Liquid Culture Systems for in vitro Plant Propagation

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530 Dirk Wilken et al. described a retention time of 3.8 min (�-citral) and 4.2 min (�-citral) was observed. 2.5.5 Sample preparation and determination of oleanolic acid Samples of approximately 190-210 mg of dry material were extracted with 90 ml dichloromethane in a Soxhlet apparatus for 30 min. The organic phase was filtered and taken to dryness under reduced pressure. Percentage recovery was estimated by spiking an inert support with a known amount of oleanolic acid. The oleanolic acid content of the samples was determined by HPLC using calibration curves as reported by Halkes (1998). A Merck- Hitachi (Darmstadt, F.R.G.) HPLC equipment with UV detector (Model Series L-4000, Column: LiChrocart 5 �m RP 18 Select B, 250 mm) and a data processor were used. The mobile phase consisted of acetonitrile, water and acetic acid (70:30:0.5; v/v), the flow rate was 1.0 ml min -1 . The signal was detected at 220 nm. The identity of oleanolic acid was checked by coinjection of a reference sample isolated from Fabiana. Under the working conditions described, the retention time for oleanolic acid was 14.1 – 14.5 min. 3. Results The multiplication rates after four weeks for shoot and callus cultures in microcontainers as well as for shoots in TIS are given in table 1. Although the increase of biomass differed considerably between the four species investigated, shoot growth was always highest in the TIS compared to culture in microcontainers. Multiplication rates of the callus cultures ranged from 3.1 within four weeks for Cymbopogon to 8.6 for Lavandula (Table 1). These callus cultures were used to establish cell suspensions which in turn were used as inoculum for Lavandula and Hypericum bioreactor cultures. The growth curves are given in figure 1. Contents of bioactive compounds for all in vitro cultures in comparison with plants grown in the field are shown in table 2. For Lavandula the content of rosmarinic acid was higher for shoots and cells grown in any tissue culture system compared to field grown plants (containing 4.8 mg rosmarinic acid per g dry weight). The highest concentration of rosmarinic acid (108 mg g -1 dry weight) was found in callus cultures. For Lavandula also the composition of the essential oil from field grown as well as from in vitro grown shoots was investigated (Figure 2). This analysis revealed that
Comparison of secondary plant metabolite production 531 Table 1: Multiplication rates (increase of biomass) of four plant species producing secondary metabolites in different tissue culture systems within a period of 4 weeks Shoots from microcontainers Lavandula Hypericum Cymbopogon Fabiana 2.8 3.5 9.0 3.2 Shoots from TIS 16.5 7.7 18.0 5.0 Callus culture 8.6 5.0 3.1 8.1 Table 2: Contents of bioactive compounds for in vitro cultures of four plant species in comparison to plants grown in the field (concentrations are given in mg g -1 dry weight) Species Lavandula Hypericum Cymbopogon Fabiana Compound rosmarinic acid hypericin �-citral �-citral oleanolic acid Shoots from microcontainers 55.3 0.15 0.27 0.46 0.01 Shoots from TIS 5.7 0.18 0.35 0.54 0.01 Callus culture 108 0.00 0.00 0.00 0.005 Suspension culture 50 0.00 0.00 0.00 0.004 Bioreactor culture 60 0.00 Field plant 4.8 0.52 3.95 10.25 0.28 Table 3: Concentration of bioactive compounds in plant material of Lavandula and Hypericum from TIS vessels. Material was fractionated according to the colour of the material: Depending on the position of the material within the vessel, the biomass was green (top and side layers), yellow (inner part of the vessel) or brown (bottom layer) Colour of plant material fractions Content of rosmarinic acid in Lavandula (mg g -1 dry weight) green (top layer) 7.5 green (side layer) 4.0 Content of hypericin in Hypericum (mg g -1 dry weight) 0.12 yellow 1.9 0.11 brown 0.0 0.23
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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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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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Page 478 and 479: 482 Hans R. Gislerød et al. Table
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Page 520 and 521: 526 Dirk Wilken et al. 1. Introduct
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Page 532 and 533: Chapter 40 Cultivation of root cult
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Page 540 and 541: Chapter 41 Optimisation of Panax gi
Page 542 and 543: Optimisation of Panax ginseng liqui
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Page 560 and 561: Production of taxanes 569 2. Materi
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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
Page 572 and 573: 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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