Protocols for Micropropagation of Woody Trees and Fruits

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MICROPROPAGATION OF BLACK LOCUST 195 Figure 1. Micropropagation of black locust (Robinia pseudoaoacia L.). (A) Mother plants with 35 year age. (B) Explant on initiation medium. (C) Sprouting of axillary bud. (D) Shoot elongation and multiplication. (E) Rooting culture. (F) Plant in nutritious plate plug, five weeks after transplantation. Courtesy of Professor Xiaofang Luo. The pH of the medium was adjusted to 5.8 using 1M NaOH or HCl. Agar gel strength was 1200 g/cm 2 . The agar was dissolved by heating in a microwave oven (2 min for each 250 ml media in a 1000-W microwave oven) and 10 ml was dispensed in each glass tube (25 × 12.5 mm) for initiation culture and 40 ml into each culture vessel (250 cm 3 in size) for subcultures, respectively. The glass tubes or vessels were 2 sealed with whorl plastic cover, marked and autoclaved at 121° C (1.05 kg/cm ) for 20 min.
196 J. ZHANG ET AL. 2.2.2. Culture Conditions Initiation, multiplication and root induction on black locust cultures were achieved at 26 ± 1° C under a 16-h photoperiod with fluorescent illumination (TLD 36W/33, –2 –1 OSRAM, Germany) at a light intensity of 36 µmol m s . The growth of cultures declined at a temperature below 23° C, and hyperhydricity occurred at 30° C. The leaves of the explants crumpled and the caudexes became transparent and curved. In some of the explants, the hyperhydricity could be recovered to normal growth by transferring them onto the fresh culture medium, and maintained at the room temperature for more than 60 days. Table 1. Composition of culture media for black locust. Compounds Initiation medium Proliferation medium Rooting medium MS medium Full strength Full strength Half strength Sucrose (g/L) 30 20–30 Agar (g/L) 5 5 NAA (mg/L) 0.1 1 6-BA (mg/L) 0.5 1 IBA (mg/L) — — pH 5.8 5.8 a 20 5 0.2 — 0.1 5.8 a Half strength means the concentration of major element salts is halved. NAA: naphthalene acetic acid; 6-BA: 6-benzylamino-purine; IBA: 3-indolebutyric acid. 2.3. Initiation of Shoot Cultures The sterilized explants were grown on the initiation culture medium for about 5 to 10 days until sprouting. The contaminated cultures were discarded after autoclaving 2 at 121°C (1.05 kg/cm ) for 20 min. It was important to take care of the contaminated cultures. In general, the number of contaminated explants was low, indicating the selected sterilizing agent was effective. Too high concentrations of the sterilizing agent were injurious to both contaminating microorganisms and plant material. At lower concentrations, the contamination rate of plant material was very high. Generally, 50% of decontamination could be achieved by optimizing the sterilization procedure without damaging plant material seriously. 2.4. Elongation and Multiplication of Shoots Shoots derived from the initial shoot cultures were cut into 1.5 cm long segments which were cultured on the proliferation culture medium (Table 1, Figure 1D). Usually 4–5 segments were placed in each culture vessel. After 4–5 days, shoots from the adventitious buds grew fast throughout the following 35 days. To keep the cultures in vigorous growth, the subcultures were carried out on the fresh culture medium, at a every 28-day interval.
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PROTOCOLS FOR MICROPROPAGATION OF W
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A C.I.P. Catalogue record for this
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vi TABLE OF CONTENTS 14. Root induc
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viii TABLE OF CONTENTS 43. Micropro
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x PREFACE on precise stepwise proto
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CHAPTER 1 TOTIPOTENCY AND THE CELL
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TOTIPOTENCY AND THE CELL CYCLE 5 a
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2.2. Plant Bioregulators and the Ce
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TOTIPOTENCY AND THE CELL CYCLE 9 in
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TOTIPOTENCY AND THE CELL CYCLE 11 F
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Bartel, D. (2004) MicroRNAs: genomi
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CHAPTER 2 MICROPROPAGATION VIA ORGA
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MICROPROPAGATION OF SLASH PINE Tabl
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MICROPROPAGATION OF SLASH PINE Amon
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2.6. Field Testing MICROPROPAGATION
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CHAPTER 3 MICROPROPAGATION OF COAST
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MICROPROPAGATION OF SEQUOIA SEMPERV
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CHAPTER 4 MICROPROPAGATION OF PINUS
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MICROPROPAGATION OF PINUS PINEA L.
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42 2.1. Organ Culture K. ISHII ET A
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44 K. ISHII ET AL. scent light, 70
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46 Chemicals (mg/l) K. ISHII ET AL.
