Liquid Culture Systems for in vitro Plant Propagation

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28 Wayne R. Curtis C L y � P O2 � C* � (Eqn. 5) H It is important to note that equilibrium dissolved oxygen (C * ) is almost never measured; instead, probes are calibrated to give a percentage readout of this complex variable. Each of the parameters of oxygen mole fraction (yO 2 ), system pressure (P) and Henry’s Law coefficient (H) present different approaches to increasing oxygen availability. Bioreactor pressure will often enhance oxygen availability as an indirect consequence of bioreactor operation. In situ sterilisable bioreactors are pressure-rated autoclaves (sterilized @ ~18 p.s.i., 120 o C). The typical approach of operating at a head pressure of 5-10 p.s.i. not only reduces contamination risks from leaks, but can nearly double the equilibrium dissolved oxygen (1 atmosphere = 14.7 p.s.i.). In addition, the hydrostatic pressure (�H 2 0·g·depth) can be a significant contributor to oxygen transfer driving force (1 atmosphere � 33 feet of water). These observations have interesting implications for liquid culture micropropagation. The productivity of a reasonably efficient somatic embryogenic culture can generate large numbers of embryos in bioreactors smaller than 1000 l. Since scale-up would take place in tanks much smaller than traditional fermentation, the depth effect will be minimal. Use of pressurization (beyond autoclave pressures) is usually avoided due to the prohibitive cost of pressure vessels (Curtis, 1999). However, since ‘large-scale’ micropropagation can be implemented at a small scale, the pressure rating of the vessel (and associated steel costs) will not dominate overall equipment costs, and use of pressure could be economically feasible if it provides a substantial improvement in micropropagule development. It is also apparent from equation 5, that an increase in the Henry’s Law coefficient (H) will reduce the equilibrium dissolved oxygen pressure. Temperature has the greatest influence on H. As the temperature increases, H increases as thermal vibrations exclude oxygen, which decreases the maximum amount of oxygen that can dissolve in the media. Colder temperatures, therefore, favor oxygen solubility (15, 20, 25 and 30 o C correspond to 315, 283, 258, and 236 �mol O2 respectively in water). To take advantage of the increased oxygen solubility at lower temperatures, an organism must be able to sustain metabolism rates. This is not generally the case for terrestrial plant tissues, therefore, a reduction in temperature will usually significantly reduce the BOD. A suppression of tissue development rates is generally not desirable since it will increase the time-frame required to produce micro-propagules. This observation may, however, be useful if it is desirable to maintain aerobic respiration within a tissue by simultaneously
Application of Bioreactor Design Principles 29 reducing oxygen demand and increasing oxygen availability through growth at reduced temperatures. While the temperature dependence of oxygen solubility in water is widely known, the dependence of dissolved oxygen on media components receives less attention. Both the presence of inorganic salts and sugars has predictable effects on oxygen solubility. The approach to quantitatively assessing the impact of media components on solubility is more readily determined from an alternative to H that is referred to as the Bunsen coefficient. The Bunsen coefficient (�) is defined as the volume of oxygen dissolved per volume of media (calculated to a reference state of 0 o C, 1 atmosphere pressure) when contacted with pure oxygen (yO 2 =1). The deviation of the Bunsen coefficient from pure water (�0) can be calculated as the sum of positive and negative contributions of media components (Schumpe et al., 1981): �� 0 � log� � � � � � n � � organicxorganic � i�1 m w � K � �� H i j�1�k�1 k x j z 2 k � � � j (Eqn. 6) where Ki and Hi are experimentally determined coefficients for (n) different organic components such as sugar and ions of (m) dissociated salts. These coefficients are multiplied by the respective concentrations (xi) and also the valence squared in the case of ions. The contribution coefficients for some important plant culture media components are given in table 2A. Noting that � is roughly proportional to dissolved oxygen, and log(y) is positive for y>1, a positive value of Ki or Hi will decrease solubility. Nitrate ions (NO3 - ), therefore, reduce solubility, whereas ammonium (NH4 + ) will increase oxygen solubility. The equilibrium solubility of oxygen in several ‘typical’ plant culture media is presented as table 2B, demonstrating that the presence of nutrients generally suppresses oxygen solubility. This effect is relatively small (fresh MS medium has about 4% reduction), but the practice of utilizing high medium osmoticum (e.g. 5x sucrose) to ‘harden’ somatic embryos in preparation for transfer out of tissue culture conditions can result in a significantly reduced oxygen solubility. The Bunsen coefficient (and its counterpart the Henry’s Law coefficient) provides a means of quantitatively understanding the dissolved oxygen available to the micropropagule. The final and most obvious means of changing the equilibrium dissolved oxygen level is the alteration of the gas oxygen mole fraction. Air is composed of roughly 21% oxygen, and supplementation of plant tissue culture gas with up to 50-60 % oxygen is not usually inhibitory for plant tissues. Part of the reason for the lack of toxicity at even higher oxygen levels, is that the tissue does not actually experience these levels of oxygen
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 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
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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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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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Liquid Culture Systems for in vitro Plant Propagation

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