Biotechnologies for Plant Mutation Breeding : : Protocols.

• Author: Jankowicz-Cieslak, Joanna.
• TeilnehmendeR: Tai, Thomas H.; Kumlehn, Jochen.; Till, Bradley J.
• Place / Publishing House: Cham : : Springer International Publishing AG,, 2016.; Ã2017.
• Year of Publication: 2016
• Edition: 1st ed.
• Language: English
• Physical Description: 1 online resource (343 pages)

Table of Contents

Intro -- Preface -- Acknowledgements -- Contents -- Chapter Reviewers -- Contributors -- Part I: Introduction -- Chapter 1: Mutagenesis for Crop Breeding and Functional Genomics -- 1.1 Inducing Genetic Variation -- 1.1.1 Practical Considerations in Induced Crop Mutagenesis -- 1.1.2 Developing Crop Varieties Using Induced Mutations -- 1.1.3 Elite Crop Varieties Developed Through Induced Mutations -- 1.2 Phenotypic Screening -- 1.2.1 Phenotypic Traits Developed Through Plant Mutation Breeding -- 1.3 Genotypic Screening of Mutant Plants -- 1.3.1 Genotypic Methods -- 1.3.1.1 Lower-Cost Mutation Discovery and Genotyping Methods -- 1.3.1.2 Higher-Throughput Genotyping and Mutation Discovery Methods -- 1.3.1.3 Cloning Mutant Alleles Causative for Improved Traits -- 1.4 Conclusion -- References -- Part II: Mutation Induction and Chimera Dissociation -- Chapter 2: Chemical and Physical Mutagenesis in Jatropha curcas -- 2.1 Introduction -- 2.2 Materials -- 2.2.1 In Vivo Material -- 2.2.2 In Vitro Material -- 2.2.3 Mutagenesis by Chemical Agents (See Note 2) -- 2.2.4 Mutagenesis by Physical Agents -- 2.3 Methods -- 2.3.1 In Vivo Material -- 2.3.2 In Vitro Material -- 2.3.3 Mutagenesis by Chemical Agents -- 2.3.3.1 EMS Mutagenesis of In Vivo Material (See Note 10) -- 2.3.3.2 EMS Mutagenesis of In Vitro Material (See Notes 10 and 20 and Fig. 2.2) -- 2.3.4 Mutagenesis by Physical Agents -- 2.3.4.1 Gamma Irradiation of In Vivo Material (See Notes 21-22) -- 2.3.4.2 Gamma Irradiation of In Vitro Material -- 2.3.4.3 X-Rays (See Note 32, Fig. 2.4) -- 2.4 Further Analyses -- 2.5 Notes -- References -- Chapter 3: Chemical Mutagenesis and Chimera Dissolution in Vegetatively Propagated Banana -- 3.1 Introduction -- 3.2 Materials -- 3.2.1 Culture Medium (S-27) -- 3.2.2 Chemical Toxicity Test -- 3.2.3 Calculation of Growth Reduction (GR) -- 3.2.4 Bulk Mutagenesis.

