Liquid Culture Systems for in vitro Plant Propagation

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246 C. Damiano et al. 2.2 Temporary immersion apparatus The TIS was composed of two glass bottles connected with a silicone tube; one of the bottles contained the explants, the other one contained the culture medium (Figure 1A). The medium was periodically transferred from one bottle to the other by pressure created by an air pump; the air introduced into the bottle was sterilised with a 0.2 �m filter. Glass beads on the bottom of the bottle containing plants were used to isolate the plants from the thin layer of liquid medium remaining in the bottle during the period without immersion and for a better regulation of gravitropism. 2.3 Experimental plan For each treatment 10 single shoots were used and 3 replications were carried out. After 60 days of culture, the multiplication rate (calculated as number of new shoots produced per cluster), fresh (FW) and dry weight (DW), obtained after dehydration in the oven at 110°C for 24 hrs, were recorded and the content of chlorophylls, carotenoids, and fructose (total or as sucrose) were measured. Water content (WC) percentage was calculated as WC = [(FW – DW)/FW] x 100. Percentage data were analysed after normalization according to the formula arc sin�% and the Standard Error was calculated. Data were analysed by analysis of variance. 2.4 Chlorophyll and carotenoids extraction and determination To analyse the a and b chlorophyll content, for each replication three samples of 100-200 mg of plant material from each treatment were prepared by soaking shoots in 12 ml of 80% acetone, according to Arnon (1949). Similarly samples for carotenoid extraction were prepared adding 50 mg of CaCO3, to prevent pigment oxidation, and using pure acetone, storing it overnight at –20°C, according to Jensen (1973). The pigment was spectrophotometrically determined (Spectrophotometer Hitachi 2000), measuring light absorbance of acetone extracts of chlorophylls (� = 663nm and 645nm, respectively for chlorophylls a and b) and carotenoids (� = 450nm). All amounts were expressed as mg g -1 of dry weight. 2.5 Fructose extraction and determination The fructose determination (total and as sucrose) was based on the estimation of a chromophoric substance formed by the reaction of resorcinol and fructose according to a modified Roe’s test (Davis and Gander, 1967).
Propagation of Prunus and Malus by temporary immersion 247 Three samples for each treatment from each replication of 5-10 mg of powdered dried tissue, were dissolved in water and after 5 minutes resorcinol (0.05 % in absolute ethanol) was added to the water solution. After addition of HCl (12 mol), the samples were heated at 77° C for 8 min and cooled immediately in ice for 5-10 min. Fructose content was determined with a spectrophotometer (Hitachi 2000) at � = 420nm. Since the test does not distinguish between free fructose and fructose bound in sucrose, estimation of the amount bound in the sucrose was performed after initial destruction of the free fructose in the sample by hot alkaline (KOH 2N) treatment. After KOH addition the samples were boiled for 10 min and cooled immediately in ice for 5-10 min. The difference between total fructose content and the fraction bound in sucrose molecules represented the content of free fructose. Table 1: Multiplication media Apple Peach Cherry Plum Macrosalts MS LP MS LP Microsalts MS MS MS LP Vitamins MS* MS MS MS Calcium pantothenate mgl -1 - - - 1 Biotin mg l -1 - - - 0.1 Riboflavin mg l -1 - - - 0.5 Sucrose g l -1 30 30 30 30 BA mg l -1 0.5 0.4 0.5 0.25 IBA mg l -1 0.1 0.06 0.1 0.1 GA3 mg l -1 - 0.03 - - Adenine sulphate. mg l -1 - 3 - - *Thiamine ten-fold concentrated (1mg l -1 ) MS: Murashige and Skoog, 1962; LP: Quoirin et al., 1977. Table 2: Multiplication rate of plants grown in different cultural conditions (Data are the mean of three values ± s.e.). L = liquid, S = solid, 30’ and 60’= TI period of 30 or 60 minutes Cultural conditions Plum Cherry Peach Apple L S 30’ 60’ 3 ± 0.32a 30 ± 2.89b n.r. 28 ± 2.54b 2 ± 0.31a 5 ± 1.05bc 6 ± 1.17b 4 ± 0.77c 3 ± 0.28a 20 ± 2.06b 8 ± 1.57c 14 ± 2.79d 3 ± 0.28a 10 ± 1.97b 2 ± 0.41c 8 ± 1.63b n.r.: not recorded Means within a column followed by different letter are significantly different (P � 0.05).
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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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Page 206 and 207: 198 Elio Jiménez González 1. Intr
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Page 210 and 211: 202 Elio Jiménez González Figure
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Page 216 and 217: 208 Elio Jiménez González germina
Page 218 and 219: 210 Elio Jiménez González Escalan
Page 220 and 221: Chapter 13 Use of growth retardants
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Page 232 and 233: Chapter 14 Somatic embryo germinati
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Page 258 and 259: Chapter 17 Optimisation of growing
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Page 268 and 269: 264 Katerina Grigoriadou et al. cul
Page 270 and 271: 266 Katerina Grigoriadou et al. 2.4
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Page 280 and 281: 276 Ch. Wawrosch et al. possesses s
Page 282 and 283: 278 Ch. Wawrosch et al. 3. Results
Page 284 and 285: 280 Ch. Wawrosch et al. References
Page 286 and 287: Chapter 20 Propagation of Norway sp
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Page 290 and 291: Propagation of Norway Spruce 287 su
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Page 300 and 301: 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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