Protocols for Micropropagation of Woody Trees and Fruits

More documents

Recommendations

Info

3500 3000 2500 2000 1500 1000 500 0 1000 500 0 1500 1000 500 0 500 0 PLANT GENETIC FIDELITY 79 150 180 210 240 270 300 330 360 390 420 ssrQpZAG36 (FAM) ssrQpZAG36 (FAM) ssrQpZAG36 (FAM) ssrQpZAG36 (FAM) Donor tree Undifferentiated callus Somatic embryo (SE2) Converted plant Figure 2. Amplification of the dinucleotide ssrQpZAG36 (FAM) locus in Quercus suber (from top to bottom): donor tree, an undifferentiated callus, a normal dicotyledonary somatic embryo (SE2) and a converted plant. All electropherograms show heterozygous individuals with two alleles of approximately 209 and 219 bp. Top scale indicates fragment size in nucleotides. Left scale indicates fluorescence intensity measured in relative fluorescence units (adapted from Lopes et al., 2006). Microsatellite markers/loci that can be amplified with minimum non-specific annealing, and that even have overlapping size ranges, can be separated efficiently and simultaneously in automated sequencers as there are dyes that fluoresce at different wavelengths (Karp et al., 1997). In this case, several PCR products can be pooled
80 C. SANTOS ET AL. (i.e. multiple loading/multiloading). Alternatively, several loci/markers can be coamplified during PCR (i.e. multiplexing); this can be particularly useful when a great number of loci are to be amplified and/or when limited amounts of DNA are available. Optimizing a multiplex PCR system may present many difficulties, although there are now some multiplex PCR commercial kits available. For oak species PCR multiplexing procedures are available in the literature. 2.2.3. Troubleshooting 1. Although rare with this protocol, the problem of disappearing microsatellites, whereby previously scorable samples fail to appear when a PCR is repeated, can sometimes be avoided by pipetting the master mix onto each sample individually and ensuring it is mixed with the template DNA. 2. When analysing the fragment size in the automated sequencer, the fluorescence intensity of a specific sample may change dramatically when amplifying different microsatellite markers (using different fluoro-labels). Therefore, after PCR it is advisable to run samples on an agarose gel to try and quantify the PCR product. Alternatively, at first it is better to run only a few samples on the automated sequencer to estimate whether scorable readings are obtained or not. If fluorescence is too intense that it prevents the obtaining of a result, a water-dilute must be done to the PCR sample. Conversely, if fluorescence intensity is too low, it is very likely that the PCR was not successful. In this case a repetition of the PCR run and/or optimization of PCR conditions are needed. 3. Many microsatellites show, besides stutter bands, an additional band above the expected allele size. This is due to the activity of the terminal transferase of Taq DNA polymerase which adds an adenine to the PCR product (the plus-A phenomenon described by Clark, 1988). Note that this terminal transferase activity is polymerase- and PCR primer-dependent and if it cannot be surpassed by PCR optimization, the microsatellite in question must be discarded. 3. CONCLUSION Once a micropropagation protocol is well established and performed routinely (as is the case of Quercus suber somatic embryogenesis) it is of utmost importance to verify the fidelity of the in vitro derived material, particularly, when the objective is to use the genetic material in a plant breeding programme. Due to the existing constraints of the current genetic screening tools and to the different types of genetic modifications, the combined use of different techniques able to provide complementary data is recommended. In the present chapter, detailed protocols for the application of the highly sensitive and reproducible techniques of flow cytometry and nuclear microsatellites are given. Also, some best practices concerning plant fidelity using the FCM and microsatellite analysis in Q. suber are highlighted here, and can be transposed to regeneration processes of other species. Both techniques can be used at different stages of any
Page 2 and 3:
PROTOCOLS FOR MICROPROPAGATION OF W
Page 4 and 5:
A C.I.P. Catalogue record for this
Page 6 and 7:
vi TABLE OF CONTENTS 14. Root induc
Page 8 and 9:
viii TABLE OF CONTENTS 43. Micropro
Page 10 and 11:
x PREFACE on precise stepwise proto
Page 12 and 13:
CHAPTER 1 TOTIPOTENCY AND THE CELL
Page 14 and 15:
TOTIPOTENCY AND THE CELL CYCLE 5 a
Page 16 and 17:
2.2. Plant Bioregulators and the Ce
Page 18 and 19:
TOTIPOTENCY AND THE CELL CYCLE 9 in
Page 20 and 21:
TOTIPOTENCY AND THE CELL CYCLE 11 F
Page 22 and 23:
Bartel, D. (2004) MicroRNAs: genomi
Page 24 and 25:
CHAPTER 2 MICROPROPAGATION VIA ORGA
Page 26 and 27:
MICROPROPAGATION OF SLASH PINE Tabl
Page 28 and 29:
MICROPROPAGATION OF SLASH PINE Amon
Page 30 and 31:
2.6. Field Testing MICROPROPAGATION
Page 32 and 33:
CHAPTER 3 MICROPROPAGATION OF COAST
Page 34 and 35:
MICROPROPAGATION OF SEQUOIA SEMPERV
Page 36 and 37: MICROPROPAGATION OF SEQUOIA SEMPERV
Page 42 and 43: CHAPTER 4 MICROPROPAGATION OF PINUS
Page 44 and 45: MICROPROPAGATION OF PINUS PINEA L.
