Protocols for Micropropagation of Woody Trees and Fruits

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52 C. HARGREAVES AND M. MENZIES (Menzies et al., 1985; Menzies & Aimers-Halliday, 1997). More recent advances in the area of juvenility maintenance include the development of cryogenic methods in combination with somatic embryogenesis (Hargreaves & Smith, 1992, 1994; Hargreaves et al., 2002). Methods for organogenic material have been more difficult to develop though significant progress has been made with zygotic embryos and recently, shoot tip meristems (Hargreaves et al., 2004, 2005). Somatic embryogenesis was originally seen as the way forward in terms of delivering clonal forestry strategies to commercial forest operators, because the technique results in tissue that can be easily cryopreserved (thus ensuring juvenility while field testing takes place) and has the potential to produce millions of plants quickly (Durzan & Gupta, 1988; Attree & Fowke, 1991). However, major problems have been low genotype capture and a high labour cost of in vitro procedures (Park et al., 1998; Timmis, 1998). Genotype capture using organogenesis methods with zygotic embryos (mature and immature) or seedling/stool bed material is high and the in vitro culture period short. Media formulations include few plant growth regulators, and less skilled observation through developmental stages is required than with somatic embryogenesis (Hargreaves et al., 2005). Propagation techniques for radiata pine which utilise organogenic approaches are showing great versatility for a range of applications, including arresting of juvenile growth via cool storage and more recently developed cryogenic methods. These also extend to the preservation of genetic resources in the case where native populations may be under threat from human interference, including climate change and disease. The potential is promising for amplifying valuable seed that may be both limited in supply and less fit, in the case of historic seed collections and where new and novel hybrids may have been bred (Hargreaves et al., 2007). Other uses of the approach are to facilitate the safe (contamination free, pre-screened) dissemination of proven/novel genetic material internationally as shoot cultures. This material arrives in a ready-to-amplify condition. This chapter sets out to describe the current methods for organogenic multiplication and cryogenic storage of radiata pine. Details are also given for the transition of material from the laboratory to the nursery and field. 2.1. Explant Preparation 2.1.0. Culture Media Constituent name Constituent formula 2. EXPERIMENTAL PROTOCOL Shoot initiation Shoot multiplication Medium* Medium* (1/2 LP5) (LPch) Root initiation Medium** (GD) Major elements mg l –1 mg l –1 mg l –1 Ammonium sulphate (NH4)2SO4 200.00 Ammonium nitrate NH4NO3 200.00 400.00
ORGANOGENESIS AND CRYOPRESERVATION OF RADIATA PINE 53 Constituent name Constituent formula Shoot initiation Shoot multiplication Root initiation Calcium nitrate Magnesium sulphate Ca(NO3)2 × 4H2O MgSO4 × 7H2O 600.00 180.00 1200.00 360.00 250.00 Potassium chloride KCl 300.00 Potassium dihydrogen orthophosphate KH2PO4 135.00 270.00 Potassium nitrate Sodium dihydrogen orthophosphate KNO3 NaH2PO4 × 2H2O 900.00 1800.00 1000.00 100.00 Disodium hydrogen orthophosphate Dodecahydrate Na2 HPO4 × 12H 2O 75.00 Minor elements mg l –1 mg l –1 mg l –1 Manganous sulphate MnSO4 × 4H2O 5.0000 10.000 20.00 Boric scid H3BO3 3.1000 6.200 5.00 Zinc sulphate ZnSO4 × 7H2O 4.3000 8.600 1.00 Potassium iodide KI 0.0400 0.080 1.00 Cupric sulphate CuSO4 × 5H2O 0.1250 0.250 0.20 Sodium molybdate Na2MoO4 × 2H2O 0.1250 0.250 0.20 Cobaltous chloride CoCl2 × 6H2O 0.0125 0.025 0.20 Calcium mg l –1 Calcium chloride Dihydrate CaCl2 × 2H2O 150.00 Vitamins mg l C12H17CIN4OS × HCl 0.200 0.400 5.00 Nicotinic acid Pyridoxine HCl C6H5NO2 C8H11NO3 × HCl 5.00 0.50 –1 Thiamine HCl Iron mg l –1 mg l –1 mg l –1 Ethylenediaminetetraacetic acid Na2EDTA 15.00 30.00 30.00 Ferrous sulphate FeSO4 × 7H2O 20.00 40.00 40.00 NAA mg l –1 1-Naphtalene acetic acid (NAA) C12H10O2 0.50 IBA mg l –1 Indole-3-butyric acid (IBA) C12H13NO2 1.00
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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
Page 10 and 11: x PREFACE on precise stepwise proto
Page 12 and 13: CHAPTER 1 TOTIPOTENCY AND THE CELL
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Page 16 and 17: 2.2. Plant Bioregulators and the Ce
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Page 30 and 31: 2.6. Field Testing MICROPROPAGATION
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Page 34 and 35: MICROPROPAGATION OF SEQUOIA SEMPERV
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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
Page 58 and 59: 50 K. ISHII ET AL. Gupta, P.K. & Du
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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
Page 76 and 77: PLANT GENETIC FIDELITY 69 Plant mat
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Page 88 and 89: PLANT GENETIC FIDELITY 81 microprop
Page 90 and 91: PLANT GENETIC FIDELITY 83 Steinkell
Page 92 and 93: 86 2.1. Explant Preparation M.G. OS
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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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Protocols for Micropropagation of Woody Trees and Fruits

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