Protocols for Micropropagation of Woody Trees and Fruits

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DOUBLED-HAPLOID MICROPROPAGATION IN QUERCUS SUBER L. Figure 5. Scheme for the maturation of cork oak doubled-haploid embryos. A) Spontaneously induced doubled-haploid embryos. B) Doubled-haploid embryos induced by anti-mitotic agents. C) Doubled-haploid embryos on maturation medium. D) Cork oak doubled-haploid embryos alter one month on maturation medium. 2.5. Maturation of Doubled-haploid Embryos Both the anther embryos that spontaneously duplicated their genome and those diploidized by the action of oryzalin were subjected to treatments inducing their maturation (Figure 5A,B). The addition of 1% activated charcoal to the maturation medium provoked a significant increment of both size and weight of the doubledhaploid embryos (Figure 5C,D). Immature translucent doubled-haploid embryos from either origin were cultured at the cotyledonar stage, about 3–4 mm size and 30 mg fresh weight, on SM basal 171
172 B. PINTOS ET AL. medium supplemented with 3% (w/v) sucrose, 1% activated charcoal and solidified with 0.8% (w/v) agar, pH = 5.6. Then the cultures were subjected to a temperature of 25±1° C in darkness for one month and next at 4° C during two months (Pintos, 2005). 2.6. Germination of Doubled-haploid Embryos Mature cork oak doubled-haploid embryos (Figure 6A) were prepared for the germination treatments by immersion in sterile distilled water for 24 h at 4° C. The best germination rates (18.7%) were obtained when the mature embryos were cultured on basal SM medium supplemented with 10 g/L agar, 15 g/L sucrose, 0.1 mg/L IBA and 0.05 mg/L BAP. (Figure 6B). The pH of the medium was adjusted to 5.7 and the embryos were cultured under a photoperiod of 16 h light and 8 h darkness. Temperature: 25 ± 1° C. 2.7. Acclimation of Doubled-haploid Embryos Plantlets germinated in the previous medium were carefully taken out of the test tube and agar was washed from the roots with tap water. Those plantlets were then transferred to nursery pots containing a mixture of peat:perlite:vermiculite 1:1:1 (Figure 7). Afterwards, the plantlets were subjected to a preventive fungicide treatment with 1.8 g/L PREVICUR ® (propamocarb). The first days, plantlets were acclimated in a chamber with high relative moisture, close to 100% (Figure 8). After acclimation, those plantlets were transferred to bigger pots to permit further growth. Three growth seasons later, they were transferred to soil (Bueno & Manzanera, 2003). A total of fourteen doubled-haploid cork oak plantlets from anther culture were acclimated. 2.8. Marker-assisted Verification A problem found in the induction of embryogenesis through anther culture was the actual origin of those embryos. Totipotency of plant cells would permit the regeneration either from haploid cells of the anther cavity (microspores) or from diploid cells of the anther wall. The genetic composition of both types of regenerants would differ. Thanks to flow cytometry, we observed in cork oak that the main composition of the regenerated embryos was haploid (89.03%), which clearly indicated their haploid origin, in this case microspores. Nevertheless a small percentage were either diploid (7.87%) or triploid (1.24%) (section 2.3). In order to elucidate if those embryos were originally haploids which spontaneously duplicated or triplicated their genome, or alternatively those embryos were regenerated from the diploid tissue of the anther wall, a genetic test was designed. Analyses were performed through microsatellite markers.
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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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Page 128 and 129: CHAPTER 12 MICROPROPAGATION OF LARI
Page 130 and 131: MICROPROPAGATION OF LARIX SPECIES V
Page 140 and 141: CHAPTER 13 PROPAGATION OF SELECTED
Page 142 and 143: PROPAGATION OF SELECTED PINUS GENOT
Page 150 and 151: CHAPTER 14 ROOT INDUCTION OF PINUS
Page 152 and 153: ROOT INDUCTION OF PINUS SYLVESTRIS
Page 154 and 155: ROOT INDUCTION OF PINUS SYLVESTRIS
Page 156 and 157: CHAPTER 15 MICROPROPAGATION OF BETU
Page 158 and 159: MICROPROPAGATION OF BETULA PENDULA
Page 166 and 167: CHAPTER 16 PROTOCOL FOR DOUBLED-HAP
Page 168 and 169: DOUBLED-HAPLOID MICROPROPAGATION IN
Page 182 and 183: CHAPTER 17 IN VITRO PROPAGATION OF
Page 184 and 185: IN VITRO PROPAGATION OF FRAXINUS SP
Page 188 and 189: Axillary shoots (number) IN VITRO P
Page 196 and 197: CHAPTER 18 MICROPROPAGATION OF BLAC
Page 198 and 199: MICROPROPAGATION OF BLACK LOCUST 19
Page 200 and 201: 2.5. Rooting and Acclimatization MI
Page 202 and 203: MICROPROPAGATION OF BLACK LOCUST 19
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Page 216 and 217: MICROPROPAGATION OF SALIX CAPREA L.
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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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