Protocols for Micropropagation of Woody Trees and Fruits

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MICROPROPAGATION OF SEQUOIA SEMPERVIRENS 3. Shoot elongation and spontaneous rooting: Healthy shoots with healthy needles are cut into 1 cm and culture in the jars containing ½ WS without PGR for 8 weeks. Spontaneous rooted shoots (about 20 to 30%) are transferred to the greenhouse for further shoot elongation and the rest of shoots can be segmented for in vitro proliferation (repeat step 2). Elongation of shoots can be achieved when shoots are grown on WS basal medium only, however adding activated charcoal can help promote shoot elongation. Stage 1: Establishment of explants Stage 2: Shoot proliferation Stage 3: Shoot elongation and spontaneous rooting Spontaneous rooted shoots Greenhouse or field-grown plants of Sequoia Sempervirens Cut stem into 10 cm segments and sterilize with Clorox (10%) Culture stem segments vertically in test tubes containing WS medium w/o PGR Excise axillary shoots from stem segments Culture vertically in jars containing WS medium with zeatin (15 µM) Excise axillary shoots from stem segments Cut into 1 cm sections with 3 to 5 axillary buds Cut into 1 cm sections, each with 3 to 5 axillary buds Culture vertically in jars containing WS medium Greenhouse for acclimatization Subculture for further shoot proliferation Discard contaminated stem segments Discard contaminated stem segments Repeat shoot proliferation 4 weeks 4 weeks 8 weeks Figure 1. A schematic diagram of the overall micropropagation protocol for S. sempervirens using nodal stem segments. 27
28 S.S. KORBAN AND I.-W. SUL Seed. Following surface-sterilization, testae are removed, and a thin layer of axenic female gametophytes are excised from the embryo (Ball, 1987). These are introduced into petri plates containing a modified Murashige & Skoog (MS) medium containing 2 µM BA and 2 µM kinetin. Within 2 months, organogenic callus is formed, and following monthly subcultures, shoots are observed. Nodal stem segments. Each nodal stem explant is placed into a test tube containing Wolter & Skoog (WS) (1966) basal medium without any PGRs. Contaminated explants are discarded, and elongated healthy axillary shoots are excised, and cultured on a WS basal medium for further establishment as descrybed below. Figure 2. Micropropagation of S. sempervirens. Using greenhouse-grown mother plants as sources of nodal stem segments (A), these are then introduced in vitro, established, and proli- ferated (B). Following shoot proliferation medium, these are transferred to a fresh medium to elongate (C), and root spontaneously (D). Shoot buds (E, F) as well as somatic embryos (G, H) can also be induced on in vitro-grown needles.
Page 2 and 3: PROTOCOLS FOR MICROPROPAGATION OF W
Page 4 and 5: A C.I.P. Catalogue record for this
Page 6 and 7: vi TABLE OF CONTENTS 14. Root induc
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Page 10 and 11: x PREFACE on precise stepwise proto
Page 12 and 13: CHAPTER 1 TOTIPOTENCY AND THE CELL
Page 14 and 15: TOTIPOTENCY AND THE CELL CYCLE 5 a
Page 16 and 17: 2.2. Plant Bioregulators and the Ce
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Page 20 and 21: TOTIPOTENCY AND THE CELL CYCLE 11 F
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Page 24 and 25: CHAPTER 2 MICROPROPAGATION VIA ORGA
Page 26 and 27: MICROPROPAGATION OF SLASH PINE Tabl
Page 28 and 29: MICROPROPAGATION OF SLASH PINE Amon
Page 30 and 31: 2.6. Field Testing MICROPROPAGATION
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Page 34 and 35: MICROPROPAGATION OF SEQUOIA SEMPERV
Page 42 and 43: CHAPTER 4 MICROPROPAGATION OF PINUS
Page 44 and 45: MICROPROPAGATION OF PINUS PINEA L.
Page 50 and 51: 42 2.1. Organ Culture K. ISHII ET A
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Page 54 and 55: 46 Chemicals (mg/l) K. ISHII ET AL.
Page 56 and 57: 48 Table 3. Effects of plant growth
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Page 60 and 61: 52 C. HARGREAVES AND M. MENZIES (Me
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Page 64 and 65: 56 2.1.3. Organogenesis from Field-
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Page 74 and 75: CHAPTER 7 GENETIC FIDELITY ANALYSES
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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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CHAPTER 14 ROOT INDUCTION OF PINUS
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ROOT INDUCTION OF PINUS SYLVESTRIS
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ROOT INDUCTION OF PINUS SYLVESTRIS
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CHAPTER 15 MICROPROPAGATION OF BETU
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MICROPROPAGATION OF BETULA PENDULA
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CHAPTER 16 PROTOCOL FOR DOUBLED-HAP
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DOUBLED-HAPLOID MICROPROPAGATION IN
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CHAPTER 17 IN VITRO PROPAGATION OF
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IN VITRO PROPAGATION OF FRAXINUS SP
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Axillary shoots (number) IN VITRO P
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CHAPTER 18 MICROPROPAGATION OF BLAC
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MICROPROPAGATION OF BLACK LOCUST 19
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2.5. Rooting and Acclimatization MI
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MICROPROPAGATION OF BLACK LOCUST 19
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202 V. RAJESWARI AND K. PALIWAL tha
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204 V. RAJESWARI AND K. PALIWAL Mak
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206 V. RAJESWARI AND K. PALIWAL 2.3
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208 V. RAJESWARI AND K. PALIWAL Fig
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210 2.8. Hardening V. RAJESWARI AND
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CHAPTER 20 MICROPROPAGATION OF SALI
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MICROPROPAGATION OF SALIX CAPREA L.
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CHAPTER 21 MICROPROPAGATION OF CEDR
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2.1. Establishment of Shoot Culture
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MICROPROPAGATION OF CEDRELA FISSILI
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238 programmes is somatic embryogen
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240 Figure 2. A) Induction of organ
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242 2.4.1. Rooting of Apical and Ba
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244 of donor individuals and/or by
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246 J. MALÀ ET AL. Karnosky, D.F.,
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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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