Protocols for Micropropagation of Woody Trees and Fruits

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PROPAGATION OF MONGOLIAN CHERRY AND NANKING CHERRY Table 1. Murashige and Skoog Minimal Organic (MSMO) medium (Murashige and Skoog, 1962). MS Macro Nutrients NH4NO3 KNO3 CaCl2 · 2H2O MgSO4 · 7H2O Compound Amount [mg l –1 ] KH2PO4 MS Micro Nutrients H3BO3 MnSO4 · H2O ZnSO4 · 7H2O KI Na2MoO4 · 2H2O CuSO4 · 5H2O CoCl2 · 6H2O Iron Sources FeSO4 · 7H2O Na2EDTA Vitamins Thiamine HCl Pyrodixin HCL Nicotinic acid 1650 1900 440 370 170 6.2 16.9 8.6 0.8 0.25 0.025 0.025 27.8 37.3 0.4 0.5 0.5 Organic constituents Inositol 100 Sucrose Varies Agar Varies 2.2.5. Shoot Regeneration and Maintenance As in other woody plant species, genetically uniform plants of Mongolian and Nanking cherry are obtained from cultures established from shoot tips, buds or meristems. Other organs and callus would provide a risk of inducing variability (Druart & Gruselle, 1986). Thus, suitable PGR combinations and concentrations have to be used to differentiate new axillary buds (rosettes) and to develop into shoots without callus formation. In our work, MSMO media were supplemented with combinations of various PGRs for initiation and establishment of cultures. The optimal combinations for culture establishment and performance have been determined following a number of experiments and are presented in Table 2. The combination of IBA and/or NAA with 8.9 µM BA is the best for initiation of both Mongolian and Nanking cherry cultures. The number of rosettes produced on media with 8.9 µM BA is significantly higher than on media with 4.4 µM BA. Initially, one explant (dormant bud) was placed on medium to establish the culture. Following 4 weeks in culture, approx. 90% explants survived. Most of them produced a single rosette and a few produced two rosettes (out of 30 explants). For Mongolian cherry media supplemented with lower concentration of BA (4.4 µM) 395
396 K. PRUSKI can also be used (Table 2). Prediction of the number of rosettes per explants is an important element in multiplication scheduling for both Mongolian and Nanking cherry cultures. Table 2. Effects of growth regulators on culture initiation and performance (rosette production) and % survival of Nanking cherry and Mongolian cherry (Pruski et al., 2005). Treatment Number of rosettes per established culture Mongolian cherry Nanking cherry NAA 0.5 µM + BA 4.4 µM 1.00 (39 1 , 90% 2 ) 0.97 (28, 90%) NAA 0.5 µM + BA 8.9 µM 1.40 (45, 93%) 1.20 (37, 96%) IBA 0.5 µM + BA 4.4 µM 1.30 (30, 96%) 0.93 (27, 80%) IBA 0.5 µM + BA 8.9 µM 1.50 (42, 100%) 1.23 (36, 86%) The results are given as the mean number of rosettes per explant. In brackets are the total number of produced rosettes and the percentage survival; n = 30 per each treatment and species. 1 Total number of rosettes produced out of 30 explants. 2 Survival percentage of cultures. In general, the Nanking cherry cultures are more difficult to establish than the Mongolian cherry. The percentage survival varied from 80% to 96% while the Mongolian cherry was from 90% to 100% being the highest on media recommended above. Rosettes produced on initiation medium (Figure 2B) are placed on solid media in Magenta GA7 vessels for multiplication. In our experiments, shoot proliferation was conducted on MSMO medium with various concentrations of BA and TDZ. Following a number of repetitions we were able to determine optimal concentrations of BA for multiplication of both Mongolian and Nanking cherries. For each species, we calculated the value of BA concentration that maximizes the number of good quality shoots produced by proliferating cultures. Proliferating cultures and their rosettes are shown in Figure 2C. Summary of the regression analysis for shoot proliferation responses as affected by BA concentration is presented in Figure 3. Controls (zero BA) usually produce ca. 1.5–2 shoots, approx. 25 mm long per rosette in both species. With increasing BA concentration in the medium, more but shorter shoots are produced. Above BA concentration of 15.00 µM number of shoots per culture in both species starts to decrease. With increasing BA concentration in the medium length of shoots decreases. Hyperhydricity starts to show on media with high concentrations of BA where cultures become dark green, having short and fragile shoots with poorly developed leaves. In related species, shoot deformations of cultures grown on media with a high content of BA (22.5 µM and above) are known and were reported by Linebeger (1983) for Prunus × Hally Jolivette, Pruski et al. (1990) for Saskatoon berry (Amelanchier alnifolia Nutt.), and Pruski et al. (2000) for chokecherry (P. virginiana L.) and pincherry (P. pensylvanica L.). For both Mongolian and Nanking cherries, BA concentrations 8–15 µM are optimal for
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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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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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