Protocols for Micropropagation of Woody Trees and Fruits

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MICROGRAFTING OF PISTACHIO Constituent mg/L Constituent mg/L Basal Salts Ammonium nitrate 1650.0 Boric acid 6.20 Calcium chloride anhydrous 332.20 Cobalt chloride hexahydrate 0.0250 Cupric sulphate pentahydrate 0.0250 Disodium EDTA dehydrate 37.260 Ferrous sulphate heptahydrate 27.80 Magnesium sulphate anhydrous 180.70 Manganese sulphate monohydrate 16.90 Potassium iodide 0.830 Potassium nitrate 1900.0 Potassium phosphate monobasic 170.0 Sodium molybdate dehydrate 0.250 Zinc sulphate heptahydrate 8.60 Organic Additives Organic Additives 291 Sucrose Varies Casein hydrolysate Varies Varies Myo-Inositol 100.0 Thiamine hydrochloride 10.0 Nicotinic acid (Free acid) 1.0 Pyridoxine hydrochloride 1.0 Agar Varies pH 5.7 RPM SPM CMM MDM Rootstock Production Medium Scion Production Medium 1 Culture Maintenance Medium Micrograft Development Medium Sucrose 20.000 30.000 30.000 30.000 N 6 -Benzyladenine 0.5 1.0 0.5 0.5 Casein hydrolysate – 200 200 – l-ascorbic acid – 200 200 200 Agar 5700 5700 5700 7000 1 Table 1. Pistachio basic culture media (BCM). L-ascorbic acid Table 2. Formulations of pistachio micrografting media . All units are in mg/L (or ppm). All chemical used were obtained from Sigma Ltd. The pH of all media is adjusted to 5.7.
292 A. ONAY ET AL. Levels of survival, the average shoot length and the average shoot number were assessed after 5 weeks of culture. The position of the explants cultured did not influence the survival rate but the highest length of shoots was obtained with the apical tips explanted from the pruned shoot trunks. Explants from the pruned shoot trunk produced most shoots and means were greater than the explants excised from other positions. 1. Collect the actively growing shoot tips (3–5 cm long) from a pruned trunk of mature pistachio by cutting with a sharp scalpel and cover immediately the cut edge with a moistened cotton material. 2. Bring the shoot tips to the laboratory, chop off the leaves (with petioles) and surface sterilize or dip the defoliated shoot tips into the sterile distilled water (SDW) if they were not used for initiation of cultures. Explant sterilization and initiation of cultures. In fact, it is difficult to establish in vitro cultures from lignified stem sections from mature pistachio trees during the dormant season. We have been conducting research on forcing shoots for years. Shoots were forced in the greenhouse under a 24 h photoperiod until they were large enough to excise, disinfest surface and place in vitro. 1. Remove the stem section of the explants (use only shoot tips, 15–20 mm long) and sterilize them. 2. Treat with 20% commercial bleach with 14% available chlorine for 35 min on a shaker at 150 rpm. 3. Gently wash with SDW at least 3 times for 5 min. 4. Cut the basal end of the rinsed explants. 5. Shake the explants with SDW at least twice for 1 h on horizontal type shaker at 150 rpm with SDW. 6. Inoculate 3–5 mm apical tips on scion production medium (SPM) (Table 2). 7. Incubate cultures in a growth room under the light (at 40 µmol m –2 s –1 photo- synthetic photon flux density; fluorescent lamps, 75W) and a photoperiod of 16 h at 25 ± 2°C for 4 weeks. 8. Excise the sprouting apical tips with minimal mother tissue attached. Note that shoot tip region consist of apical meristematic bud surrounded by a number of leaf primordia encased by fleshy green-white tissue of a leaf sheath. 9. Transfer the regenerated shoots to the shoot culture maintenance medium (CMM) (Table 2). 10. Incubate until the explants are at 3–4 leaf stage. An early subculture is recommended if the apical tips start releasing the polyphenols in the culture medium. 11. Use freshly sprouted apical tip or healthy growing shoot tips (4–6 mm long) as scions (Figure 1A).
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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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Page 252 and 253: MICROGRAFTING IN GRAPEVINE 253 Tabl
Page 254 and 255: 2.4. Culture Conditions MICROGRAFTI
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Page 266 and 267: CHAPTER 25 APRICOT MICROPROPAGATION
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Page 274 and 275: 2.4. Acclimatization APRICOT MICROP
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Page 288 and 289: CHAPTER 27 MICROGRAFTING OF PISTACH
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Page 322 and 323: CHAPTER 30 IN VITRO MUTAGENESIS AND
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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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