Page 50 and 51: 42 2.1. Organ Culture K. ISHII ET A
Page 52 and 53: 44 K. ISHII ET AL. scent light, 70
Page 54 and 55: 46 Chemicals (mg/l) K. ISHII ET AL.
Page 56 and 57: 48 Table 3. Effects of plant growth
Page 58 and 59: 50 K. ISHII ET AL. Gupta, P.K. & Du
Page 60 and 61: 52 C. HARGREAVES AND M. MENZIES (Me
Page 62 and 63: 54 C. HARGREAVES AND M. MENZIES Con
Page 64 and 65: 56 2.1.3. Organogenesis from Field-
Page 66 and 67: 58 C. HARGREAVES AND M. MENZIES Fig
Page 68 and 69: 60 C. HARGREAVES AND M. MENZIES Fig
Page 70 and 71: 62 C. HARGREAVES AND M. MENZIES For
Page 72 and 73: 64 C. HARGREAVES AND M. MENZIES and
Page 74 and 75: CHAPTER 7 GENETIC FIDELITY ANALYSES
Page 76 and 77: PLANT GENETIC FIDELITY 69 Plant mat
Page 78 and 79: e added to samples, as this fluoroc
Page 80 and 81: 11. Determine the mean channel numb
Page 82 and 83: 3. In addition to testing various b
Page 84 and 85: GeneScan internal size standards: 4
Page 88 and 89: PLANT GENETIC FIDELITY 81 microprop
Page 90 and 91: PLANT GENETIC FIDELITY 83 Steinkell
Page 92 and 93: 86 2.1. Explant Preparation M.G. OS
Page 94 and 95: 88 WPM (Lloyd & McCown, 1980) Macro
Page 96 and 97: 90 M.G. OSTROLUCKÁ ET AL. on the m
Page 98 and 99: CHAPTER 9 MICROPROPAGATION OF MEDIT
Page 100 and 101: 2.1. Explant Preparation MICROPROPA
Page 102 and 103: MICROPROPAGATION OF MEDITERRANEAN C
Page 112 and 113: 108 D.T. NHUT ET AL. Larue (1953) a
Page 114 and 115: 110 D.T. NHUT ET AL. HgCl2 added wi
Page 116 and 117: 112 2.4. Establishment of Shoot Cul
Page 118 and 119: 114 D.T. NHUT ET AL. Table 5. Effec
Page 120 and 121: 116 D.T. NHUT ET AL. culture to be
Page 122 and 123: 118 D. EWALD ability of the plant m
Page 124 and 125: 120 D. EWALD Figure 1. Yew micropro
Page 126 and 127: 122 D. EWALD Root development. Afte
Page 128 and 129: CHAPTER 12 MICROPROPAGATION OF LARI
Page 130 and 131: MICROPROPAGATION OF LARIX SPECIES V
Page 136 and 137:
MICROPROPAGATION OF LARIX SPECIES V
Page 138 and 139:
MICROPROPAGATION OF LARIX SPECIES V
Page 140 and 141:
CHAPTER 13 PROPAGATION OF SELECTED
Page 142 and 143:
PROPAGATION OF SELECTED PINUS GENOT
Page 144 and 145:
Page 146 and 147:
Page 148 and 149:
Page 150 and 151:
CHAPTER 14 ROOT INDUCTION OF PINUS
Page 152 and 153:
ROOT INDUCTION OF PINUS SYLVESTRIS
Page 154 and 155:
ROOT INDUCTION OF PINUS SYLVESTRIS