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48 Table 3. Effects of plant growth
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50 K. ISHII ET AL. Gupta, P.K. & Du
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52 C. HARGREAVES AND M. MENZIES (Me
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54 C. HARGREAVES AND M. MENZIES Con
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56 2.1.3. Organogenesis from Field-
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58 C. HARGREAVES AND M. MENZIES Fig
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60 C. HARGREAVES AND M. MENZIES Fig
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62 C. HARGREAVES AND M. MENZIES For
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64 C. HARGREAVES AND M. MENZIES and
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CHAPTER 7 GENETIC FIDELITY ANALYSES
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PLANT GENETIC FIDELITY 69 Plant mat
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e added to samples, as this fluoroc
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11. Determine the mean channel numb
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3. In addition to testing various b
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GeneScan internal size standards: 4
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3500 3000 2500 2000 1500 1000 500 0
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PLANT GENETIC FIDELITY 81 microprop
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PLANT GENETIC FIDELITY 83 Steinkell
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86 2.1. Explant Preparation M.G. OS
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88 WPM (Lloyd & McCown, 1980) Macro
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90 M.G. OSTROLUCKÁ ET AL. on the m
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CHAPTER 9 MICROPROPAGATION OF MEDIT
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2.1. Explant Preparation MICROPROPA
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MICROPROPAGATION OF MEDITERRANEAN C
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108 D.T. NHUT ET AL. Larue (1953) a
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110 D.T. NHUT ET AL. HgCl2 added wi
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112 2.4. Establishment of Shoot Cul
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114 D.T. NHUT ET AL. Table 5. Effec
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116 D.T. NHUT ET AL. culture to be
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118 D. EWALD ability of the plant m
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120 D. EWALD Figure 1. Yew micropro
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122 D. EWALD Root development. Afte
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CHAPTER 12 MICROPROPAGATION OF LARI
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MICROPROPAGATION OF LARIX SPECIES V
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CHAPTER 13 PROPAGATION OF SELECTED
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PROPAGATION OF SELECTED PINUS GENOT
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Page 148 and 149: PROPAGATION OF SELECTED PINUS GENOT
Page 150 and 151: CHAPTER 14 ROOT INDUCTION OF PINUS
Page 152 and 153: ROOT INDUCTION OF PINUS SYLVESTRIS
Page 154 and 155: ROOT INDUCTION OF PINUS SYLVESTRIS
Page 156 and 157: CHAPTER 15 MICROPROPAGATION OF BETU
Page 158 and 159: MICROPROPAGATION OF BETULA PENDULA
Page 166 and 167: CHAPTER 16 PROTOCOL FOR DOUBLED-HAP
Page 168 and 169: DOUBLED-HAPLOID MICROPROPAGATION IN
Page 182 and 183: CHAPTER 17 IN VITRO PROPAGATION OF
Page 184 and 185: IN VITRO PROPAGATION OF FRAXINUS SP
Page 188 and 189: Axillary shoots (number) IN VITRO P
Page 196 and 197: CHAPTER 18 MICROPROPAGATION OF BLAC
Page 200 and 201: 2.5. Rooting and Acclimatization MI
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Page 204 and 205: 202 V. RAJESWARI AND K. PALIWAL tha
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Page 208 and 209: 206 V. RAJESWARI AND K. PALIWAL 2.3
Page 210 and 211: 208 V. RAJESWARI AND K. PALIWAL Fig
Page 212 and 213: 210 2.8. Hardening V. RAJESWARI AND
Page 214 and 215: CHAPTER 20 MICROPROPAGATION OF SALI
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Page 242 and 243: 242 2.4.1. Rooting of Apical and Ba
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CHAPTER 23 MICROGRAFTING IN GRAPEVI
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MICROGRAFTING IN GRAPEVINE 251 and
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MICROGRAFTING IN GRAPEVINE 253 Tabl
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2.4. Culture Conditions MICROGRAFTI
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MICROGRAFTING IN GRAPEVINE 257 Feuc
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CHAPTER 24 MICROGRAFTING GRAPEVINE
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MICROGRAFTING GRAPEVINE FOR VIRUS I
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CHAPTER 25 APRICOT MICROPROPAGATION
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APRICOT MICROPROPAGATION Figure 1.
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APRICOT MICROPROPAGATION Figure 2.
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APRICOT MICROPROPAGATION 2.2.2. Hyp
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2.4. Acclimatization APRICOT MICROP
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APRICOT MICROPROPAGATION occurs fre
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CHAPTER 26 IN VITRO CONSERVATION AN
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IN VITRO CONSERVATION AND MICROPROP
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CHAPTER 27 MICROGRAFTING OF PISTACH
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MICROGRAFTING OF PISTACHIO Constitu
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MICROGRAFTING OF PISTACHIO 2.2.2. C
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MICROGRAFTING OF PISTACHIO 6. Incub
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MICROGRAFTING OF PISTACHIO 9. The r
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CHAPTER 28 PROTOCOL FOR MICROPROPAG
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MICROPROPAGATION OF CASTANEA SATIVA
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CHAPTER 29 MICROPROPAGATION OF CASH
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MICROPROPAGATION OF CASHEW 2.2.1. E
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MICROPROPAGATION OF CASHEW axillary
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MICROPROPAGATION OF CASHEW Table 1.
