Protocols for Micropropagation of Woody Trees and Fruits

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42 2.1. Organ Culture K. ISHII ET AL. 2. EXPERIMENTAL PROTOCOL 2.1.1. Materials 1. Mature seeds collected from late August to early September from remaining trees (Figure 1A) of Pinus armandii var. amamiana in Yakushima island. 2. Laminar-flow chamber, Petri dishes, forceps, scalpel, pipettes. 3. Ethanol, sterile distilled water, culture tubes, culture flasks. 4. Dissecting microscope. 5. Media (see Table 1). 2.1.2. Methods The regeneration method can be divided into three main steps: initial culture, shoot elongation, and rooting. Initial culture. For elimination of empty seeds caused by inbreeding depression, only submerged seeds in 100% ethanol were used for further experiments in mature seeds. 1. Remove the seeds from the cones. 2. Sterilize the seeds with 70% ethanol for 3 min. 3. Wash the seeds two times with distilled water. 4. Mature embryos were excised from the seeds. 5. Transfer the sterile embryos on to the induction medium in the test tubes. Mainly a half strength DCR (Gupta & Durzan, 1985) medium with different concentration of plant growth regulators (2 or 10 µM BAP plus 0.1 µM NAA, 0.4, 2 and 10 µM BAP) was used. Culture tubes (18 mm i.d. × 160 mm) containing 15 ml of agar solidified media were used for initial culture, and 200 ml culture flasks containing 70 ml agar-solidified medium were used for subculture. Shoot elongation 1. Regenerated buds with embryos are transferred to shoot elongation medium containing 2g/l activated charcoal (Figure 1B) (Table 1). 2. Elongated shoots are divided and further subcultured on the same medium for 1–2 months interval. Rooting 1. Shoots more than 1.5 cm length are cut and transferred to the rooting medium. 2. For rooting of shoots, RIM medium (Abo El-Nil & Milton, 1982) containning indole butyric acid (IBA) was used. 3. Rooted shoots (Figure 1C) are subcultured to the hormone free florialite (Nisshinbo, Japan) medium containing 0.1% hyponex . ® ®
MICROPROPAGATION OF PINUS ARMANDII VAR. AMAMIANA Table 1. Media for organ culture of Pinus armandii var. amamiana. Chemicals Initial culture Shoot elongation Rooting (mg/l) (1/2 DCR) (1/2 DCR + AC) (RIM) NH4NO3 200 200 KNO3 170 170 187.5 Ca(NO3)2 × 4H2O 152 MgSO4 × 7H2O 185 185 200 CaCl2 × 2H2O 42.5 42.5 NaH2PO4 × H2O 138 KH2PO4 85 85 170 K2SO4 860 Na2SO4 200 KCl 65 FeSO4 × 7H2O 13.9 13.9 27.8 Na2EDTA 18.65 18.65 37.3 MnSO4 × 4H2O 11.15 11.15 5 ZnSO4 × 7H2O 4.3 4.3 0.5 H3BO3 3.1 3.1 10 KI 0.415 0.415 1 Na2MoO4 × 2H2O 0.125 0.125 0.1 CuSO4 × 5H2O 0.0125 0.0125 0.1 CoCl2 × 6H2O 0.0125 0.0125 0.1 NiCl2 0.0125 0.0125 Myo-inositol 100 100 100 Nicotinic acid 0.5 0.5 5 Pyridoxine HCl 0.1 0.1 0.5 Thiamine HCl 1 1 5 Glutamine 100 Glycine 1 1 Coumarin 0.0146 Riboflavin 0.2 BAP 0.045–2.25 NAA 0–0.0186 IBA 1–3 Activated charcoal 2000 Sucrose 15000 15000 15000 Agar 8000 9000 8000 43
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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.
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Page 54 and 55: 46 Chemicals (mg/l) K. ISHII ET AL.
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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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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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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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