Liquid Culture Systems for in vitro Plant Propagation

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50 Anne Kathrine Hvoslef-Eide et al. 2.7 Microprocessors, computers and software Each bioreactor is controlled by a Personal Computer (PC). The main task of this computer is to be the user interface and for data logging. The user interface allows the operators to set parameters for the controller, specify set points, perform sensor calibration and set up other experimental properties. The set points can be specified to be constant during the experiment or to vary along a diurnal cycle, where the set points are freely specified for each of the 24 hours. Every actuator can also be manually controlled during the experiment to allow for the operators to interact actively with the process. Logged data are displayed as graphs. (Bakken, 1992; Kolstad, 1991; Myhre, 1991). However, the PC does not take an active part in the control of the actuators; this is under the control of a dedicated microprocessor. This processor is placed in an electronic rack together with sensor amplifiers, power supply, actuator drivers and the gas mixer (Unpublished electronics and software). We developed the electronics and programs for this project. The electronics hardware is housed in a rack, one for each bioreactor. The racks are self-contained units, which make the system independent of the ever-changing technology of the PC world. The control processor communicates over a RS232 serial line with the PC or any other RS232equipped system by a simple protocol developed for this project. This protocol and the interaction between the PC and the controller for parameter transfer; data logging etc. are described in more detail in Olsen and Heyerdahl (1994). Both the electronics and software of the control processor are substantially more simple than those of the PC and probably more stable and reliable than the PC. Since the critical online part of the system is external to the PC, the PC may malfunction without affecting the experiment. 3. Biological material and methods when starting an experiment When starting experiments in the bioreactors, it is important to perform the experiment with the same starting material to get homogenous and comparable suspension cultures. The whole concept of six identical bioreactors is to facilitate factorial experiments to reveal differences between treatments and possible interactions. In our experience, the same genotype may vary so much from batch to batch and during a year, that experiments need to be repeated in the bioreactors both in time and space to be reliable. Generally we follow the same procedure at all times: inoculate one
Bioreactor design for propagation 51 bioreactor and multiply the cells in culture, before inoculating all bioreactors with the same large-scale cell culture. 4. Results and Discussion 4.1 Comparing Cyclamen cell growth in Erlenmeyer flasks and in our bioreactor We have performed comparisons using identical Cyclamen persicum Mill cultures simultaneously, in both Erlenmeyer flasks and our bioreactors. We started the experiment with the same inoculum for both culture systems and using the same initial cell density and the same proliferation medium (Schwenkel and Winkelmann, 1998; Winkelmann et al., 1998), following the procedures described in Hvoslef-Eide and Munster (1998; 2001). The results during the first two-week growth period in four bioreactors showed significantly better growth in bioreactors; by fresh weight, dry weight, packed cell volume (PCV) and cluster viability measured by FDA staining (Widholm, 1972) of living cells (Figure 4). The standard deviation is less in the bioreactors than in flasks, indicating a more uniform and predictable culture in bioreactors in our system. Erlenmeyer flasks are probably more variable because of the inability to control the various important environmental parameters. Preil and co-workers (Preil et al., 1988; Preil, 1991) have shown the same improved growth pattern for poinsettia cell cultures using Braun Biostat bioreactors compared with Erlenmeyer flasks when the bioreactors were depleted of excess CO2 that may build up in a closed (recirculated gas) silicone tube system. Hohe et al. (1999a) found better growth in Erlenmeyer flasks of Cyclamen persicum cells compared to Applikon bioreactors (2 l) when they allowed the CO2 to build-up in the headspace. However, in later experiments, Hohe et al. (1999b; 2001) depleted CO2 from the circulating gas mixture and obtained improved growth in the bioreactors. This measure is not necessary in our bioreactors, since the aeration system is exhaustive and not in a closed circuit. We have investigated the gas permeability of the silicone tubes (Hvoslef-Eide and Munster, 1997) and found that the permeability could vary between tubes of different bioreactors, with build-up of CO2 and differences in growth of cells between identical bioreactors as a consequence. We concluded that the tubing age and number of autoclave cycles to which they have been subject, could affect their permeability and now always change the tubes in all six bioreactors at the same time after this.
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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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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 46 and 47: 30 Wayne R. Curtis since there is a
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
Page 96 and 97: Simple bioreactors for mass propaga
Page 110 and 111: 96 K. Y. Paek et al. opportunity fo
Page 112 and 113: 98 K. Y. Paek et al. disadvantages
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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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