Table Of ContentMicropropagation of Orchids
Volume I
This, the third and expanded and updated edition of a classic reference work, provides users with
comprehensive details of all procedures developed by May 2014 for orchid propagation through the
culture of tissues in vitro (micropropagation).
The book opens with an extensive, expanded with new information, and richly illustrated historical
chapter which discusses the discoveries, developments, and people that made micropropagation possible.
An updated and expanded Chapter 2 discusses in detail the principles and components of culture media
used in micropropagation. However, the main component of these volumes is the third chapter, which
contains detailed procedures for the culture and micropropagation of hundreds of orchid genera, species,
and hybrids. Early, classic, established, and the very latest techniques of mass rapid clonal propagation
through the culture of organs, tissues, and cells are presented. Every one of the methods in Chapter 3
includes tables of complete recipes for a wide range of culture media.
This new edition will continue to be the key reference for all those interested or involved in orchid micro-
propagation, growing, and production.
Dr Yam Tim Wing studied Biology (B.Sc.) at the University of Southampton in England. He received his
M.Phil. degree in plant breeding and genetics from Cambridge University before returning to Hong Kong
where he earned his Ph.D. with a research project on the conservation of the native orchids. From 1988
to 1991, he carried out postdoctoral research at the University of California, Irvine with Professor Joseph
Arditti. He is a Principal Researcher at the Singapore Botanic Gardens, National Parks Board, specializing
in orchid hybridization and conservation. His hybrids have won awards at local and international shows.
He also administers a program designed to conserve the native orchids of Singapore by propagating and
introducing these species into urban and natural areas in the country. Dr. Yam is the author of research
papers and two books entitled Orchids of the Singapore Botanic Gardens and Native Orchids of Singapore:
Identification, Diversity and Conservation. He also co‐authored the book Biology of Vanda Miss Joaquim
with Professors Hew Choy Sin (Western style: Choy sin Hew) and Joseph Arditti. Dr. Yam is married to
Pauline Wong and they have two daughters, Grace and Amy.
Professor Emeritus Joseph Arditti majored in floriculture as an undergraduate and received his doctorate
from the University of Southern California in 1965. After serving as a lecturer for one year at the University
of Southern California, he accepted a faculty position at the University of California, Irvine in 1966 where
he taught general botany, horticulture, and plant physiology while engaging in research on various aspects
of orchid biology in the USA, Indonesia, Singapore, and Malaysia. Dr. Arditti retired in 2001 and is now
Professor of Biology Emeritus. He continued to write and travel worldwide to lecture at scientific meetings
and to orchid groups until 2014 when he retired for a second time. Professor Arditti is acknowledged to be
one of the world’s leading experts on orchid biology and propagation. He is the proud father of Dr. (foren-
sic psychology) Jonathan O. Arditti who received his bachelor’s degree of psychology from the University
of Southern California and equally proud father in law to Dr. Alexandria Grabowski who also received a
bachelor’s degree and a doctorate in dentistry from the same university.
Micropropagation of Orchids
Third Edition
Volume I
Tim Wing Yam
Singapore Botanic Gardens
National Parks Board
Singapore
and
Joseph Arditti
Professor Emeritus
Developmental and Cell Biology
University of California, Irvine
This edition first published 2017
© 2017 by John Wiley & Sons Ltd
Edition history: John Wiley & Sons, Inc. (1e, 1993); Blackwell Publishing Ltd (2e, 2008)
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Library of Congress Cataloging‐in‐Publication Data
Names: Yam, Tim Wing. | Arditti, Joseph.
Title: Micropropagation of orchids / Tim Wing Yam and Joseph Arditti.
Description: Third edition. | Chichester, West Sussex : John Wiley & Sons, Inc., 2017. |
Previous edition: Micropropagation of orchids / Joseph Arditti (Malden, MA ; Oxford :
Blackwell Pub., 2008). | Includes bibliographical references and index.
Identifiers: LCCN 2016043344 | ISBN 9781119187059 (cloth) | ISBN 9781119187073 (epub)
Subjects: LCSH: Orchids–Micropropagation.
Classification: LCC SB409.58 .A73 2017 | DDC 584/.4–dc23
LC record available at https://lccn.loc.gov/2016043344
A catalogue record for this book is available from the British Library.
Wiley also publishes its books in a variety of electronic formats. Some content that appears
in print may not be available in electronic books.
Cover images courtesy of Tim Wing Yam.
Cover design by Wiley
Set in 10/12pt Sabon by SPi Global, Pondicherry, India
1 2017
For
My wife Pauline (center) and my daughters Grace The memory of my doctoral
(left) and Amy (holding the family dog, Powdy). studies supervisor,
Dr. Maureen Weatherhead
Tim Wing Yam
My son, Dr. Jonathan Arditti, and my daughter in
law, Dr. Alexandria (Alex) Arditti (born
Grabowski), both alumni of the University of
Mike and Anne Grabowski, alumni of the
Southern California, like the old man.
University of Southern California, like their
daughter.
