Liquid Culture Systems for in vitro Plant Propagation

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30 Wayne R. Curtis since there is a drop in oxygen to the surface of the tissue due to external mass transfer, and a reduction in oxygen within the tissue due to consumption and limitations in diffusional mass transfer. Oxygen supplementation is not a panacea for increasing O2 availability. Oxygen supplementation can be quite costly – particularly for the long culture times required for plant tissue culture. Advances in pressure swing adsorption (PSA) now provide a relatively inexpensive means of oxygen supplementation to a process of the scale under discussion for commercial micropropagation. Another processing approach for oxygen supplementation worth noting is the separate sparging of high-purity oxygen in very small bubbles. The advantage of this approach is that the driving force for transport from these bubbles will be five times greater than ambient air, and much higher than would be accomplished if the oxygen was mixed with air sparging. The production of small bubbles is useful since the small radius of curvature will create bubbles with high internal pressure that are not prone to coalescence with the sparged air bubbles. Therefore, separate sparging and small gas bubbles avoid dilution of the driving force for oxygen transfer that results when oxygen is mixed with air. The relatively small scale of micropropagation (relative to industrial fermentation) tends to reduce the impact of costs for oxygen supplementation. Table 2: Influence of media components on media dissolved oxygen 2A – Bunsen Coefficient contributions (Schumpe et al., 1981) Ki / Hi (l mol -1 sucrose ) 44 glucose 8.8 + NH4 6.3 - NO3 6.2 K + 5.0 - H2PO4 4.5 2B – Calculated equilibrium dissolved oxygen in plant culture media (Eqn. 8) O 2 @ 25 o C (mg l -1 ) Water 8.24 MS (30 g sucrose l -1 ) 7.92 DCR (20 g sucrose l -1 ) 8.04 B5 (20 g sucrose l -1 ) 8.01 MS-Murashige and Skoog, 1962; DCR-Gupta and Durzan, 1985; B5-Gamborg et al., 1968.
Application of Bioreactor Design Principles 31 5. Circulation and micropropagule suspension The relatively rapid sedimentation velocity of plant tissues results in a need for adequate circulation to provide suspension. The sedimentation rate of a nearly spherical object can be estimated from Stokes law, media 2 p ( �tissue � �media) g � d vsediment � (Eqn. 7) 18� � however, the sedimentation rate is often sufficiently fast that the laminar flow assumptions for this equation are no longer valid and a more involved calculation using a (laminar-turbulent) drag coefficient is needed (Singh and Curtis, 1994). This equation, none the less, provides the important functional dependencies. The density of the media (�media) is roughly the density of water, therefore, the more dense the tissues, the faster they will sediment. We observed the sedimentation rate of oak embryos to correspond to a tissue density of about 1.05 g ml -1 , which was considerably faster than unorganized plant cell aggregates (Singh and Curtis, 1994). Gravity (g) is a constant, and the effective viscosity of the media (�media) will increase when there are more cells present in the suspension, or if the suspended plant tissues are elongated (Curtis and Emery, 1993). The diameter of the suspending particle (dp) is important for this analysis because it has a square dependence – and because plant micropropagules such as somatic embryos are extremely large by comparison to ‘typical’ microbial suspensions. For example, a suspension of embryos could be expected to sediment a 1 cm in a non-agitated culture flask in about 1 second. By comparison, a yeast suspension would require about 1 hour to sediment over the same depth. Preventing sedimentation of plant tissues to the bottom of a bioreactor can become a problem due to their rapid sedimentation rate and the relatively low energy inputs typically used for growth of plant tissues in bioreactors. To study the issue of somatic embryo suspension more thoroughly, two ~ 30-l vessels were constructed. Polystyrene beads (�=1.05 g ml -1 ) with a diameter of 3.2 mm (vsediment = 5.1 cm s -1 ) were used to represent somatic embryos in these suspension studies. The first ‘bioreactor’ used only gas sparging and had a geometrical design based on a recent low-cost bioreactor patent (Curtis, 2001) and is shown schematically in figure 3.
Page 2 and 3: LIQUID CULTURE SYSTEMS FOR IN VITRO
Page 4 and 5: A C.I.P. Catalogue record for this
Page 6 and 7: Contents Contributing Authors xiii
Page 8 and 9: Liquid culture systems for in vitro
Page 12 and 13: Contributing Authors Adelberg, J. 4
Page 16 and 17: Preface Plant cell, tissue and orga
Page 18 and 19: Chapter 1 General introduction: a p
Page 20 and 21: General introduction 3 2. Cell susp
Page 22 and 23: General introduction 5 rooted in gr
Page 24 and 25: General introduction 7 (Winkelmann
Page 26 and 27: General introduction 9 the bioreact
Page 28 and 29: General introduction 11 (TIS), resu
Page 30 and 31: General introduction 13 Barz W, Rei
Page 32 and 33: General introduction 15 Hohe A, Win
Page 34 and 35: General introduction 17 Shimazu T &
Page 36 and 37: I. Bioreactors
Page 38 and 39: 22 Wayne R. Curtis 1. Introduction
Page 40 and 41: 24 Wayne R. Curtis headspace double
Page 42 and 43: 26 Wayne R. Curtis that is required
Page 44 and 45: 28 Wayne R. Curtis C L y � P O2
Page 48 and 49: 32 Wayne R. Curtis 5 cm 30 cm 30 o
Page 50 and 51: 34 Wayne R. Curtis Pˆ N � � me
Page 52 and 53: 36 Wayne R. Curtis Radial Flow Impe
Page 54 and 55: 38 Wayne R. Curtis CS = Medium diss
Page 56 and 57: 40 Wayne R. Curtis Murashige T & Sk
Page 58 and 59: 42 Anne Kathrine Hvoslef-Eide et al
Page 76 and 77: Chapter 4 Practical aspects of bior
Page 78 and 79: Practical aspects of bioreactor app
Page 94 and 95: 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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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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