Protocols for Micropropagation of Woody Trees and Fruits

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CHAPTER 13 PROPAGATION OF SELECTED PINUS GENOTYPES REGARDLESS OF AGE R. RODRÍGUEZ 1,2 , , P. SÁNCHEZ 1 , M.F. FRAGA 3 , M. BERDASCO 3 , R. HASBÚN 1,2,6 , J.L. RODRÍGUEZ 1,2 , J.C. PACHECO 4 , I. GARCÍA 5 , M.M. URIBE 6 , D. RÍOS 6 , M. SÁNCHEZ-OLATE 6 , M.E. MATERÁN 7 , C. WALTER 8 AND M.J. CAÑAL 1,2 L. VALLEDOR 1,2 1 Lab. Fisiología Vegetal, Dpto. Biología de Organismos y Sistemas, Universidad de Oviedo. c/ Catedrático Rodrigo Uria s/n 33071 Oviedo, Spain. E-mail: 2 3 National Cancer Center (CNIO). Melchor Fernández Almagro, 3. Madrid, Spain. 4 Universidad tecnológica y pedagógica de Colombia, Carretera Central del Norte, Tunja, Boyacá, Colombia. 5 Instituto Viveros Oihamberri S.A., c/ Olabarrita Berri s/n 48111 Laukiz, Vizcaya, Spain. 6 Facultad de Ciencias Forestales, Dpto. Silvicultura- Centro de Biotecnología, Universidad de Concepción. Casilla 160-C, Concepción, Chile. 7 mjccanal@uniovi.es Instituto de Biotecnología de Asturias, Oviedo, Spain. Spanish Centro Biotecnológico de Guayana, Universidad Nacional Experimental de 8 Guayana. Av. Atlántico Puerto Ordaz, Estado Bolívar, Venezuela. Forest Research Institute Ltd. Sala Street, Private Bag 3020, Rotorua, New Zealand 1. INTRODUCTION Most of pine trees do not reproduce naturally from sprouts. Reproduction from artificially rooted propagules of pines has been proved as a successful reproduction method in several countries. This leads us to two different ways to obtain artificial reproduction. Hedging method is used for mass-producing large number of symmetrical and straight cuttings and for maintaining the juvenile nature of propagules (Libby et al., 1972). It has been widely used in breeding programs of plant species for preservation and multiplication of the desired genotypes to establish seed orchards. In this technique we include micrografting which is useful to rejuvenate adult material (Fraga et al., 2002b). Plantlet regeneration from embryos and cotyledons is another approach (Aitken-Christie et al., 1981), it has a great potential for forest tree 137 S.M. Jain and H. Häggman (eds.), Protocols for Micropropagation of Woody Trees and Fruits, 137–146. © 2007 Springer.
138 R. RODRÍGUEZ ET AL. multiplication compared to other alternative techniques, such as cutting propagation. In this chapter we describe protocols for in vitro micropropagation of selected Pinus species like radiata pine (Pinus radiata D. Don), austrian pine (Pinus nigra Arn.) and caribbean pine (Pinus caribaea Mor). 2. IN VITRO APPROACHES Micropropagation is an attractive alternative for the multiplication of selected genotypes. Taking into account the actual barrier that exists in this area for achieving industrial plant production in the following sections we will pay special attention to micropropagation techniques, considering the available information from previous reports and also our own experience with juvenile and mature radiata, austrian and caribbean pines (Rodríguez, 1990; Fraga et al., 2002a,b, 2003; Uribe et al., 2004; Rodríguez et al., 2005b). A successful micropropagation technique is strongly influenced by the genotype and chronological/physiological age of the donor plant (Rodríguez et al., 2004a,b,c, 2005b). This work is presented in three sections: I) Morphogenesis from embryos and seedlings (Figures 1 and 2a), II) Micropropagation from trees younger than 5 years (Figure 2b), and III) Micropropagation from mature trees (Figure 2c–e). Thereby, the extent of currently available information (Horgan, 1987; Smith, 1997; Fraga et al., 2002a,b; Diego et al., 2004; Rodríguez et al., 2004c) would be of great help to overcome the limitations facing in vitro plant propagation. 2.1. Explants and Sterilization Explant selection depends on the final aim of the program. The selection of different explants has shown significant influence on shoot production (Aitken-Christie et al., 1981). Embryos and cotyledons. To proceed, imbibe seeds in running tap water for 24 h and then stratify in plastic bags for 2 days at 4 C. Seeds are surface sterilized in 5% (v/v) sodium hypochlorite and then washed in sterile water. For embryo culture, dissect embryos from seeds under sterile conditions, or excise cotyledons after aseptically germination (Villalobos & Engelmann, 1995). There is another similar protocol, place seeds for 30 min in 3% sodium hypochlorite solution containing 0.05% (w/v) of Tween ® ° 20 followed by five rinses with sterile water. Once the seed coat is removed the megagametophyte is re-sterilized for 15 min in 15% bleach and for 5 min in hydrogen peroxide (Muriithi et al., 1993). We have proved that mild treatments, 7.5% sodium hypochlorite (v/v) 45 min or ethanol 50% (v/v) 90 min, followed by three rinses in sterile water and after removing seed coats, also work efficiently. Mature tissues. This type of tissues is difficult to sterilize, and propagation of adult trees can be initiated with different tissues. Apical buds or healthy branch tips are usually collected from vigorous trees, but best responses are obtained when caulinar
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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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Page 152 and 153: ROOT INDUCTION OF PINUS SYLVESTRIS
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Page 156 and 157: CHAPTER 15 MICROPROPAGATION OF BETU
Page 158 and 159: MICROPROPAGATION OF BETULA PENDULA
Page 166 and 167: CHAPTER 16 PROTOCOL FOR DOUBLED-HAP
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Page 182 and 183: CHAPTER 17 IN VITRO PROPAGATION OF
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Page 188 and 189: Axillary shoots (number) IN VITRO P
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IN VITRO PROPAGATION OF FRAXINUS SP
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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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