Manual on MUTATION BREEDING THIRD EDITION Print j-5109 E5 Cover Page 1.indd 1 22.05.18 10:13 Manual on Mutation Breeding Third Edition Edited by Spencer-Lopes, M.M. Forster, B.P. and Jankuloski, L. Plant Breeding and Genetics Subprogramme Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture Vienna, Austria Food and Agriculture Organization of the United Nations International Atomic Energy Agency Vienna, 2018 DISCLAIMER Recommended citation FAO/IAEA. 2018. Manual on Mutation Breeding - Third edition. Spencer-Lopes, M.M., Forster, B.P. and Jankuloski, L. (eds.), Food and Agriculture Organization of the United Nations. Rome, Italy. 301 pp. The designations employed and the presentation of material in this information product do not imply the expression of any opinion whatsoever on the part of the Food and Agriculture Organization of the United Nations (FAO) concerning the legal or development status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. The mention of specific companies or products of manufacturers, whether or not these have been patented, does not imply that these have been endorsed or recommended by FAO in preference to others of a similar nature that are not mentioned. The views expressed in this information product are those of the author(s) and do not necessarily reflect the views or policies of FAO. ISBN 978-92-5-130526-3 © FAO, 2018 FAO encourages the use, reproduction and dissemination of material in this information product. Except where otherwise indicated, material may be copied, downloaded and printed for private study, research and teaching purposes, or for use in non-commercial products or services, provided that appropriate acknowledgement of FAO as the source and copyright holder is given and that FAO’s endorsement of users’ views, products or services is not implied in any way. All requests for translation and adaptation rights, and for resale and other commercial use rights should be made via www.fao.org/contact-us/licence-request or addressed to [email protected]. FAO information products are available on the FAO website (www.fao.org/publications) and can be purchased through publications- [email protected]. Cover photos credits: Shutterstock.com/Tefi, Khakimullin Aleksandr, Zketch, Vitalii Nesterchuk Joint FAO/IAEA Division CONTACT INFORMATION OF THE AUTHORS A. BADIGANNAVAR J.B. JANKOWICZ-CIESLAK Nuclear Agriculture and Biotechnology Plant Breeding and Genetics Laboratory Division FAO/IAEA Agriculture and Biotechnologies Bhabha Atomic Research Center Laboratories Trombay Friedenstrasse 1 400 085, Mumbai, India A-2444 Seibersdorf, Austria Email: [email protected] Email: [email protected] B.P. FORSTER L.JANKULOSKI BioHybrids International Limited Plant Breeding and Genetics Section P.O. Box 2411, Early Joint FAO/IAEA Division Reading RG6 5FY, United Kingdom P.O. Box 100, Wagramer Strasse 5 Email: [email protected]; A-1400 Vienna, Austria [email protected] Email: [email protected] J.S. (Pat) HESLOP-HARRISON P. JANOS University of Leicester Deputy Manager Department of Biology Cereal Research Nonprofit Ltd. Adrian building Alsokikotosor 9. University Road Szeged 6726, Hungary Lei 7RH, Leicester, United Kingdom Email: [email protected] Email: [email protected] A. KODYM R. IBRAHIM University of Vienna Elite Scientific Instruments Sdn. Bhd Core Facility Botanical Garden & A-LG-03, Block A, Serdang Perdana Selatan Department of Pharmacognosy Section 1, 43300 Seri Kembangan Vienna, Austria Selangor, Malaysia Email: [email protected] Email: [email protected] B. KUNTER I. INGELBRECHT Turkish Atomic Energy Authority (TAEK) Plant Breeding and Genetics Laboratory Sarayköy Nuclear Research and Training FAO/IAEA Agriculture and Biotechnologies Centre Laboratories Division of Agriculture, Ankara Turkey Friedenstrasse 1 Email: [email protected] A-2444 Seibersdorf, Austria Email: [email protected] iii M. MATIJEVIC R. SANGWAN Plant Breeding and Genetics Laboratory Université de Picardie Jules Verne UFR FAO/IAEA Agriculture and Biotechnologies Sciences Laboratories Laboratoire EDYSAN-AEB, UMR 7058 Friedenstrasse 1 CNRS A-2444 Seibersdorf, Austria 33, rue Saint Leu, 80000 Amiens France Email: [email protected] Email: [email protected] C. MBA F. SARSU Food and Agriculture Organization of the Plant Breeding and Genetics Section United Nations (FAO) Joint FAO/IAEA Division Viale delle Terme di Caracalla P.O. Box 100, Wagramer Strasse 5 I-00153 Rome, Italy A-1400 Vienna, Austria Email: [email protected] Email: [email protected] A. MUKHTAR ALI GHANIM M. SPENCER-LOPES Plant Breeding and Genetics Laboratory University CAD FAO/IAEA Agriculture and Biotechnologies Faculté des sciences et technologies