Liquid Culture Systems for in vitro Plant Propagation

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528 Dirk Wilken et al. running at 60 rpm. The culture temperature was 26 °C. Both bioreactors were inoculated with 20 g l -1 cell fresh weight and harvested after 19 days. 2.3 Temporary immersion system (TIS) A temporary immersion system was constructed using 5-litre glass twin vessels. Under standard conditions the system was run with 2 litres of medium that was changed every 2 or 3 weeks. The TIS vessels were inoculated with 30 g of in vitro-grown shoots. Shoots were immersed every 4 h for 5 min. The temperature and the light intensity were as given above. The cultures were harvested after 8 weeks. 2.4 Field grown plants Fabiana imbricata was collected in the Western Andean slopes near Altos de Chillan, Las Trancas, Chile. Lavandula officinalis and Hypericum perforatum plants were obtained from the Botanical Garden of the University of Leipzig. For analysis of the leaf oil composition of Lavandula officinalis commercially dealed material originated from Corsica. Cymbopogon citratus was collected in Bayamo, Cuba. 2.5 Analysis of secondary plant metabolites Contents of bioactive compounds were determined in powdered tissue samples that were oven-dried at 40 °C. The metabolites of interest were: rosmarinic acid and essential oil in Lavandula, hypericin in Hypericum, �and �-citral in Cymbopogon and oleanolic acid in Fabiana. 2.5.1 Sample preparation and determination of rosmarinic acid According De-Eknamkul (1984), Gracza und Ruff (1984) and Lopez et al. (1994) a method for determination of the content of rosmarinic acid in cultures of Lavandula was established. Samples of 20 mg of dried and powdered plant material were extracted two times with 3 ml methanol in an ultrasonic bath at room temperature for 10 minutes. The filtrates were collected, filtrated and analysed by HPLC. A Gynkotek HPLC system (Dionex) equipped with a diode array detector (UVD 340S) and a LiChrocart RP18 Supersphere Colomn (250mm x 4 mm; 4 µm) was used. A mixture of methanol/water/phosphoric acid (50/59.7/0.3; v/v) was used as eluant at a flow rate of 0.5 ml min -1 . All measurements were carried out at 30°C under isocratic conditions. Rosmarinic acid was detected at 333 nm and showed a
Comparison of secondary plant metabolite production 529 retention time of 9.8 minutes. A solution of 1 mg ml -1 rosmarinic acid was used as standard. 2.5.2 Determination of the composition of essential oil from Lavandula leaves To obtain the essential oil an apparatus described in the European Pharmacopeia was used (European Pharmacopeia 2002). 20 g of dried in vitro plants or dried leaves from in vivo or field plants were heated with 500 ml water and steam distilled for 3 hours. Mass spectra of the essential oil obtained were recorded on a Finnigan MAT ITD-800 gas chromatography – mass spectroscopy instrument (injection volume: 0.2 µl, Split 1:100). The GC column used was DB1 60 m x 0.25 mm ID x 0.25 µm film thickness. It was heated from 60°C to 220°C with a speed of 2°C min -1 . Helium flow-rate was 1 ml min -1 . The ionization mode used was electron impact ionization with an ion trap temperature of 200°C. 2.5.3 Sample preparation and determination of hypericin The content of hypericin in the plant biomass was analysed using a modification of a method developed by Kartnig et al. (1996). Samples of 100 - 400 mg of dried and powdered plant material were extracted three times with 3 ml methanol in an ultrasonic bath at room temperature. The filtrates were collected, filtered and analysed by HPLC. A Gynkotek HPLC system (Dionex) equipped with a diode array detector (UVD 340S) and a LiChrocart RP18 Supersphere Colomn (250 mm x 4 mm; 4 µm) were used. A mixture of methanol/ethylacetate/phosphate (3.6/1/1.2; v/v) was used as eluant with a flow of 0.9 ml min -1 . All measurements were carried out at 30°C under isocratic conditions. Hypericin was detected at 595 nm. A solution of 0.53 mg hypericin in 10 ml methanol was used as standard. Under the conditions described a retention time of 18 min was observed. 2.5.4 Sample preparation and determination of �- and �-citral Samples of 10-20 mg dried and powdered plant material were extracted each with 1.0 ml methanol 10 min in an ultrasonic bath. The extracts were filtered (0.4 µm) and analysed by HPLC. A Nucleosil 100-5 column (C18, 250 x 3 mm) produced by Knauer, Berlin was used. The mobile phase consisted of methanol and water (70/30; v/v) with a flow rate of 0.8 ml min -1 . The column was heated to 50 °C. All measurements were carried out at isocratic conditions. Citral was detected at 240 nm. Under the conditions
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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 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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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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Page 472 and 473: 476 Hans R. Gislerød et al. 1. Int
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Page 520 and 521: 526 Dirk Wilken et al. 1. Introduct
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Page 526 and 527: 532 Dirk Wilken et al. packed cell
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Page 532 and 533: Chapter 40 Cultivation of root cult
Page 534 and 535: Cultivation of root cultures of Pan
Page 540 and 541: Chapter 41 Optimisation of Panax gi
Page 542 and 543: Optimisation of Panax ginseng liqui
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Page 558 and 559: Chapter 43 Production of taxanes in
Page 560 and 561: Production of taxanes 569 2. Materi
Page 562 and 563: Production of taxanes 571 Figure 2:
Page 564 and 565: Production of taxanes 573 various T
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
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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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