Liquid Culture Systems for in vitro Plant Propagation

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232 Tino Hempfling & Walter Preil Netherlands, exceeded 64 million Euro in 2002. The increase was 30.9 % compared to 2001 (Anonymous, 2003). In vitro culture procedures including 22 media formulations for Phalaenopsis were listed by Arditti and Ernst (1993), presenting an overview on the state of knowledge at that time. The application of standard protocols for micropropagation on solid medium on average results in a three-fold shoot multiplication rate after an eight-week subculture period. More recently, liquid systems including bioreactors or temporary immersion techniques were successfully applied giving rise to higher multiplication rates (Park et al., 1996, 2000; Samson et al., 2000). Most of agar gelled media for Phalaenopsis micropropagation are supplemented with BAP as cytokinin source. A few publications stress the positive effects of TDZ on protocorm or adventitious bud regeneration (Ernst, 1994; Chen and Piluek, 1995; Chen et al., 2000). As far as we know, no experiments have been published on the effects of TDZ in liquid cultures of Phalaenopsis. The present study was carried out to investigate adventitious shoot multiplication and rooting in a temporary immersion system (TIS) using five-litre twin glass vessels. The effects of medium substitution after two or four weeks were specifically determined. Further, the influence of TDZexposure for 7 or 12 weeks on shoot size, multiplication rate and fresh weight was investigated. During the rooting phase various immersion frequencies were applied to determine optimum conditions. 2. Material and methods 2.1 Multiplication In vitro grown shoots of Phalaenopsis cv. Jaunina (Breeder: Wolfgang Bock, Bremen, Germany) were used as inoculum. Per five-litre glass vessel 70–72 shoots (1 – 3 cm length; total fresh weight: 20g) were incubated. Each vessel was connected through a silicone tube with another vessel (Figure 1 A) containing 2 litres of liquid MS-medium of ½ strength macro- and micro-elements (Murashige and Skoog, 1962), supplemented with 170 mg l -1 NaH2PO4, 100 mg l -1 myo-inositol, 0.5 mg l -1 nicotinic acid, 0.5 mg l -1 pyridoxine, 0.1 mg l -1 thiamine HCl, 2.0 mg l -1 glycine, 1000 mg l -1 peptone and 20 g l -1 sucrose, as published by Chen et al. (2000). Additionally 0.5 mg l -1 thidiazuron (TDZ) was added. The liquid medium was drained from the ‘medium-vessel’ into the ‘plantvessel’ and vice versa by means of compressed air being passed through a three-way solenoid valve controlled by a programmable timer as described
Application of temporary immersion in Phalaenopsis 233 by Escalona et al. (1999). The immersion of shoots was adjusted to eight times per day for ten minutes each. Spent medium blackened by phenolic compounds (Figure 1 A) was substituted in intervals of two or four weeks. The experiments using the twin vessels were repeated three times each. The multiplication rate and increase of fresh weight were determined twelve weeks after inoculation. An overview of the experimental design is given in figure 1 B. Control cultures were grown on agar gelled medium (Tanaka and Sakanishi, 1985) supplemented with 10 mg l -1 BAP and 10 % (v/v) homogenised bananas. 2.2 Rooting In the first series of experiments shoots of 4 – 7 cm size from TIS cultures were exposed to TDZ-free medium supplemented with 0.5 and 1.0 mg l -1 IAA or NAA. In the main experiments 1.0 mg l -1 IAA-containing medium was used. Each vessel was inoculated with 72 – 74 shoots of 70 g total fresh weight. The immersion frequency was adjusted to two, four or six times per day for ten minutes periods. The experiments using the twin vessels were repeated three times each. The percentage of rooted plants and the increase in fresh weight were determined after eight weeks. Figure 1: (A) Twin glass vessels (5 l) inoculated with 20 g shoot fresh weight of Phalaenopsis cv. Jaunina. After two weeks the medium was blackened by phenolic compounds. (B) Experimental design of a series of twin glass vessels after 12-week culture.
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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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Page 188 and 189: Temporary immersion system: a new c
Page 206 and 207: 198 Elio Jiménez González 1. Intr
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Page 214 and 215: 206 Elio Jiménez González weeks i
Page 216 and 217: 208 Elio Jiménez González germina
Page 218 and 219: 210 Elio Jiménez González Escalan
Page 220 and 221: Chapter 13 Use of growth retardants
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Page 268 and 269: 264 Katerina Grigoriadou et al. cul
Page 270 and 271: 266 Katerina Grigoriadou et al. 2.4
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Page 282 and 283: 278 Ch. Wawrosch et al. 3. Results
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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 407 c
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Potentials for cost reduction 409 s
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Potentials for cost reduction 411 N
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Potentials for cost reduction 413 G
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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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A new approach for automation 421 T
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A new approach for automation 423 D
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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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486 Hans R. Gislerød et al. can be
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488 Hans R. Gislerød et al. 4.1 Li
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490 Hans R. Gislerød et al. 4.3 Ca
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492 Hans R. Gislerød et al. Morten
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494 Han Bouman & Annemiek Tiekstra
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V. Biomass for Secondary Metabolite
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510 E. Gontier et al. 1. Introducti
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512 E. Gontier et al. 2. Materials
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514 E. Gontier et al. developed dra
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516 E. Gontier et al. 3.3 Establish
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518 E. Gontier et al. Fresh weight
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520 E. Gontier et al. 3.5 Effect of
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522 E. Gontier et al. 3.6 Scale up
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524 E. Gontier et al. Lorenzo JC, G
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526 Dirk Wilken et al. 1. Introduct
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528 Dirk Wilken et al. running at 6
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530 Dirk Wilken et al. described a
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532 Dirk Wilken et al. packed cell
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534 Dirk Wilken et al. bioactive co
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536 Dirk Wilken et al. Fowler MW (1
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Chapter 40 Cultivation of root cult
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Cultivation of root cultures of Pan
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Chapter 41 Optimisation of Panax gi
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Optimisation of Panax ginseng liqui
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558 S. M. Nalawade et al. performan
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560 S. M. Nalawade et al. Plantlets
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562 S. M. Nalawade et al. leaves on
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564 S. M. Nalawade et al. roots (Fi
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Chapter 43 Production of taxanes in
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Production of taxanes 569 2. Materi
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Production of taxanes 571 Figure 2:
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Production of taxanes 573 various T
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Acknowledgments The editors thank t
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578 Contents 130, 131, 133, 134, 13
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580 Contents poplar, see Populus Po
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582 Contents bubble aerated 165 bub
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584 Contents ginsenoside content, p
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586 Contents plant growth regulator
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588 Contents -free microcorms 71 -f
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