Protocols for Micropropagation of Woody Trees and Fruits

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MICROPROPAGATION FOR MICROSPORE EMBRYOGENESIS IN OLIVE 369 Figure 4. Sporophytic divisions of olive microspores in culture. Multinucleate microspores with 2(A), 3(B), 4(C), 5(D) nuclei after 15 days of culture. a1–d1: Microspores observed under fluorescence microspore. Nuclei were stained DAPI. a2–1d2: The same microspores under phase contrast.
370 B. PINTOS ET AL. 3. CONCLUSION We present here a new protocol to produce an isolated microspore culture that leads to cell divisions and proembryo formation in Olea europaea L., cv. Arbequina. The morphology of floral buds containing responsive anthers and microspores (open buds with yellow-greenish anthers) from trees grown in the field has also been described. Moreover, also the optimum microspore stage for the induction of pollen division has been determined (from late uninucleate microspore to binucleate pollen stage) as well as the culture conditions for the production of proembryoids from isolated olive microspores have been defined (33°C for 24 h or 3°C during 96 h). In this work, the obtained multicellular olive microspores (proembryos) represent the first step towards haploid olive embryo production. Acknowledgements. This research was supported by project granted by Spanish Ministry of Education and Science (MEC) RTA2005-00118-C02-02 and CAO00-018 from the Consejería de Agricultura y Pesca of the Junta de Andalucía. The collaboration of Drs. J.M. Caballero and C. del Rio, from the CIFA of Córdoba, Spain, is very much appreciated. The authors acknowledge the bilateral agreement between INIA-CSIC Joint Collaboration Program (Convenio CC03-023). Dr. Pintos was recipient of a PhD grant of the National Institute of Agronomic Research (INIA). 4. REFERENCES Barro, F. & Martin, A. (1999) Response of different genotypes of Brassica carinata to microspore culture. Plant Breeding 118, 79–81. Bayliss, K.L., Wroth, J.M. & Cowling, W.A. (2004) Pro-embryos of Lupinus spp. produced from isolated microspore culture. Austr. J. Agricult. Res. 55, 589–593. Bueno, M.A., Gómez, A., Boscaiu, M., Manzanera, J.A. & Vicente, O. (1997) Stress-induced formation of haploid plants through anther culture in cork-oak (Quercus suber L.). Physiol. Plant. 99, 335–341. Bueno, M.A., Gómez, A. & Manzanera, J.A. (2000) Somatic and gametic Embryogenesis in Quercus suber L. In Jain, S.M., Gupta, P.K. & Newton, R.J. (Eds). Somatic Embryogenesis in Woody Plants. Forestry Sciences. Vol 6. Kluwer Academic Publishers. Dordrecht/Boston/London. ISBN 0-7923- 6419-8, pp. 479–508. Bueno, M.A. & Manzanera, J.A. (2003) Oak anther culture. In Doubled Haploid Production in Crop Plants. A Manual. Maluszynski, M., Kasha, K.J., Foster, B.P. & Szarejko, I. (Eds). Kluwer Academic Publishers, Dordrecht. ISBN 1-4020-1544-5, pp. 297–301. Bueno, M.A., Pintos, B., Gomez, A. & Martin, A. (2004) Monitoring of in vitro cultured microspores switched to embryogenesis in Olea europaea L. Polen: Proc. XI International Palynological Congress, 14, pp. 26. Bueno, M.A., Pintos, B., Höfer, M. & Martin, A. (2005) Pro-embryos induction from Olea europaea L. isolated microspore culture. Acta Physiol. Plant. 27, 695–701. Chupeau, Y., Caboche, M. & Henry, Y. (1998) Androgenesis and haploid plants., Springer-Verlag, Berlin, Heidelberg. Germana, M.A. & Chiancone, B. (2003) Improvement of the anther culture protocol in Citrus clementina Hort. ex Tan. Plant Cell Rep. 22, 181–187. Höfer, M. (2004) In vitro androgenesis in apple – improvement of the induction phase. Plant Cell Rep. 22, 365–370. Höfer, M., Touraev, A. & Heberle-Bors, E. (1999) Induction of embryogenesis from isolated apple microspores. Plant Cell Rep. 18, 1012–1017. Kyo, M. & Harada, H. (1986) Control of the developmental pathway of tobacco pollen in vitro. Planta 168, 427–432.
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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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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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