Page 156 and 157:
CHAPTER 15 MICROPROPAGATION OF BETU
Page 158 and 159:
MICROPROPAGATION OF BETULA PENDULA
Page 160 and 161:
Page 162 and 163:
Page 164 and 165:
Page 166 and 167:
CHAPTER 16 PROTOCOL FOR DOUBLED-HAP
Page 168 and 169:
DOUBLED-HAPLOID MICROPROPAGATION IN
Page 170 and 171:
Page 172 and 173:
Page 174 and 175:
Page 176 and 177:
Page 178 and 179:
Page 180 and 181:
Page 182 and 183:
CHAPTER 17 IN VITRO PROPAGATION OF
Page 184 and 185:
IN VITRO PROPAGATION OF FRAXINUS SP
Page 186 and 187:
Page 188 and 189:
Axillary shoots (number) IN VITRO P
Page 190 and 191:
Page 192 and 193:
Page 194 and 195:
Page 196 and 197:
CHAPTER 18 MICROPROPAGATION OF BLAC
Page 198 and 199:
MICROPROPAGATION OF BLACK LOCUST 19
Page 200 and 201:
2.5. Rooting and Acclimatization MI
Page 202 and 203:
MICROPROPAGATION OF BLACK LOCUST 19
Page 204 and 205:
202 V. RAJESWARI AND K. PALIWAL tha
Page 206 and 207:
204 V. RAJESWARI AND K. PALIWAL Mak
Page 208 and 209:
206 V. RAJESWARI AND K. PALIWAL 2.3
Page 210 and 211:
208 V. RAJESWARI AND K. PALIWAL Fig
Page 212 and 213:
210 2.8. Hardening V. RAJESWARI AND
Page 214 and 215:
CHAPTER 20 MICROPROPAGATION OF SALI
Page 216 and 217:
MICROPROPAGATION OF SALIX CAPREA L.
Page 218 and 219:
Page 220 and 221:
Page 222 and 223:
CHAPTER 21 MICROPROPAGATION OF CEDR
Page 224 and 225:
2.1. Establishment of Shoot Culture
Page 226 and 227:
MICROPROPAGATION OF CEDRELA FISSILI
Page 228 and 229:
Page 230 and 231:
Page 232 and 233:
Page 234 and 235:
Page 236 and 237:
Page 238 and 239:
238 programmes is somatic embryogen
Page 240 and 241:
240 Figure 2. A) Induction of organ
Page 242 and 243:
242 2.4.1. Rooting of Apical and Ba
Page 244 and 245:
244 of donor individuals and/or by
Page 246 and 247:
246 J. MALÀ ET AL. Karnosky, D.F.,
Page 248 and 249:
CHAPTER 23 MICROGRAFTING IN GRAPEVI
Page 250 and 251:
MICROGRAFTING IN GRAPEVINE 251 and
Page 252 and 253:
MICROGRAFTING IN GRAPEVINE 253 Tabl
Page 254 and 255:
2.4. Culture Conditions MICROGRAFTI
Page 256 and 257:
MICROGRAFTING IN GRAPEVINE 257 Feuc
Page 258 and 259:
CHAPTER 24 MICROGRAFTING GRAPEVINE
Page 260 and 261:
MICROGRAFTING GRAPEVINE FOR VIRUS I
Page 262 and 263:
Page 264 and 265:
Page 266 and 267:
CHAPTER 25 APRICOT MICROPROPAGATION
Page 268 and 269:
APRICOT MICROPROPAGATION Figure 1.
Page 270 and 271:
APRICOT MICROPROPAGATION Figure 2.
