Protocols for Micropropagation of Woody Trees and Fruits

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378 L. TIAN AND S.I. SIBBALD 2.4. Rooting Transfer individual shoots to rooting medium (Table 1) in a 100 × 25 mm petri dish for rooting. Rooting efficiency varies with each plum variety. Use of NAA can increase the rooting efficiency, especially for the varieties with poor rooting efficiency (Tian et al., 2007a). Roots generally appear after 4 to 8 weeks (Figure 2F). Each shoot produces 1 to 2 main roots with a few small rootlets. Carefully move rooted plantlets to 10 cm Magenta containers (Magenta Corp., Chicago) containing the same rooting media for plant development (Figure 2G). Roots are prone to breaking off at this point if care is not taken to first loosen the media around them. 2.5. Hardening When plantlets have reached 3–5 cm in height, gently remove them from the Magenta containers and remove any agar from the roots. Transfer the plantlets to 12cm pots containing moistened commercially available potting mix. Keep soil moist and the plantlets covered with empty Magenta containers or transparent plastic bags during the first week. After this period of acclimation, remove the Magenta containers and expose the plantlets to standard greenhouse conditions. Plants usually establish in the greenhouse in about 2 weeks. Fertilize weekly with N-P-K fertilizer (20-20-20). 2.6. Field Testing Plants that are well established in the greenhouse (Figure 2H) and with a size of about 1 meter can be transferred to the field. 2.7. Storage of In Vitro Cultures In vitro cultures can be maintained for months or years as shoot cultures. Each month, divide multiple shoots into single shoots and transfer to fresh induction medium (Figure 2D) in order to keep the cultures in a healthy condition. Over the month, these shoots will produce multiple shoots and the process can be repeated. In this fashion, cultures have been maintained for several years, with up to 98% of the shoots continuing to multiply after one year. 3. CONCLUSION Although different in taxonomy and other aspects, Japanese plum and European plum can be regenerated via epicotyls and/or hypocotyls using a similar approach. Mature seeds are used as the source of explants and thus the research materials can be supplied year around. The plant recovery time is relatively quick. It usually takes about 4 to 5 months from the introduction of the explant into culture to plant establishment in the greenhouse. Plants can be regenerated from seeds of different plum varieties. New shoot development from induced primary shoots is a useful way for maintaining the culture in the laboratory. A large number of plants can be produced via culture of epicotyls or hypocotyls, or via secondary shoot formation from the developed shoots.
MICROPROPAGATION OF PLUM USING MATURE SEEDS 379 Acknowledgements. We would like to thank Kristine Leach and Yan Wen for assistance in the development of this protocol. 4. REFERENCES Bellini, E., Nencetti, V. & Nin, S. (2002) Genetic improvement of plum in Florence. Acta Hort. 577, 19–24. Capote, N., Gorris, M.T., Martinez, M.C., Sensio, M., Olmos, A. & Cambra, M. (2006) Interference between D and M types of plum pox virus in Japanese plum assessed by specific monoclonal antibodies and quantitative real-time reverse transcription-polymerase chain reaction. Phytopathology 96, 320–325. Gamborg, O.L., Miller, R.A. & Ojim, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50, 151–158. Gonzalez Padilla, I.M., Webb, K. & Scorza, R. (2003) Early antibiotic selection and efficient rooting and acclimatization improve the production of transgenic plum plants (Prunus domestica L.). Plant Cell Rep. 22, 38–45. Kaufmane, E., Ikase, L., Trajkovski, V. & Lacis, G. (2002) Evaluation and characterization of plum genetic resources in Sweden and Latvia. Acta Hort. 577, 207–213. Lloyd, G. & McCown, B. (1981) Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot tip culture. Int. Plat. Prop. Soc. Proc. 30, 421–427. Mante, S., Morgens, P.H., Scorza, R., Cordts, J.M. & Callahan, A.M. (1991) Agrobacterium-mediated transformation of plum (Prunus domestica L.) hypocotyl slices and regeneration of transgenic plants. Bio/Technology 9, 853–857. Murashige, T. & Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 473–497. Okie, W.R. & Ramming, D.W. (1999) Plum breeding worldwide. HortTechnology 9, 162–176. Scorza, R., Hammerschlag, F.A. & Zimmerman, T.W. (1995) Genetic transformation in Prunus persica (peach) and Prunus domestica (plum). In Bajaj, Y.P.S. (Ed) Biotechnology in Agriculture and Forestry. 34: Plant Protoplasts and Genetic Engineering VI, Springer-Verlag, Heidelberg pp. 255–268 Sibbald, S., Wang, X., Wen, Y. & Tian, L. (2006) Genetic transformation of prunus domestica using hygromycin resistance as selection for plum pox virus research. Joint Meeting of the American and Canadian Phytopathological Societies & Mycological Society of America. July 29–Aug 2, 2006. Quebec City, Quebec, Canada. Szabo, Z. & Nyeki, J. (2002) Japanese plum in Hungary. Acta Hort. 577, 65–71. Tian, L., Brown, D. & Sibbald, S. (2004) Regeneration and genetic transformation of Prunus domestica grown in canada for plum pox virus resistance. World Congress of In Vitro Biology. May 22–26, 2004. San Francisco, California, USA. Tian, L., Wen, Y. & Sibbald, S. (2005) Regeneration of Prunus salicina from embryonic axes. 2005 In Vitro Biology Meeting. June 5–8, 2005. Baltimore, MD, USA. Tian, L., Sibbald, S., Subramanian, J. & Svircev, A. (2007a) Characterization of Prunus domestica L. in vitro regeneration via hypocotyls. Sci. Hort. 112, 462–466. Tian L, Wen, Y., Sibbald, S. & Subramanian, J. (2007b) Regeneration of Prunus salicina Lindl from hypocotyls of mature seeds. Revised and submitted.
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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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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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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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