Liquid Culture Systems for in vitro Plant Propagation

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Chapter 43 Production of taxanes in callus and suspension cultures of Taxus baccata L. K.Bru�áková, Z. Babincová & E.�ellárová Institute of Biology and Ecology, Faculty of Science, P. J. Šafárik University, Mánesova 23, 041 54 Košice, Slovakia. E-mail: brunakov@kosice.upjs.sk Abstract: The selection and cloning of cell lines for the establishment of long-term cell cultures of Taxus baccata L. is important for the prospective biotechnological production of taxanes. Callus cultures were established from young stems of adult trees and aseptically grown seedlings. All parts of young seedlings were more suitable for cell proliferation on callus-induction media as compared to young stems of adult trees. The best growing calli successfully achieved an average 8-fold increase in fresh weight after 20 months of culture. The growth characteristics of two seedling-derived cell lines were determined. The best growing calli were used for establishment of suspension cultures. Gamborg’s B5 medium supplemented with 3 mg l -1 2,4-dichlorophenoxyacetic acid, 0.5 mg l -1 kinetin and 1.5% polyvinylpyrrolidone, a phenolic-binding agent, was used as agar-solidified and liquid medium. A 20-month-old callus culture of T. baccata (VI/Ha) produced paclitaxel up to 0.0109 ± 0.0037 % of extracted dry weight basis. The content of taxanes was determined by high performance liquid chromatography or a competitive inhibition enzyme immunoassay system (CIEIA). A kinetic study of callus growth and taxane production was performed. Key words: Cell culture, kinetics study, taxane, paclitaxel, yew Abbreviations: E.D.W. – extracted dry weight; V/Kle – callus culture derived from cotyledon of an embryo-derived seedling (genotype V); VI/Ha – callus culture derived form hypocotyl of an embryo-derived seedling (genotype VI); GI – growth rate; PVP - polyvinylpyrrolidone 1. Introduction Plant cell culture is an alternative method for commercial propagation as well as production of secondary metabolites in vitro (George and Sherrington, 1984; Barz and Ellis, 1981). Paclitaxel (TAXOL ® ; Bristol-Myers Squibb Co.) (Figure 1) and docetaxel (TAXOTERE ® ; Rhône-Poulenc Rorer 567 A.K. Hvoslef-Eide and W. Preil (eds.), Liquid Culture Systems for in vitro Plant Propagation, 567–574. © 2005 Springer. Printed in the Netherlands.
568 K.Bru�áková et al. R O NH OH O O AcO HO OH OAc PhOCO Figure 1: The structure of paclitaxel and cephalomannine; paclitaxel: R-C 6H 10; cephalomannine: R-C 4H7 Co.) are derived from natural sources. At least 40 % of the phytopharmaceuticals being currently used in Western countries is of natural origin (Rout et al., 2000). With a unique mechanism of action – the inhibition of microtubule depolymerization – taxanes are considered as the most important drug in the treatment of breast carcinoma since antracyclines (Wagnerová and Andrašina, 1996). In 1992, TAXOL ® was approved by the United States Food and Drug Administration for treating ovarian and breast cancer. The history of paclitaxel, the chemistry and biosynthesis as well as mechanism of its action and broad antitumour spectra are well documented (Heinstein and Chang, 1994; Jennewein and Croteau, 2001). The enormous requirements for supplying TAXOL ®, originally derived from the bark of the Pacific yew, occurred as a result of TAXOL ® effectiveness in clinical trials of cancer treatment and can lead to devastation of natural yew resources (Song and Dumais, 1991). Plant cell cultures of Taxus sp. seem to be promising as a potential candidate for commercial TAXOL ® production in the near future. While the early work was concentrated on establishing cell lines and taxoid assay (Zhong, 1995; Wickremesinhe and Arteca, 1993), on selection of high-paclitaxel-producing cell and protoplast cultures for efficient paclitaxel production (Aoyagi et al., 2002), recently the emergence of recombinant DNA techniques has opened new possibilities of studying and modifying expression of genes related to taxane biosynthesis (Jennewein and Croteau, 2001). To achieve higher productivity, the selection of cell lines with appropriate genetic, biochemical and physiological properties is necessary. O H O
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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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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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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 552 and 553: 560 S. M. Nalawade et al. Plantlets
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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
Page 572 and 573: 582 Contents bubble aerated 165 bub
Page 574 and 575: 584 Contents ginsenoside content, p
Page 576 and 577: 586 Contents plant growth regulator
Page 578: 588 Contents -free microcorms 71 -f
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