limited to major ginsenosides, Rb1 or Rg1 (Langhansová et al., 2002; Langhansova et al., 2003b). The aim of this study was to establish in vitro cultures producing ginsenosides from adventitious roots. We compared different cultivation systems to find a way for more efficient large-scale production. 2. Material and methods 2.1 Adventitious root induction Adventitious roots of Panax ginseng were isolated from plantlets regenerated from somatic embryos and rooted on 1/3-strength MS medium (Langhansova et al., 2003a). Separated roots were transferred to liquid MS medium supplemented with 1 µmol NAA and cultivation was carried out in 500 ml Erlenmeyer flasks on a rotary shaker (125 rpm). Root proliferation occurred on liquid SH medium supplemented with 24.6 µmol IBA (Choi et al., 2000). 2.2 Bioreactor cultures Adventitious roots cultures were cultivated in different bioreactors and temporary immersion systems (TIS) and in standard cultivation system in Erlenmeyer flasks:
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LIQUID CULTURE SYSTEMS FOR IN VITRO
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A C.I.P. Catalogue record for this
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Contents Contributing Authors xiii
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Liquid culture systems for in vitro
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Contributing Authors Adelberg, J. 4
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Preface Plant cell, tissue and orga
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Chapter 1 General introduction: a p
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General introduction 3 2. Cell susp
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General introduction 5 rooted in gr
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General introduction 7 (Winkelmann
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General introduction 9 the bioreact
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General introduction 11 (TIS), resu
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General introduction 13 Barz W, Rei
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General introduction 15 Hohe A, Win
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General introduction 17 Shimazu T &
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I. Bioreactors
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22 Wayne R. Curtis 1. Introduction
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24 Wayne R. Curtis headspace double
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26 Wayne R. Curtis that is required
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28 Wayne R. Curtis C L y � P O2
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30 Wayne R. Curtis since there is a
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32 Wayne R. Curtis 5 cm 30 cm 30 o
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34 Wayne R. Curtis Pˆ N � � me
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36 Wayne R. Curtis Radial Flow Impe
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38 Wayne R. Curtis CS = Medium diss
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40 Wayne R. Curtis Murashige T & Sk
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42 Anne Kathrine Hvoslef-Eide et al
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Chapter 4 Practical aspects of bior
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Practical aspects of bioreactor app
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Chapter 5 Simple bioreactors for ma
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Simple bioreactors for mass propaga
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96 K. Y. Paek et al. opportunity fo
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98 K. Y. Paek et al. disadvantages
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100 K. Y. Paek et al. 3.1 Dissolved
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102 K. Y. Paek et al. optimizing bi
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104 K. Y. Paek et al. cosmetics, wh
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106 K. Y. Paek et al. protocol, mor
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108 K. Y. Paek et al. use. In order
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110 K. Y. Paek et al. a b c d e f F
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112 K. Y. Paek et al. a b c d e f F
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114 K. Y. Paek et al. Escalona M, L
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116 K. Y. Paek et al. Son SH & Paek
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118 Seppo Sorvari et al. economical
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120 Seppo Sorvari et al. membrane w
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122 Seppo Sorvari et al. 3. Results
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124 Seppo Sorvari et al. D.O. 160 1
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Chapter 8 Cost-effective mass cloni
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Cost-effective mass cloning of plan
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144 Eun-Joo Hahn & Kee-Yoeup Paek 1
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146 Eun-Joo Hahn & Kee-Yoeup Paek F
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148 Eun-Joo Hahn & Kee-Yoeup Paek T
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150 Eun-Joo Hahn & Kee-Yoeup Paek 3
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152 Eun-Joo Hahn & Kee-Yoeup Paek E
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Chapter 10 Control of growth and di
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Control of growth and differentiati
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Chapter 11 Temporary immersion syst
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Temporary immersion system: a new c
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198 Elio Jiménez González 1. Intr
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200 Elio Jiménez González Several
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202 Elio Jiménez González Figure
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204 Elio Jiménez González exploit
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206 Elio Jiménez González weeks i
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208 Elio Jiménez González germina
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210 Elio Jiménez González Escalan
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Chapter 13 Use of growth retardants
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Use of growth retardants for banana
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Chapter 14 Somatic embryo germinati
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Somatic embryo germination of Psidi
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Somatic embryo germination of Psidi
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232 Tino Hempfling & Walter Preil N
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234 Tino Hempfling & Walter Preil 3
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236 Tino Hempfling & Walter Preil F
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238 Tino Hempfling & Walter Preil F
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240 Tino Hempfling & Walter Preil 4
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242 Tino Hempfling & Walter Preil R
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244 C. Damiano et al. Over the last
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246 C. Damiano et al. 2.2 Temporary
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248 C. Damiano et al. Table 3: Wate
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250 C. Damiano et al. hyperhydricit
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Chapter 17 Optimisation of growing
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Optimisation of growing conditions
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264 Katerina Grigoriadou et al. cul
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266 Katerina Grigoriadou et al. 2.4
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268 Katerina Grigoriadou et al. mic
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270 Katerina Grigoriadou et al. of
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272 Katerina Grigoriadou et al. 4.
