Plant Tissue Culture. Techniques and Experiments PDF
Preview Plant Tissue Culture. Techniques and Experiments
Plait Tissue Culture T e c h n i q u es a nd E x p e r i m e n ts by ROBERTA H. SMITH Department of Soil and Crop Sciences Texas Agricultural Experiment Station Texas A & M University College Station, Texas ACADEMIC PRESS, INC. Harcourt Brace Jovanovich, Publishers San Diego New York Boston London Sydney Tokyo Toronto This book is printed on acid-free paper. (°° Copyright © 1992 by ACADEMIC PRESS, INC. All Rights Reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher. Academic Press, Inc. 1250 Sixth Avenue, San Diego, California 92101-4311 United Kingdom Edition published by Academic Press Limited 24-28 Oval Road, London NW1 7DX Plant Tissue Culture: Techniques and Experiments by Roberta H. Smith International Standard Book Number 0-12-650340-0 PRINTED IN THE UNITED STATES OF AMERICA 92 93 94 95 96 97 KP 9 8 7 6 5 4 3 2 1 To my parents Preface This manual resulted from the need for plant tissue culture laboratory exercises that demonstrate major concepts and that use plant material that is available year round. Each chapter on the main plant cell culture techniques has plant material options for selecting the explant source, including seed catalogs, plant nurseries, feed stores, landscape plant materials, and produce departments at grocery stores. In short, the strategy in developing this manual has been to de- vise exercises that do not require maintenance of an exten- sive collection of plant materials, and to give students the opportunity to work with such materials. In that respect, this manual is different from those focusing on carrot or tobacco cell culture systems. The exercises in this manual have been used by a wide range of students, from high school (science fair and 4-H projects) to undergraduate and graduate levels. The manual is predominantly directed at upper-level college or university classes in which the students have taken courses in chemis- try, plant anatomy, and plant physiology. Before starting the exercises the student should exam- ine Chapters 1-3, which deal with the setup of a tissue cul- ture laboratory, media preparation, and explants and aseptic XI xii Preface technique. The information in these chapters will be needed in the exercises that follow. The brief introduction to each chapter is not intended to be a review of the chapter's topic but rather a complement to lecture discussions of the topic. In many instances, plant material initiated in one exercise is used in subsequent exercises. Refer to Scheduling and Interrelationships of Exercises to obtain information on the time required to complete the exercises and how they relate to one another. All of the exercises have been successfully accomplished for at least two semesters, and most have been successfully used for five semesters. Tissue culture, however, is still more art than science, and variation in individual exercises can be expected. Roberta H. Smith Attainments I would like to acknowledge the teaching assistants who helped in developing some of these exercises: Daniel Caulkins, Cheryl Knox, John Finer, Richard Norris, Ann Reilley- Panella, Ricardo Diquez, Eugenio Ulian, Shelley Gore, Sara Perez-Ramos, Jeffrey Callin, and Greg Peterson. Additionally, all the students who have taken my tissue culture course since 1979 have been important in developing and improv ing these exercises. Kay Hesby is acknowledged for proofread ing the manuscript. Linda Kocman, in particular, was important in the proofreading, formatting, and typing of the many versions of this manual. XIII 'ίιιΐιπΐιιΐιιι;: and Interrelationships of Exercises I. Aseptic Germination of Seed (Chapter 3) Carrot: 1-2 wk; Cotton, Sunflower: 1 wk A. Callus Induction (Chapter 4): 6 wk Broccoli, Lemon: 6 wk Carrot: 2 subcultures, 6 wk each = 3 mo 1. Salt Selection in Vitro (Chapter 4): 4 wk 2. Suspension Culture (Chapter 5): 2 wk Carrot a. Somatic Embryogenesis (Chapter 5): 3-4 wk b. Explant Orientation (Chapter 4): 6 wk Cotton: 2 subcultures, 6 wk each 1. Protoplast (Chapter 10): 2 days 2. Cellular Variation (Chapter 4): 4 wk 3. Growth Curves (Chapter 4): 6 wk II. Tobacco Seed Germination (Chapter 4): 3 wk A. Callus Induction (Chapter 4): 2 subcultures, 6 wk each xv xvi Scheduling and Interrelationships of Exercises III. Establishment of Competent Cereal Cell Cultures (Chapter 4): 2-3 wk A. Rice Subculture (Chapter 5): 3 wk 1. Plant Regeneration: 4 wk IV. Potato Shoot Initiation (Chapter 5): 6 wk A. Potato Tuberization (Chapter 5): 4-6 wk V. Douglas Fir Seed Germination (Chapter 3): 2-4 wk A. Primary Morphogenesis (Chapter 5): 4 wk VI. Petunia Leaf Disk Transformation (Chapter 11): 6 wk VII. Petunia Shoot Apex Transformation (Chapter 11): 4-6 wk VIII. Solitary Exercises A. Bulb Scale Dormancy (Chapter 5): 6-8 wk B. Datura Anther Culture (Chapter 6): 4-8 wk; 10 wk to obtain flowering plants C. African Violet Anther Culture (Chapter 6): 7-8 wk D. Tobacco Anther Culture (Chapter 6): 7-8 wk; 2-3 mo to obtain flowering plants E. Corn Embryo Culture (Chapter 7): 72 hr F. Crabapple and Pear Embryo Culture (Chapter 7): 2-3 wk G. Shoot Apical Meristem (Chapter 8): 4-6 wk H. Diffenbachia Meristem (Chapter 8): 4-6 wk I. Garlic Propagation (Chapter 8): 4 wk J. Boston Fern Propagation (Chapter 9) Stage I: 6-8 wk Stage II: 4-6 wk Stage III: 2-3 wk K. Staghorn Fern Propagation (Chapter 9) Stage I: 2-3 wk Stage II: 6 wk Stage III: 4-6 wk Scheduling and Interrelationships of Exercises xvil L. Ficus Propagation (Chapter 9) Stage I: 4-6 wk Stage II: 4-6 wk Stage III: 4 wk M. Kalanchoe Propagation, Stages I & II (Chapter 9): 4 wk N. Rose Propagation, Stages I & II (Chapter 9): 8 wk 0. African Violet, Stages I & II (Chapter 9): 4 wk P. Pitcher Plant, Stages I & II (Chapter 9): 6 wk Q. Cactus Propagation (Chapter 9) Stage I: 4-6 wk Stage II: 4-6 wk Stage III: 8 wk D Setup of a Tissue Culture Laboratory The tissue culture area should be kept picked up and clean at all times. This is importan t to ensure clean cultures and reproducible results. Avoid having potted plants in this area becaus e they can be a source of mites and other contami - nating organisms . Avoid field or greenhous e work immediatel y before entering the laborator y becaus e mites and insects can be carried into the laborator y on hair and clothing. Shower and change clothes before entering the laborator y from the field or greenhouse . Determining the location of the tissue culture laborator y is an importan t decision. Avoid locating it adjacen t to labora- tories that handle microorganism s or insects or are used to store seed s or other plant materials . Contaminatio n from air vents and traffic can be a problem. In designing a laborator y for tissue culture use, arrange the work areas (media preparation , culture transfer , and 1
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