Protocols for Micropropagation of Woody Trees and Fruits

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440 M. MISHRA ET AL. each plate inoculate 10 zygotic embryos. Incubate cultures at 25°C ± 2°C in the dark for 4 weeks. Globular embryos appeared after 4 weeks of inoculation, which will form bipolar-shape. Subculture somatic embryos on this medium (after 4 weeks), this will result in proliferation of embryos (Figure 2 a, b, c). 2.3.2. Somatic Embryo Maturation and Germination After 4 weeks, several creamy white to light yellow thread like embryos will appear. Transfer the clumps of somatic embryos on half strength MS medium containing 3% sucrose and 0.8% Difco bacto-agar, 400 mg/l glutamine, MS vitamins, 0.2 mg/l BAP and 0.1 mg/l NAA (Regeneration medium). (Table 1) Incubate the cultures under 16 h light (25 µmol m –2 s –1 ) at 25°C ± 2°C. Allow somatic embryos to germinate. If callus formation starts at the base of the shoots, than transfer the cultures on MS medium devoid of any plant growth regulators. This will check the callus growth at the base of shoot. Figure 2. Somatic embryogenesis in papaya. A) Immature zygotic embryo, B) Somatic embryo, C) Germination of embryos, D) Rooting of plantlets, E) Acclimatization of plantlets, F) Acclimatized plants in poly bags, G) Establishment of plants in field. 2.3.3. Rooting and Acclimatization Transfer regenerated plantlets (5 cm) in transparent culture bottles containing autoclaved vermiculite supplemented with ¼ MS liquid medium, and incubate them under light (25 µmol m –2 s –1 ) illumination) at 25ºC ± 2ºC for 2–3 weeks. Sometimes good rooting is a problem. In such case, add 1 mg/l IBA solution in vermiculite for better root growth. Gradually loosen the cap of bottle as soon as plant starts touching the cap of bottle. Transfer the plantlets into a poly house provided with 50% shade
MICROPROPAGATION OF PAPAYA 441 net and misting. Once plant develops 4–6 fully opened leaves, transfer them into poly bags filled with Soil + Sand + Farm Yard Manure (1:1:1) (Figure 2). 2.3.4. Somatic Embryogenesis from Hypocotyls Select 10-day-old in vitro grown seedling. Cut the hypocotyl sections and inoculate them on half strength MS (Induction) medium. Incubate the cultures in the darkness for 4 weeks at 25°C ± 2°C for callus induction and subsequently somatic embryo formation. For embryo proliferation, germination, rooting and acclimatization follow the above mentioned protocol (2.3.1 to 2.3.3). 3. CONCLUSION Papaya is an important cash fruit crop owing to its fast maturation rate and continuous fruit production in subtropical and tropical climates. There are reports on micropropagation of papaya from mature field grown trees. However, the technique could not be reproduced for mass cloning of papaya at commercial scale. Latex oozing, in borne contamination, and poor in vitro rhizogenesis are some of the bottlenecks in micropropagation of papaya. 4. REFERENCES Ali, A.M. & Hogan, L. (1976) Tissue culture of Carica papaya. Hort. Sci. 11, 27. Arora, I.K. & Singh, R.N. (1978) In vitro plant regeneration in Papaya. Curr. Sci. 47, 839–883. Drew, R.A. (1988) Rapid clonal propagation of papaya in vitro from mature field grown Trees. Hort Sci. 23, 609–611. Drew, R.A. (1992) Improved techniques for in vitro propagation and germplasm storage of papaya. Hort Sci. 27, 1122–1124. Fitch, M.M. (1993) High frequency somatic embryogenesis and plant regeneration from Papaya hypocotyl callus. Plant Cell Tiss. Org. Cult. 32, 205–212. Fitch, M.M. & Manshardt, R.M. (1990) Somatic embryogenesis and plant regeneration from immature zygotic embryos of Papaya (Carica papaya). Plant Cell Rep. 9, 320–324. Litz, R.E. & Conover, R.A. (1978) Tissue culture propagation of Papaya. Proc. Fla. State Hort. Soc. 90, 245–246. Litz, R.E., O ’ ’ Hair, S.K. & Conover, R.A. 1983. In vitro growth of Carica papaya L cotyledons. Sci. Hort. 19: 287–293. Murashige, T. & Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15, 473–497. Rajeevan, M.S. & Pandey, R.M. (1986) Lateral bud culture (Carica papaya) for clonal propagation. Plant Tiss. Org. Cult. 6, 181–188. Sampson, J.A. (1986) Tropical Fruits. Second Ed. Longman Scientific and Technical, Longman Inc., New York, NY. Van Droogenbroeck, B., Kyndt, T., Martens, I., Romeijn-Peeters, E., Scheldeman, X., Romero-Motochi, J.P., Van Dammep, G.P. & Gheysen, G. (2003) Phylogenetic analysis of the highlands papayas (Vasconcellea) and allied genera (Caricaceae) using PCR-RFLP. Theor. Appl. Genet. 108, 1473–1486.
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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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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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IN VITRO CULTURE OF TREE PEONY 493
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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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