Joseph Arditti
Short Contents
Full Contents for Volume I, ix
Preface, xiii
Preface to the Second Edition, xvi
Preface to the First Edition, xxi
VOLUME I
Chapter 1: History, 1
Chapter 2: General Outline of Techniques and Procedures, 73
Chapter 3: Methods for Specific Genera, Acampe to Darwinara, 169
VOLUME II
Chapter 3: Methods for Specific Genera, Dendrobium to Phalaenopsis, 699
VOLUME III
Chapter 3:Methods for Specific Genera, Pholidota to Zygopetalum, 1561
Chapter 4: Summary, 1928
References, 1965
Appendix 1: General Information on Supplies, Equipment, Terms,
and Reagents, 2061
Appendix 2: Sources of Supplies and Equipment, 2080
Appendix 3: Some Sites of Interest on the World Wide Web, 2085
Appendix 4: Light, 2088
Appendix 5: Formulary, 2093
vii
viii Short Contents
Appendix 6: Atomic Weights, Concentrations, Exponents, Greek and Roman Letters,
Ions, Measurements, Molecular Weights, Prefixes, Valences, Units,
and Solutions, 2106
Appendix 7: Additional Information, 2116
Appendix 8: Plant Preservative Mixture, 2124
Appendix 9: Media: Additives, Components, Errors, Modifications, Omissions,
Substitutions, Sterilization and More, 2138
Appendix 10: Computer Letters and Symbol Codes, 2150
Appendix 11: Photographing Cultures in Flasks, 2155
Glossary, 2157
General Subject Index, 2180
Full Contents for Volume I
Preface, xiii
Preface to the Second Edition, xvi
Preface to the First Edition, xxi
Chapter 1: History, 1
Terminology, 2
Origins of Orchid Micropropagation, 5
Plant Hormones and Propagation Additives of Plant Origin, 5
Culture of Tissues and Organs, 10
Early Tissue Culture Attempts, 11
Culture of Stem Tips, 12
First Micropropagation of Orchids, 18
The Second Aseptic Culture of an Orchid Explant, 22
Plant Diseases and Meristems, 27
The Third Aseptic Culture of an Orchid Explant, 30
The Fourth Aseptic Culture of an Orchid Explant, 45
Who Pioneered Micropropagation?, 47
Root Cultures, 48
Leaf Cultures, 50
Stems, 51
Flower Buds, Flowers, Floral Segments, and Reproductive Organs, 52
Inflorescences, 52
A Patent, 53
Doubtful Claims, 55
Mutations, 56
Theft in Vitro, 57
Darkening of Culture Media, 57
Anticontaminants, 58
Cell and Protoplast Culture, 59
Flowering In Vitro, 62
ix
x Contents
Solidifying Agents, 65
Agar, 66
Gellan Gum, 68
The Future, 70
Chapter 2: General Outline of Techniques and Procedures, 73
Media Components, 73
Macroelements, 74
Microelements, 83
Iron, 83
Auxins and Anti‐auxin, 84
Chitosan, 88
Cytokinins, 88
Gibberellins and Antigibberellin, 89
Abscisic Acid, 89
Ethylene and Ethylene Inhibitor, 90
Amino Acids, 90
Polyol, 90
Polyamines, 91
Phloroglucinol, 91
Jasmonates, 91
Herbicides, 91
Vitamins, 91
Nucleotides and Nucleic Acids, 92
Organic Acids, 92
Banana, 92
Coconut Water, 92
Humic acid, 98
Hydrolysates and Autolysates, 98
Sugars, 98
Anticontaminants, 103
Charcoal, 110
Solvents, 114
Solidifiers, 114
pH, 120
Recipes in this Book, 121
Stock Solutions, 121
State of the Medium, 122
Sterilization, 122
Autoclaves, 122
Filtration, 123
Sterilization in Microwave Ovens, 124
Solvents, 126
Open Flame, 126
Liquids, 127
Contents xi
Surface Decontamination, 128
Gas Sterilization, 129
Preparation of the Medium, 130
Culture Vessels, 131
Culture Conditions, 132
Temperature, 132
Agitation, 132
Illumination, 133
Carbon Dioxide, 148
Placing Plant Material in Culture, 151
Seeds, 151
Tissue Explants, 151
Internal Contaminants, 155
Mites and Thrips as Contaminants and Contamination Vectors, 156
Hygiene, 158
Prevention, 159
Culture and Encapsulation Media, 161
Fumigation, 161
Eradication, 162
Thrips, 163
Springtails, 163
Virus infections and testing, 164
Enzymes for Protoplast Isolation, 164
Work Area, 166
Laminar‐flow Hoods, 166
Sterile Rooms, 166
Sterile Boxes, 167
Clean Laboratory Bench, 167
Washing Glassware, 167
Statistics, 168
Chapter 3: Methods for Specific Genera, 169
Acampe, 169
Aeridachnis, 185
Aerides, 186
Aeridovanda, 208
Anacamptis, 209
Angraecum, 213
Anoectochilus, 217
Ansellia, 235
Arachnis, 239
Arachnostylis, 249
Aranda, 250
Aranthera, 278
Arundina, 287
