Protocols for Micropropagation of Woody Trees and Fruits

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MICROPROPAGATION OF MEDITERRANEAN CYPRESS exposure of explants to cytokinins (Capuana & Giannini, 1997). Recovery of juvenile traits can be induced through rejuvenating treatments, such as repeated grafting, etiolation and hedging (Fouret et al., 1985; Huang et al., 1992; Giovannelli & Giannini, 2000). However less than 20% of these “re-invigorated” shoots are able to elongate up to 15 mm in 4 weeks on EM. The number of elongated shoots per explant is not improved by maintaining the explants on EM for 6 or 8 weeks; however, the medium is able to increase the length of the longer shoots, i.e., a high number of very long shoots (over 25 mm) has been obtained from each culture vessel. Besides, dividing the elongated shoots into segments (from 5 to 10 mm) has a negative effect on successive growth. As for juvenile explants, shoots longer than 15 mm are then excised from primary explants and cultured again on PM. Under these conditions, juvenile traits disappear slowly and after five proliferation cycles, the oldest material shows a significant decrease of the proliferation capacity (from 3.5 to 2 mean number of axillary shoot per explant) and an opposite decussate phyllotaxy is newly reached (leaf adult trait). However, significant clonal differences in morphogenetic capacity have been found in in vitro competence and persistency of juvenile traits. For instance, primary explants of clones 329 and 771 show a reduced reactivity in vitro (lower than 5%) and axillary shoots fail to elongate even after repeated subcultures on EM. In our laboratory, stabilised cultures have been obtained only in the case of five out of 16 clones and it has been maintained for more than five proliferation cycles but in vitro growth performances of these clones have remained lower than juvenile explants (unpublished data). For root induction, shoots longer than 20 mm with juvenile morphological traits and derived from stabilized cultures of five grafted clones and one 150-year-old plant, are rooted following the procedure utilised for juvenile material. After 8 weeks on solid medium only two grafted clones and the 150-year-old plant are able to form adventitious roots but at low rates (from 6 to 15%). Besides, a high percentage (>60%) of shoot tip necrosis has been recorded in all clones. Taken together, these results show that the appearance of juvenile leaf traits did not support the hypothesis that a rejuvenation process will take place during in vitro culture. Culture conditions. During shoot proliferation and elongation, cultures are incubated at 25 ± 1°C under a 16-h photoperiod at an irradiance of 80 µE m –2 s –1 . 2.2.3. Adventitious Bud Formation In vitro establishment and adventitious bud induction. Adventitious buds are formed on excised mature embryos cultured on BIM (Bud induction medium) with 10 µM BA for 10 days. After induction, embryos with adventitious buds are transferred to the same medium without hormones to induce bud development. After 3–4 weeks, an average of 20 adventitious buds per embryo are recorded and after 24 weeks, it is possible to obtain from 2 to 10 adventitious shoots longer than 10 mm (Lambardi et al., 1995). The adventitious shoots, separated from primary explants, are cultured following the same protocol used for axillary shoot. 99
100 A. GIOVANELLI AND A. DE CARLO Figure 1. Micropropagation of C. sempervirens. A) Axillary budding (arrows) in adult explant (bar = 2 mm). B) 500-ml glass jar with microshoots on proliferation medium (PM) (bar = 17 mm). C) Rooted explant after 8 weeks on solified expression medium (bar = 10 mm). D) Embryogenic tissue (bar = 1 mm). E) Embryonal and suspensor region (bar = 0.3 mm). F) Cluster with multiple elongating shoots obtained by organogenesis from callus after 6 months in culture (bar = 5 mm).
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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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Page 64 and 65: 56 2.1.3. Organogenesis from Field-
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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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Page 98 and 99: CHAPTER 9 MICROPROPAGATION OF MEDIT
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Page 102 and 103: MICROPROPAGATION OF MEDITERRANEAN C
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Page 126 and 127: 122 D. EWALD Root development. Afte
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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
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Page 152 and 153: 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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Protocols for Micropropagation of Woody Trees and Fruits

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