Liquid Culture Systems for in vitro Plant Propagation

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Cultivation of root cultures of Panax 543 Saponin content (mg/g dr.wt.) 22 20 18 16 14 12 10 8 6 4 2 0 3,0 2,5 2,0 1,5 1,0 0,5 0,0 Rg1,Re Rf Rb1 Rb2 Rc Rd Particular ginsenosides 4a 4b Saponin content (mg/g dr.wt.) Figure 4: Results from (4a) RITA ® – Temporary immersion system (4b) Rg1 Rf Rb1 Rb2 Rc Rd Particular ginsenosides 5a 5b Figure 5: Results from (5a) the “Applikon” Bioreactor (Applikon, Netherlands) with total volume of 3-litre (5b) Saponin content (mg/g dr.wt.) 8 7 6 5 4 3 2 1 0 Rg1,Re Rf Rb1 Rb2 Rc Rd Particular ginsenosides 6a 6b Figure 6: Results from (6a) 250 ml Erlenmeyer flasks on rotary shaker (125 rpm) (6b)
544 Tomáš Vanek et al. 2.3 Detection of ginsenosides by HPLC Adventitious roots were homogenized and extracted with methanol (7 ml g -1 fresh weight) for 5 days at room temperature. The sample was then filtered, and evaporated to dryness under vacuum. The residue of extract was redissolved in distilled water and partitioned with dietyl-ether, and twice in n-BuOH saturated with water. The n-BuOH layer was concentrated in vacuo to obtain the crude saponin fraction (Sanada, 1974). The n-BuOH soluble fraction was analyzed by HPLC for detection and quantification of ginsenosides Rb1, Rb2, Rc, Rd, Re, Rf and Rg1. HPLC analyses were performed using a system consisting of two high pressure pumps (DeltaChrom, SDS 020 a SDS 030) with a mixer (SunChrom GmbH) and PDA detector (JASCO, MD 1510); a stainless steel column (250 x 4 mm) packed by reverse phase Si-C18, 7 µm (Biospher); flow-rate 1 ml min -1 . The injection volume was set up at 20 µl in the autosampler (TSP, AS300). Eluents: (A) 15 % acetonitrile and water, (B) 100 % acetonitrile; Gradient elution profile: 0 – 40 min, 0 – 35 % B; 40 to 45 min, 35 % B. The peaks were monitored by UV detection at 203 nm (Soldati and Sticher, 1980; Pietta et al., 1986; Petersen and Palmqvist, 1990). Each ginsenoside was identified by comparison of retention time and UV spectra with authentic ginsenosides purchased from Carl Roth GmbH and Co., Germany. Ginsenoside content was expressed in mg g -1 of dry weight. Presence of ginsenosides was additionally confirmed by LC-MS. 3. Results and discussion We have monitored ginsenoside production in callus cultures previously (Langhansova et al., 2002). We found callus cultures to have a high proliferation rate and also high yielding. However, the production of saponins varied significantly during the year. The content ranged between 0 – 2 % dry weight and the saponin content in callus as well as in cell suspension cultures was limited to two major ginsenosides, Rb1 or Rg1. The full range of ginsenosides distribution analogous to roots of native plants was only detected in adventitious roots. Hence, in this study we have concentrated on adventitious root production in vitro. The proportion of particular ginsenosides compared with native material is almost identical after cultivation in the rocking TIS of our own construction (Figure 1a). This suggests that the rocking TIS is the optimal way of producing these adventitious roots in vitro. The range in protopanaxadiol (ginsenosides: Rb1, Rb2, Rc and Rd) and protopanaxatriol (ginsenosides: Re and Rg1) groups were similar also in RITA ® TIS
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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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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
Page 534 and 535: Cultivation of root cultures of Pan
Page 538 and 539: Cultivation of root cultures of Pan
Page 540 and 541: Chapter 41 Optimisation of Panax gi
Page 542 and 543: Optimisation of Panax ginseng liqui
Page 550 and 551: 558 S. M. Nalawade et al. performan
Page 552 and 553: 560 S. M. Nalawade et al. Plantlets
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Page 556 and 557: 564 S. M. Nalawade et al. roots (Fi
Page 558 and 559: Chapter 43 Production of taxanes in
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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