Page 272 and 273:
APRICOT MICROPROPAGATION 2.2.2. Hyp
Page 274 and 275:
2.4. Acclimatization APRICOT MICROP
Page 276 and 277:
APRICOT MICROPROPAGATION occurs fre
Page 278 and 279:
CHAPTER 26 IN VITRO CONSERVATION AN
Page 280 and 281:
IN VITRO CONSERVATION AND MICROPROP
Page 282 and 283:
Page 284 and 285:
Page 286 and 287:
Page 288 and 289:
CHAPTER 27 MICROGRAFTING OF PISTACH
Page 290 and 291:
MICROGRAFTING OF PISTACHIO Constitu
Page 292 and 293:
MICROGRAFTING OF PISTACHIO 2.2.2. C
Page 294 and 295:
MICROGRAFTING OF PISTACHIO 6. Incub
Page 296 and 297:
MICROGRAFTING OF PISTACHIO 9. The r
Page 298 and 299:
CHAPTER 28 PROTOCOL FOR MICROPROPAG
Page 300 and 301:
MICROPROPAGATION OF CASTANEA SATIVA
Page 302 and 303:
Page 304 and 305:
Page 306 and 307:
Page 308 and 309:
Page 310 and 311:
Page 312 and 313:
CHAPTER 29 MICROPROPAGATION OF CASH
Page 314 and 315:
MICROPROPAGATION OF CASHEW 2.2.1. E
Page 316 and 317:
MICROPROPAGATION OF CASHEW axillary
Page 318 and 319:
MICROPROPAGATION OF CASHEW Table 1.
Page 320 and 321:
MICROPROPAGATION OF CASHEW shade an
Page 322 and 323:
CHAPTER 30 IN VITRO MUTAGENESIS AND
Page 324 and 325:
IN VITRO MUTAGENESIS AND MUTANT MUL
Page 326 and 327:
Page 328 and 329:
Page 330 and 331:
Page 332 and 333:
Page 334 and 335:
336 R.I. IYER compounds (Iyer et al
Page 336 and 337:
A 338 2.2. Culture Medium R.I. IYER
Page 338 and 339:
340 R.I. IYER Figure 2. Formation o
Page 340 and 341:
342 R.I. IYER Figure 3. Formation o
Page 342 and 343:
344 R.I. IYER Rao, Y.S., Mathew, K.
Page 344 and 345:
346 B.K. BISWAS AND S.C. GUPTA marg
Page 346 and 347:
348 B.K. BISWAS AND S.C. GUPTA (iv)
Page 348 and 349:
350 B.K. BISWAS AND S.C. GUPTA 2.4.
Page 350 and 351:
352 B.K. BISWAS AND S.C. GUPTA Figu
Page 352 and 353:
354 B.K. BISWAS AND S.C. GUPTA tree
Page 354 and 355:
356 B.K. BISWAS AND S.C. GUPTA Figu
Page 356 and 357:
358 B.K. BISWAS AND S.C. GUPTA Drew
Page 358 and 359:
CHAPTER 33 MICROPROPAGATION PROTOCO
Page 360 and 361:
MICROPROPAGATION FOR MICROSPORE EMB
Page 362 and 363:
Page 364 and 365:
Page 366 and 367:
Page 368 and 369:
Page 370 and 371:
374 L. TIAN AND S.I. SIBBALD younge
Page 372 and 373:
376 L. TIAN AND S.I. SIBBALD Table
Page 374 and 375:
378 L. TIAN AND S.I. SIBBALD 2.4. R
Page 376 and 377:
CHAPTER 35 MICROPROPAGATION OF JUGL
Page 378 and 379:
MICROPROPAGATION OF JUGLANS REGIA L
Page 380 and 381:
Page 382 and 383:
Page 384 and 385:
Page 386 and 387:
CHAPTER 36 TISSUE CULTURE PROPAGATI
Page 388 and 389:
PROPAGATION OF MONGOLIAN CHERRY AND
Page 390 and 391:
Page 392 and 393:
Page 394 and 395:
Page 396 and 397:
Page 398 and 399:
Page 400 and 401:
Page 402 and 403:
Page 404 and 405:
410 M. PASQUAL AND E.A. FERREIRA 2.
Page 406 and 407:
Page 408 and 409:
414 M. PASQUAL AND E.A. FERREIRA ch
Page 410 and 411:
Page 412 and 413:
418 D.T. NHUT ET AL. its economical
Page 414 and 415:
420 D.T. NHUT ET AL. Table 1. Shoot
Page 416 and 417:
422 D.T. NHUT ET AL. Figure 3. Orga
Page 418 and 419:
424 D.T. NHUT ET AL. mg l -1 NAA +
Page 420 and 421:
