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Agrobacterium Protocols M E T H O D S I N M O L E C U L a R B I O L O G Y™ Agrobacterium Protocols M E T H O D S I N M O L E C U L A R B I O L O G Y™ John M. Walker, SERIES EDITOR 371. Biological Aging: Methods and Protocols, edited by 349349. YAC Protocols, Second Edition, edited by Alasdair Trygve O. Tollefsbol, 2007 MacKenzie, 2006 370. Adhesion Protein Protocols, Second Edition, edited 348348. Nuclear Transfer Protocols: Cell Reprogramming by Amanda S. Coutts, 2007 and Transgenesis, edited by Paul J. Verma and Alan 369. Electron Microscopy: Methods and Protocols, Trounson, 2006 Second Edition, edited by John Kuo, 2007 347.347 Glycobiology Protocols, edited by Inka Brockhausen- 368. Cryopreservation and Freeze-Drying Protocols, Schutzbach, 2006 Second Edition, edited by John G. Day and Glyn 346.346 Dictyostelium discoideum Protocols, edited by Stacey, 2007 Ludwig Eichinger and Francisco Rivero-Crespo, 2006 367. Mass Spectrometry Data Analysis in Proteomics, 345.345 Diagnostic Bacteriology Protocols, Second Edition, edited by Rune Mattiesen, 2007 edited by Louise O'Connor, 2006 366. Cardiac Gene Expression: Methods and Protocols, 344.344 Agrobacterium Protocols, Second Edition: edited by Jun Zhang and Gregg Rokosh, 2007 Volume 2, edited by Kan Wang, 2006 365. Protein Phosphatase Protocols: edited by Greg 343.343 Agrobacterium Protocols, Second Edition: Moorhead, 2007 Volume 1, edited by Kan Wang, 2006 364. Macromolecular Crystallography Protocols: 342.342 MicroRNA Protocols, edited by Shao-Yao Ying, 2006 Volume 2, Structure Determination, edited by Sylvie 341.341 Cell–Cell Interactions: Methods and Protocols, Doublié, 2007 edited by Sean P. Colgan, 2006 363. Macromolecular Crystallography Protocols: 340.340 Protein Design: Methods and Applications, Volume 1, Preparation and Crystallization edited by Raphael Guerois and Manuela López de la of Macromolecules, edited by Sylvie Doublié, 2007 Paz, 2006 362. Circadian Rhythms: Methods and Protocols, 339.339 Microchip Capillary Electrophoresis: Methods edited by Ezio Rosato, 2007 and Protocols, edited by Charles S. Henry, 2006 361. Target Discovery and Validation Reviews 338.338 Gene Mapping, Discovery, and Expression: and Protocols: Emerging Molecular Targets Methods and Protocols, edited by M. Bina, 2006 and Treatment Options, Volume 2, edited by 337.337 Ion Channels: Methods and Protocols, edited by Mouldy Sioud, 2007 James D. Stockand and Mark S. Shapiro, 2006 360. Target Discovery and Validation Reviews 336.336 Clinical Applications of PCR, Second Edition, and Protocols: Emerging Strategies for Targets edited by Y. M. Dennis Lo, Rossa W. K. Chiu, and K. C. and Biomarker Discovery, Volume 1, edited by Allen Chan, 2006 Mouldy Sioud, 2007 335.335 Fluorescent Energy Transfer Nucleic Acid 359. Quantitative Proteomics, edited by Salvatore Sechi, Probes: Designs and Protocols, edited by Vladimir 2007 V. Didenko, 2006 358. Metabolomics: Methods and Protocols, edited by 334.334 PRINS and In Situ PCR Protocols, Second Wolfram Weckworth, 2007 Edition, edited by Franck Pellestor, 2006 357. Cardiovascular Proteomics: Methods and Protocols, 333.333 Transplantation Immunology: Methods and edited by Fernando Vivanco, 2006 Protocols, edited by Philip Hornick and Marlene Rose, 2006 356. High-Content Screening: A Powerful Approach to Systems Cell Biology and Drug Discovery, edited by 332.332 Transmembrane Signaling Protocols, Second Ken Guiliano, D. Lansing Taylor, and Jeffrey Haskins, Edition, edited by Hydar Ali and Bodduluri Haribabu, 2006 2006 355. Plant Proteomics: Methods and Protocols, edited 331331. Human Embryonic Stem Cell Protocols, edited by by Hervé Thiellement, Michel Zivy, Catherine Kursad Turksen, 2006 Damerval, and Valerie Mechin, 2006 330330. Embryonic Stem Cell Protocols, Second Edition, 354. Plant–Pathogen Interactions: Methods and Volume II: Differentiation Models, edited by Kursad Protocols, edited by Pamela C. Ronald, 2006 Turksen, 2006 353. DNA Analysis by Nonradioactive Probes: Methods 329329.Embryonic Stem Cell Protocols, Second Edition, and Protocols, edited by Elena Hilario and John. F. Volume I: Isolation and Characterization, edited