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274 Katerina Grigoriadou et al. Tei
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276 Ch. Wawrosch et al. possesses s
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278 Ch. Wawrosch et al. 3. Results
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280 Ch. Wawrosch et al. References
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Chapter 20 Propagation of Norway sp
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Propagation of Norway Spruce 285 2.
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Propagation of Norway Spruce 287 su
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Propagation of Norway Spruce 289 5.
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Propagation of Norway Spruce 291 bl
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Propagation of Norway Spruce 293 H
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296 M. Vágner et al. 1995). In liq
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298 M. Vágner et al. maturation pe
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300 M. Vágner et al. mature somati
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302 M. Vágner et al. Acknowledgeme
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304 Christopher Hunter & Lionel Lev
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314 Michel Petitprez et al. 1. Intr
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316 Michel Petitprez et al. 2.2 QTL
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318 Michel Petitprez et al. Figure
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320 Michel Petitprez et al. Table 1
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322 Michel Petitprez et al. Gentzbi
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324 F. Afreen et al. 1. Introductio
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326 F. Afreen et al. precotyledonar
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328 F. Afreen et al. Dry mass (mg/e
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330 F. Afreen et al. under photoaut
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332 F. Afreen et al. Figure 3: a) C
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334 F. Afreen et al. the plant qual
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Chapter 25 Potential of flow cytome
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Potential of flow cytometry 339 dis
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Potential of flow cytometry 341 Fig
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Potential of flow cytometry 343 tha
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Chapter 26 Somatic embryogenesis of
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Somatic embryogenesis of Gentiana 3
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Chapter 27 Induction and growth of
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Induction and growth of tulip 361 a
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Induction and growth of tulip 363 T
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Chapter 28 Micropropagation of Ixia
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Micropropagation of Ixia 367 for PA
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Micropropagation of Ixia 369 Biomas
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Micropropagation of Ixia 371 Figure
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Chapter 29 Micropropagation of Rosa
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Micropropagation of Rosa 375 The BA
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Micropropagation of Rosa 377 Platfo
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Micropropagation of Rosa 379 3. Res
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Micropropagation of Rosa 381 The ro
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Micropropagation of Rosa 383 4. Dis
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Micropropagation of Rosa 385 Murash
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Chapter 30 Mass propagation of coni
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Mass propagation of conifer trees 3
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Chapter 31 Potentials for cost redu
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Potentials for cost reduction 405 c
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Potentials for cost reduction 409 s
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Chapter 32 A new approach for autom
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A new approach for automation 417 (
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A new approach for automation 419 p
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426 B. McAlister et al. 1. Introduc
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428 B. McAlister et al. 2.2 Multipl
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430 B. McAlister et al. Table 2: Mu
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432 B. McAlister et al. rest time -
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434 B. McAlister et al. Figure 3: M
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436 B. McAlister et al. Average mul
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438 B. McAlister et al. Table 4: Co
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440 B. McAlister et al. Semi-solid
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442 B. McAlister et al. Escalona M,
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444 Jeffrey Adelberg including Host
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446 Jeffrey Adelberg BioSystems (Ho
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448 Jeffrey Adelberg more prolific
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450 Jeffrey Adelberg vessel of liqu
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452 Jeffrey Adelberg Figure 4: Labo
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454 Jeffrey Adelberg constituted a
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456 Jeffrey Adelberg Acknowledgemen
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Chapter 35 Aeration stress in plant
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Aeration stress in plant tissue cul
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476 Hans R. Gislerød et al. 1. Int
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478 Hans R. Gislerød et al. biorea
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480 Hans R. Gislerød et al. A low
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482 Hans R. Gislerød et al. Table
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484 Hans R. Gislerød et al. common
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Page 534 and 535: Cultivation of root cultures of Pan
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Page 542 and 543: Optimisation of Panax ginseng liqui
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Page 560 and 561: Production of taxanes 569 2. Materi
Page 562 and 563: Production of taxanes 571 Figure 2:
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Page 566 and 567: Acknowledgments The editors thank t
Page 568 and 569: 578 Contents 130, 131, 133, 134, 13
Page 570 and 571: 580 Contents poplar, see Populus Po
Page 572 and 573: 582 Contents bubble aerated 165 bub
Page 574 and 575: 584 Contents ginsenoside content, p
Page 576 and 577: 586 Contents plant growth regulator
Page 578: 588 Contents -free microcorms 71 -f
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