426 D.T. NHUT ET AL. Nakasone, H.Y.
Page 422 and 423:
428 C. LIU ET AL. C. cujete L. is c
Page 424 and 425:
430 C. LIU ET AL. Table 1. Composit
Page 426 and 427:
432 C. LIU ET AL. 6. Excise the sho
Page 428 and 429:
434 Fresh weight per plantlet (mg)
Page 430 and 431:
436 C. LIU ET AL. 4. REFERENCES Aga
Page 432 and 433:
438 M. MISHRA ET AL. micropropagati
Page 434 and 435:
440 M. MISHRA ET AL. each plate ino
Page 436 and 437:
Section C
Page 438 and 439:
446 M.G. OSTROLUCKÁ ET AL. from me
Page 440 and 441:
448 M.G. OSTROLUCKÁ ET AL. regener
Page 442 and 443:
450 M.G. OSTROLUCKÁ ET AL. Figure
Page 444 and 445:
452 M.G. OSTROLUCKÁ ET AL. 2.6.3.
Page 446 and 447:
454 counts counts M.G. OSTROLUCKÁ
Page 448 and 449:
CHAPTER 42 PROTOCOL FOR MICROPROPAG
Page 450 and 451:
AN medium (Anderson, 1980) MICROPRO
Page 452 and 453:
MICROPROPAGATION OF VACCINIUM VITIS
Page 454 and 455:
2.6. Flow Cytometry Analysis MICROP
Page 456 and 457:
CHAPTER 43 MICROPROPAGATION OF BAMB
Page 458 and 459:
BAMBOO MICROPROGATION BY AXILLARY S
Page 460 and 461:
Page 462 and 463:
Page 464 and 465:
Page 466 and 467:
Page 468 and 469:
CHAPTER 44 IN VITRO CULTURE OF TREE
Page 470 and 471:
IN VITRO CULTURE OF TREE PEONY 479
Page 472 and 473:
Page 474 and 475:
Page 476 and 477:
Page 478 and 479:
Page 480 and 481:
Page 482 and 483:
Page 484 and 485:
Page 486 and 487:
Page 488 and 489:
Page 490 and 491:
500 M. MHATRE from various natural
Page 492 and 493:
502 2.2. Disinfection of Plant Mate
Page 494 and 495:
504 M. MHATRE soil in polythene bag
Page 496 and 497:
506 M. MHATRE Figure 2. Pineapple,
Page 498 and 499:
508 M. MHATRE Escalona, M., Lorenzo
Page 500 and 501:
510 J.M. AL-KHAYRI Tunisia, Morocco
Page 502 and 503:
512 J.M. AL-KHAYRI 2.1.2. Separatio
Page 504 and 505:
514 2.2. Culture Medium J.M. AL-KHA
Page 506 and 507:
516 J.M. AL-KHAYRI 5. Isolate callu
Page 508 and 509:
518 J.M. AL-KHAYRI 3. Water the tra
Page 510 and 511:
520 J.M. AL-KHAYRI expressed from c
Page 512 and 513:
522 J.M. AL-KHAYRI alcohol and twic
Page 514 and 515:
524 J.M. AL-KHAYRI potential. Simil
Page 516 and 517:
526 J.M. AL-KHAYRI Barreveld, W.H.
Page 518 and 519:
528 D.T. NHUT ET AL. arsenide chip
Page 520 and 521:
530 D.T. NHUT ET AL. Figure 2. Plan
Page 522 and 523:
532 D.T. NHUT ET AL. Figure 4. Plan
Page 524 and 525:
534 D.T. NHUT ET AL. plantlets. The
Page 526 and 527:
536 D.T. NHUT ET AL. Figure 11. Fre
Page 528 and 529:
538 D.T. NHUT ET AL. Figure 13. Sub
Page 530 and 531:
540 D.T. NHUT ET AL. The response o
Page 532 and 533:
CHAPTER 48 IN VITRO MUTAGENESIS IN
Page 534 and 535:
IN VITRO MUTAGENESIS IN BANANA 545
Page 536 and 537:
Page 538 and 539:
Page 540 and 541:
Page 542 and 543:
Page 544 and 545:
3.3. Observations to be Recorded IN
Page 546 and 547:
Page 548:
show all

Protocols for Micropropagation of Woody Trees and Fruits

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?