by MacKay, 2006 Kursad Turksen, 2006 32328.8 New and Emerging Proteomic Techniques, edited 35352.2 Protein Engineering Protocols, edited by Kristian Müller and Katja Arndt, 2006 by Dobrin Nedelkov and Randall W. Nelson, 2006 327.327 Epidermal Growth Factor: Methods and Protocols, 351351. C. elegans: Methods and Applications, edited by Kevin Strange, 2006 edited by Tarun B. Patel and Paul J. Bertics, 2006 326.326 In Situ Hybridization Protocols, Third Edition, 350350. Protein Folding Protocols, edited by Yawen Bai and Ruth Nussinov 2006 edited by Ian A. Darby and Tim D. Hewitson, 2006 M E T H O D S I N M O L E C U L A R B I O L O G Y™ Agrobacterium Protocols Second Edition Volume 1 Edited by Kan Wang Center for Plant Transformation, Plant Science Institute, and Department of Agronomy, Iowa State University, Ames, Iowa © 2006 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 www.humanapress.com All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Molecular BiologyTM is a trademark of The Humana Press Inc. All papers, comments, opinions, conclusions, or recommendations are those of the author(s), and do not necessarily reflect the views of the publisher. This publication is printed on acid-free paper. f ANSI Z39.48-1984 (American Standards Institute) Permanence of Paper for Printed Library Materials. Cover design by Patricia F. Cleary Cover illustration: Agrobacterium cells attaching to plant tissue surface. Artwork provided by Dr. Kan Wang. For additional copies, pricing for bulk purchases, and/or information about other Humana titles, contact Humana at the above address or at any of the following numbers: Tel.: 973-256-1699; Fax: 973-256-8341; E-mail: [email protected]; or visit our Website: www.humanapress.com Photocopy Authorization Policy: Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Humana Press Inc., provided that the base fee of US $30.00 per copy is paid directly to the Copyright Clearance Center at 222 Rosewood Drive, Danvers, MA 01923. For those organizations that have been granted a photocopy license from the CCC, a separate system of payment has been arranged and is acceptable to Humana Press Inc. The fee code for users of the Transactional Reporting Service is: [1-58829-536-2/06 $30.00 ]. Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 EISBN 1-59745-130-4 Library of Congress Cataloging in Publication Data Agrobacterium protocols / edited by Kan Wang.— 2nd ed. p. ; cm. — (Methods in molecular biology ; 343) Includes bibliographical references and index. ISBN 1-58829-536-2 (alk. paper) 1. Agrobacterium. 2. Agricultural biotechnology. [DNLM: 1. Rhizobium radiobacter—genetics—Laboratory Manuals. 2. Rhizobium radiobacter—isolation & purification—Laboratory Manuals. 3. Bacteriological Techniques—methods—Laboratory Manuals. 4. Genetic Techniques—Laboratory Manuals. QW 25 A281 2006] I. Wang, Kan, 1957- II. Series: Methods in molecular biology (Clifton, N.J.) ; v. 343. QR82.R45A37 2006 631.5’233—dc22 2005029677 To Marc Van Montagu and Jeff Schell (1935–2003), my PhD mentors, for their inspiration and encouragement. Preface Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agricultural development. Compared to physical transformation methods such as particle bombard- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate a single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Agrobacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent developments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells. While the list of organisms that can be infected by Agrobacterium has increased significantly over the past decade, the success in transformation also relies on culture responsiveness of the target cells/tissues subsequent to the co-cultivation with Agrobacterium. Essentially, the dynamic interactions between the two living organisms are critical for development of transforma- tion methods. The second edition of Agrobacterium Protocols contains 80 chapters (two volumes) divided into 14 parts. Part I in Volume 1 (Agrobacterium Handling) provides six chapters describing basic techniques in Agrobacterium manipula- tion and strategies for vector construction,
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