Protocols for Micropropagation of Woody Trees and Fruits

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60 C. HARGREAVES AND M. MENZIES Figure 4. A) Auxin treated shoots set into propagation trays. B) Propagation trays with vented lids in place. C) Rooted tissue-culture shoots 4 weeks from setting. D) Rooted shoots 8 weeks after transfer to Hiko trays. Figure 5. A) Rooted shoots lined out in the nursery bed. B) Tissue-cultured plants after 7 months in the nursery bed and prior to field planting.
ORGANOGENESIS AND CRYOPRESERVATION OF RADIATA PINE and then lined out bare-root, they are transplanted into the nursery bed in late spring/ early summer (November/December), once the roots have bound the container plugs. They are then raised like a seedling or cutting crop, with root conditioning by undercutting, wrenching and lateral root pruning, and may also be topped if plant height exceeds 35–40 cm (Menzies et al., 2005). After one growing season, they will reach a plantable size of at least 30–40 cm (Hargreaves et al., 2005). Growth in the forest. In an early trial planted on five sites in 1983, tissue-cultured plantlets performed as if they were physiologically aged (Menzies et al., 2001), despite being initiated from seed. This was possibly because the organogenesis protocols were still being developed when the plants were produced. Further experience and commercialisation of the tissue-culture systems has improved the protocols, and ensured more uniform and fast early growth (Gleed, 1993). A recent assessment of a field trial established in 1984 (now aged 22 years) has shown that tissue-cultured plantlets were not significantly smaller in diameter than similar genetic quality seedlings and juvenile cuttings (Menzies & Low, unpublished data). Current protocols for organogenesis using axillary shoot multiplication have produced very juvenile plants (Hargreaves et al., 2005), and these are now being evaluated in a field trial. 2.1.7. Storage of in-vitro Cultures (Cryopreservation) Cryopreservation of shoot tip meristems. Successful methods for cryopreservation of radiata pine shoot tip meristems have been difficult to develop. The method presented here was the most successful in a series of 7 preliminary experiments which included a variety of pre-conditioning treatments and regrowth media. The plant material used was a single commercial clone (Christmas Star), there were from 8–65 meristems in each experiment, and all meristems were exposed to liquid nitrogen. The best result was where 12.5% of cryopreserved meristems re-established shoot cultures. This research has subsequently provided a platform for improved results which will be published at a later date (Hargreaves, Towill & Bonnart, 2007). Shoot tip meristems were isolated from in-vitro cultures of Christmas Star (Longview Horticulture, VIC, Australia). Shoots were growing in standard conditions described earlier for shoot multiplication. Meristems were dissected with a stereo microscope, with needles being plucked off until the apical dome of the primary meristem was visible, surrounded by a few needle primordia. Explants were approximately 2 × 1 mm. Isolated shoot tip meristems were collected on dishes of LP medium (no charcoal) and then placed in cryoprotectant preconditioning medium (PGD, modified after Ryynänen, 1996) in 25 ml flasks, at room temperature. Following 2 hours exposure to preconditioning medium, individual meristems were placed into 3 separate drops (5–10 µl) of the PGD solution on 5 × 10 mm strips of aluminium foil. Nunc cryogenic vials were filled with liquid nitrogen and placed in a polystyrene box containing liquid nitrogen. The aluminium foil strips with adhering droplets containing meristems were then plunged vertically into these pre-cooled cryogenic vials. In this manner, meristems were rapidly frozen to –196°C. 61
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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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Page 44 and 45: MICROPROPAGATION OF PINUS PINEA L.
Page 50 and 51: 42 2.1. Organ Culture K. ISHII ET A
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Page 56 and 57: 48 Table 3. Effects of plant growth
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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
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Page 92 and 93: 86 2.1. Explant Preparation M.G. OS
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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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