Laboratories Dept. de biologie végétale Friedenstrasse 1 Dakar, Sénégal A-2444 Seibersdorf, Austria Email: [email protected] Email: [email protected] I. SZAREJKO H. NAKAGAWA Department of Genetics Hamamatsu Photonics K. K. University of Silesia Central Research Laboratory Jagiellonska 28 5000, Hirakuchi, Hamakita-ku 40-032 Katowice, Poland Hamamatsu, Shizuoka, 434-8601 Japan Email: [email protected] Email: [email protected] M. SZURMAN S. NIELEN Faculty of Biology and Environmental Plant Breeding and Genetics Section Protection Joint FAO/IAEA Division University of Silesia P.O. Box 100, Wagramer Strasse 5 Jagiellonska 28 A-1400 Vienna, Austria 40-032 Katowice, Poland Email:[email protected] Email: [email protected] S. PENNA W. THOMAS Nuclear Agriculture and Biotechnology The James Hutton Institute Division Invergowrie Bhabha Atomic Research Institute Dundee DD2 5DA Trombay, Mumbai 400 085, India Scotland, United Kingdom Email: [email protected] Email: [email protected] iv B.J. TILL A. YASSIER University of Vienna Plant Genomics Department of Chromosome Biology Verdant Bioscience Vienna, Austria Galang Email: [email protected] North Sumatra, 20585, Indonesia Email: [email protected] v vi CONTENTS CHAPTER 1. PHYSICAL MUTAGENESIS 5 Spencer-Lopes, M.M., Jankuloski, L., Mukhtar Ali Ghanim, A., Matijevic, M., and Kodym, A. RADIATION TYPES AND SERVICES 5 X-rays 5 Gamma-rays 5 Ultraviolet light 11 Alpha particles 11 Beta particles 12 Particles from accelerators 12 Neutrons 12 Ion beam irradiation and ion beam implantation 13 Cosmic irradiation 14 Laser beam irradiation 14 RADIO-BIOLOGY 14 Absorption of ionizing radiation 14 Chemical effects of ionizing radiation: DNA damage and DNA repair 15 Lethal effects of ionizing radiation: DNA damage and repair 16 DOSIMETRY 16 Exposure and dose determination 17 Absorbed dose in irradiated targets 18 Dosimeters 19 OBJECTIVES AND TREATMENTS 23 Target plant materials 23 Irradiation treatments and conditions 25 RADIATION SENSITIVITY AND MODIFYING FACTORS 28 Environmental factors 28 Biological factors 30 PRE- AND POST-IRRADIATION TREATMENTS 31 Pre-treatment 31 vii Adjustment of seed water content 31 Post-irradiation storage 31 PROTOCOL FOR RADIO-SENSITIVITY TESTING 32 List of equipment, supplies and facilities needed 32 Radio-sensitivity testing procedure 34 STANDARD PROTOCOLS FOR SEED MUTAGENESIS USING X- 44 RAY IRRADIATORS Pre-treatment preparations of seed samples 44 Radiosensitivity tests 44 Irradiation treatment of the seeds 45 Post irradiation treatment and handling 47 PRACTICAL EXAMPLE OF APPLICATIONS USING PHYSICAL 48 MUTAGENS SEED IRRADIATION SERVICES AT THE JOINT FAO/IAEA PLANT 49 BREEDING AND GENETICS LABORATORY OTHER MUTAGENS 49 CHAPTER 2. CHEMICAL MUTAGENESIS 51 Ingelbrecht, I., Jankowicz-Cieslak, J., Szurman, M., Till, B.J., and Szarejko, I. MAIN CHEMICAL MUTAGENS 51 A1kylating agents 52 Sodium azide 53 Other chemical mutagens 54 MODE OF ACTION AND MUTATION SPECTRA 56 Alkylating agents 56 Sodium azide 57 GUIDELINES FOR CHEMICAL MUTAGENESIS 59 Target plant materials 59 Dose, dose determination and mutational load 61 Status of plant materials 62 Physico-chemical properties of chemical mutagens and mutagen solutions 63 Pre- and post-treatment procedures 64 Advantages and limitations of chemical mutagenesis 65 viii STORAGE, HANDLING AND DECONTAMINATION PROCEDURES 66 OF CHEMICAL MUTAGENS Alkyl alkanesulphonates and alkyl sulphates 67 Nitroso compounds 68 Azides 69 EXAMPLES OF TREATMENT PROCEDURES 69 Ethyl methanesulphonate (EMS) mutagenesis of in vitro banana meristem 70 explants Ethyl methanesulphonate (EMS) mutagenesis treatment of barley seeds 74 Combined treatment of barley seeds with sodium azide and N-nitroso N- 77 methylurea Conclusion 81 CHAPTER 3. TYPES OF MUTATIONS 83 Nielen, S., Forster, B.P., and Badigannavar, A. PHENOTYPIC MUTANTS 83 GENOTYPIC MUTATIONS 84 Genome mutations 85 Gene mutations 94 Genes mutations expression at the trait level 95 PRACTICAL EXAMPLES 99 Example 1 - Selection of a single gene mutation: Ug99 rust resistance in 99 wheat Example 2 - Induced mutations for genetic improvement of quantitative 101 traits in sorghum CHAPTER 4. MUTAGEN EFFECTS IN THE FIRST 105 GENERATION AFTER SEED TREATMENT: BIOLOGICAL EFFECTS OF MUTATION TREATMENTS Nielen, S., Forster, B.P., and Heslop-Harrison, J.S. (Pat) PLANT INJURY AND LETHALITY 105 CYTOLOGICAL EFFECTS 107 Chromosome observations 109 The comet assay 109 Low dose stimulation 110 Effects on meiosis 111 ix STERILITY 111 CHIMERAS 112 SECONDARY EFFECTS: TRANSPOSON ACTIVATION 114 Examples of TE-induced mutations in plant breeding 115 CHAPTER 5. MUTATION BREEDING IN SEED PROPAGATED 119 CROPS: PARENTAL SELECTION,