Protocols for Micropropagation of Woody Trees and Fruits

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APRICOT MICROPROPAGATION 2.2.2. Hyperhydricity Hyperhydricity is a physiological disorder frequently related to the in vitro environment during micropropagation of woody plants. Apricot shoot tips at the prolife- ration stage are very sensitive to hyperhydration (Figure 5), although the degree of symptoms is genotype-dependent. Treatments that decrease hyperhydricity, without affecting micropropagation rates in apricot, are the application of a bottom cooling system for 1 or 2 weeks in each culture cycle; and an increase in the agar concentration of the culture medium (0.6–0.8%) or the use of Agargel TM (0.5%) as gelling agent (Pérez-Tornero et al., 2001). However, the effectiveness of each method depends on the genotype and only bottom cooling have consistently prevented hyperhydricity in all apricot cultivars tested. Figure 5. Hyperhydric apricot explants of two different cultivars. Bottom cooling system. High relative humidity (RH) is probably the most important environmental factor that induces hyperhydricity. The bottom cooling system (Figure 6) decreases the RH inside the in vitro culture jar by condensing water on the cooled culture medium (Vanderschaeghe & Debergh, 1987). Vessels are placed on an iron plate cooled by a system of copper tubes. The water, cooled by a cryostat (Selecta, Frigiterm-10), circulates constantly through the tubes. The temperature of the water is adjusted to 13° C in the cryostat to obtain a gradient of approximately 4° C between the bottom and the top of the jar. RH within the jar is then decreased to approximately 75% (Vanderschaeghe & Debergh, 1987). Hyperhydricity can be avoided by placing proliferating shoots in the bottom cooling system in the first week of the culturing cycle. More than 64% of hyperhydric shoots could be returned to normal by keeping them for 3 weeks in the bottom cooling system. 273
274 2.3. Rooting O. PÉREZ-TORNERO AND L. BURGOS 2.3.1. Culture Medium and Conditions QL macronutrients diluted to half strength, DKW micronutrients, organics and vitamins (Table 1), plus 40 mg/l phloroglucinol, are used for the rooting of apricot shoots. Sucrose is used at 2%, agar at 0.6% (Hispanlab, S.A.) and IBA at 1 to 3 µM, depending on genotype (Pérez-Tornero & Burgos, 2000). Figure 6. Bottom cooling system. Medium is prepared, dispensed and sterilized as described previously. Culture conditions are as described above. Proliferating shoots should be at least 2 cm long. The basal callus is eliminated and shoots are cultured for 21 days in the rooting medium, where induced adventitious roots grow (Figure 7). Although rooting of micropropagated apricot shoots is not a problem, a high percentage of shoots shows symptoms of apical necrosis on the rooting media, sometimes exceeding 80% and also affecting axillary buds. Plantlets with necrotic apices frequently are unable to grow and cannot be acclimatized. This effect is highly genotype-dependent. 2.3.2. Apical Necrosis during Rooting Apical necrosis can be related to lower endogenous cytokine concentration in the shoot apex (Kataeva et al., 1991) in apricot (Pérez-Tornero & Burgos, 2000). Apical necrosis during rooting can be solved by immersing shoot tips in concentrated BA solution (22.19 to 44.38 µM) just before explanting shoots on the rooting media. It is necessary to dry the excessive BA solution to avoid any interference with rooting.
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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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Page 248 and 249: CHAPTER 23 MICROGRAFTING IN GRAPEVI
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Page 252 and 253: MICROGRAFTING IN GRAPEVINE 253 Tabl
Page 254 and 255: 2.4. Culture Conditions MICROGRAFTI
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Page 258 and 259: CHAPTER 24 MICROGRAFTING GRAPEVINE
Page 260 and 261: MICROGRAFTING GRAPEVINE FOR VIRUS I
Page 266 and 267: CHAPTER 25 APRICOT MICROPROPAGATION
Page 268 and 269: APRICOT MICROPROPAGATION Figure 1.
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Page 278 and 279: CHAPTER 26 IN VITRO CONSERVATION AN
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Page 288 and 289: CHAPTER 27 MICROGRAFTING OF PISTACH
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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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