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MICROPROPAGATION OF CASHEW shade an
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CHAPTER 30 IN VITRO MUTAGENESIS AND
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IN VITRO MUTAGENESIS AND MUTANT MUL
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336 R.I. IYER compounds (Iyer et al
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A 338 2.2. Culture Medium R.I. IYER
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340 R.I. IYER Figure 2. Formation o
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342 R.I. IYER Figure 3. Formation o
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344 R.I. IYER Rao, Y.S., Mathew, K.
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346 B.K. BISWAS AND S.C. GUPTA marg
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348 B.K. BISWAS AND S.C. GUPTA (iv)
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350 B.K. BISWAS AND S.C. GUPTA 2.4.
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352 B.K. BISWAS AND S.C. GUPTA Figu
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354 B.K. BISWAS AND S.C. GUPTA tree
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356 B.K. BISWAS AND S.C. GUPTA Figu
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358 B.K. BISWAS AND S.C. GUPTA Drew
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CHAPTER 33 MICROPROPAGATION PROTOCO
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MICROPROPAGATION FOR MICROSPORE EMB
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374 L. TIAN AND S.I. SIBBALD younge
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376 L. TIAN AND S.I. SIBBALD Table
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378 L. TIAN AND S.I. SIBBALD 2.4. R
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CHAPTER 35 MICROPROPAGATION OF JUGL
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MICROPROPAGATION OF JUGLANS REGIA L
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CHAPTER 36 TISSUE CULTURE PROPAGATI
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PROPAGATION OF MONGOLIAN CHERRY AND
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410 M. PASQUAL AND E.A. FERREIRA 2.
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414 M. PASQUAL AND E.A. FERREIRA ch
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418 D.T. NHUT ET AL. its economical
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420 D.T. NHUT ET AL. Table 1. Shoot
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422 D.T. NHUT ET AL. Figure 3. Orga
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424 D.T. NHUT ET AL. mg l -1 NAA +
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426 D.T. NHUT ET AL. Nakasone, H.Y.
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428 C. LIU ET AL. C. cujete L. is c
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430 C. LIU ET AL. Table 1. Composit
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432 C. LIU ET AL. 6. Excise the sho
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434 Fresh weight per plantlet (mg)
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436 C. LIU ET AL. 4. REFERENCES Aga
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438 M. MISHRA ET AL. micropropagati
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440 M. MISHRA ET AL. each plate ino
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Section C
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446 M.G. OSTROLUCKÁ ET AL. from me
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448 M.G. OSTROLUCKÁ ET AL. regener
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450 M.G. OSTROLUCKÁ ET AL. Figure
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452 M.G. OSTROLUCKÁ ET AL. 2.6.3.
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454 counts counts M.G. OSTROLUCKÁ
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CHAPTER 42 PROTOCOL FOR MICROPROPAG
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AN medium (Anderson, 1980) MICROPRO
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MICROPROPAGATION OF VACCINIUM VITIS
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2.6. Flow Cytometry Analysis MICROP
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CHAPTER 43 MICROPROPAGATION OF BAMB
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BAMBOO MICROPROGATION BY AXILLARY S
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CHAPTER 44 IN VITRO CULTURE OF TREE
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IN VITRO CULTURE OF TREE PEONY 479
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500 M. MHATRE from various natural
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502 2.2. Disinfection of Plant Mate
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504 M. MHATRE soil in polythene bag
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506 M. MHATRE Figure 2. Pineapple,
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508 M. MHATRE Escalona, M., Lorenzo
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510 J.M. AL-KHAYRI Tunisia, Morocco
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512 J.M. AL-KHAYRI 2.1.2. Separatio
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514 2.2. Culture Medium J.M. AL-KHA
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516 J.M. AL-KHAYRI 5. Isolate callu
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518 J.M. AL-KHAYRI 3. Water the tra
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520 J.M. AL-KHAYRI expressed from c
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522 J.M. AL-KHAYRI alcohol and twic
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524 J.M. AL-KHAYRI potential. Simil
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526 J.M. AL-KHAYRI Barreveld, W.H.
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528 D.T. NHUT ET AL. arsenide chip
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530 D.T. NHUT ET AL. Figure 2. Plan
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532 D.T. NHUT ET AL. Figure 4. Plan
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534 D.T. NHUT ET AL. plantlets. The
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536 D.T. NHUT ET AL. Figure 11. Fre
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538 D.T. NHUT ET AL. Figure 13. Sub
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540 D.T. NHUT ET AL. The response o
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CHAPTER 48 IN VITRO MUTAGENESIS IN
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IN VITRO MUTAGENESIS IN BANANA 545
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3.3. Observations to be Recorded IN
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