Protocols for Micropropagation of Woody Trees and Fruits

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MICROGRAFTING IN GRAPEVINE 253 Table 1. Commonly used nutrient media for multiplication of grapevine shoots. Sl # Medium Macro elements a. b. 1 MS 1/2 1 3 NN 1 1 1 MS, Murashige & Skoog (1962) medium; 2 LM, Lloyd & McCown (1980) organics; 3 NN, Nitsch & Nitsch (1969) medium; 4 LS, Linsmaier & Skoog (1965) medium; 5 WPM, woody plant medium (Sigma catalogue, 1994); 6 B 5, Gamborg et al. (1968) medium; BAP, benzyl amino purine; IAA, indole-3-acetic acid; Ad. sul., adenine sulphate; NAA, naphthalene acetic acid; IBA, indole butyric acid. 2.3. In Vitro Grafting Micro elements mg l –1 Vitamins Growth adjuvant 2 LM-1 + inositol-2 BAP 0.5+IAA 0.08 4 LS-1 Thiamine 10 + Ad. sul. 40.53 + monobasic NaH 2PO4 218.4 + BAP 2.25 + NAA 0.09 c. 5 WPM 1 1 6 B5-1 Cal. Pan. 2 + monobasic NaH2PO4 168 + BAP 2.25 + IBA 0.5 30 d. MS 1 1 1 BAP 0.5 + IAA 0.2 20 e. MS 1/2 1/2 1 IAA 0.1 10 Carbon source g l –1 Reference 30 Pathirana & McKenzie, 2005 2.3.1. Using Shoot Apices In vitro micrografting can be attempted using shoot apices (0.3 mm) as explants, of the chosen rootstock and the scion to generate shoots on a large scale (Cantos et al., 1995). Once the rootstock culture is established as a rooted shoot, its top portion can be cut so that there is no nodal region left and a small cleft is made in the cut end. The shoot portion (bearing 3–4 buds) of the scion is sliced from the rest and inserted in the cleft of the rootstock. The entire set is again cultured in vitro on a medium that supports growth (Table 1). Bass et al. (1988) have used animal serum in the culture medium to facilitate the development of grapevine meristems (0.1 mm, 1–2 leaf primordial) and report an increase in the number of meristems developing into plantlets as well as in the rate of their in vitro differentiation. The healing of the callus occurs during the first 6 days and the establishment of vascular junction – between the 8 and 12th day after grafting. This is the critical period for successful micrografting and results in 60% survival of the micrografted plants (Cantos et al., 1995). 20 Mhatre et al., 2000
254 M. MHATRE AND V.A. BAPAT 2.3.2. Using in Vitro Produced Shoots Tissue culture generated rootstock rooted plantlets, grown until several leaves are produced (6–8 weeks) can be divided into 3–4 explants, each containing 1–2 nodes with leaves. The apex of the rootstock nodal explant is cut longitudinally with a sharp sterilized blade, producing a small (2–4 mm) longitudinal cleft leaving the rootstock decapitated. An appropriate scion with a single node and a leaf can be selected to match the size of the rootstock. The basal part of the stem of the scion should be cut in such a way that the wedge formed, matches the cleft of the rootstock. It is then fitted on to the cleft of the rootstock grown in the medium (Figure 2). The graft tissue is allowed to develop around the union during subsequent subculture. In this way, two grafts can be established per tissue culture vessel (Pathirana & McKenzie, 2005). After insertion of the scion into the rootstock cleft, wrapping in sterile aluminium foil to hold the graft can also be attempted to strengthen and support the graft (Tanne et al., 1993). However, to accomplish this is laborious and time consuming under aseptic conditions. In case the scion does not fit properly on the rootstock, dipping the lower end of the scion in the medium before fitting it helps to establish the graft. This is because of two reasons (i) the cut surfaces are kept moist until relative humidity of the tub/vessel is established and (ii) dipping ensures that the nutrients are directly supplied to the graft site (Pathirana & McKenzie, 2005). Figure 2. Diagrammatic representation of in vitro micrografting.
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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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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
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Page 242 and 243: 242 2.4.1. Rooting of Apical and Ba
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Page 266 and 267: CHAPTER 25 APRICOT MICROPROPAGATION
Page 268 and 269: APRICOT MICROPROPAGATION Figure 1.
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Page 278 and 279: CHAPTER 26 IN VITRO CONSERVATION AN
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Page 288 and 289: CHAPTER 27 MICROGRAFTING OF PISTACH
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Page 298 and 299: 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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