3.2.5 Chimera Dissolution -- 3.3 Methods -- 3.3.1 Preparation of Liquid Culture Medium -- 3.3.2 Preparation of Solid Culture Medium -- 3.3.3 Chemical Toxicity Test -- 3.3.4 Calculation of Growth Reduction (GR) -- 3.3.5 Bulk Mutagenesis -- 3.3.6 Chimera Dissolution -- 3.4 Notes -- References -- Chapter 4: Mutation Induction Using Gamma Irradiation and Embryogenic Cell Suspensions in Plantain (Musa spp.) -- 4.1 Introduction -- 4.1.1 Somatic Embryogenesis in Musa spp. -- 4.1.2 Mutation Induction in Musa spp. -- 4.2 Materials -- 4.2.1 Explant Preparation: Shoot-Tip Establishment and Multiplication -- 4.2.2 Culture Medium and Incubation Materials -- 4.2.3 Acclimatization -- 4.2.4 Mutation Induction Using Gamma Irradiation -- 4.3 Methods -- 4.3.1 Explant Preparation: Shoot-Tip Establishment and Multiplication -- 4.3.1.1 Shoot-Tip Establishment -- 4.3.1.2 Shoot-Tip Multiplication -- 4.3.2 Protocol for Plant Regeneration via Somatic Embryogenesis -- 4.3.2.1 Callus Formation with Embryogenic Structures -- 4.3.2.2 Establishment and Multiplication of Embryogenic Cell Suspensions -- 4.3.2.3 Formation of Somatic Embryos -- 4.3.2.4 Maturation of Somatic Embryos -- 4.3.2.5 Germination of Somatic Embryos -- 4.3.2.6 Acclimatization Phase of Somatic Embryos, Conversion into Plants -- 4.3.3 Mutation Induction Using Gamma Irradiation -- 4.4 Notes -- References -- Chapter 5: Optimisation of Somatic Embryogenesis in Cassava -- 5.1 Introduction -- 5.1.1 Production of Cyclic Embryos -- 5.1.2 Influence of Growth Regulators on Primary Embryo Induction -- 5.1.3 Somatic Embryo Conversion into Plants -- 5.2 Materials -- 5.2.1 Chemicals and Equipment -- 5.2.2 Culture Media -- 5.2.3 Shoot Initiation Medium (see Notes 2-4) -- 5.2.4 Embryo Initiation Medium -- 5.2.5 Embryo Maturation Medium -- 5.2.6 Somatic Embryo Conversion Medium -- 5.3 Methods.

5.3.1 Preparation of Shoot Initiation Medium -- 5.3.2 Collection and Sterilisation of Donor Plants -- 5.3.3 Sterilisation and Culture for Shoot Initiation -- 5.3.4 Culture Incubation -- 5.3.5 Initiation of Primary Somatic Embryos -- 5.3.6 Cyclic Embryo Initiation and Production -- 5.3.7 Abscisic Acid Effect on Conversion of Somatic Embryos into Plant -- 5.3.8 Desiccation of Embryos for Plant Conversion -- 5.4 Notes -- 5.5 Conclusion -- References -- Chapter 6: Creation of a TILLING Population in Barley After Chemical Mutagenesis with Sodium Azide and MNU -- 6.1 Introduction -- 6.2 Materials -- 6.2.1 Mutagenesis -- 6.2.2 Handling of Mutated Population -- 6.2.3 DNA Isolation -- 6.2.4 Creation of a Database -- 6.3 Methods -- 6.3.1 Mutagenesis -- 6.3.1.1 General Remarks -- 6.3.1.2 Mutagenic treatment -- 6.3.1.3 Evaluation of a Critical Dose of Mutagens -- 6.3.2 Handling of the Mutated Generations and the Basic Phenotyping of M2 Plants and M3 Lines -- 6.3.3 DNA Isolation: Creating the M2 DNA Library -- 6.3.3.1 Isolate DNA from the M2 Plants According to the Modified Micro-CTAB Method (Doyle and Doyle 1987 and see Note 19) -- 6.3.3.2 Prepare Pools of DNA to Identify Plants Carrying Mutations Within the Gene of Interest -- 6.3.4 Creation of a Database -- 6.3.4.1 General Information -- 6.3.4.2 Creation of a Database: An Example -- Notes -- References -- Chapter 7: Site-Directed Mutagenesis in Barley by Expression of TALE Nuclease in Embryogenic Pollen -- 7.1 Introduction -- 7.1.1 Site-Directed Mutagenesis in Plants -- 7.1.2 Haploid Technology -- 7.2 Materials -- 7.2.1 Donor Plants -- 7.2.2 Stock Solutions and Culture Media -- 7.2.2.1 Stock Solutions -- 7.2.2.2 Medium for Agrobacterium Tumefaciens -- 7.2.2.3 Media for Plant Cell Culture -- 7.2.3 Materials for the Isolation of Embryogenic Pollen -- 7.2.4 Materials for Agrobacterium-Mediated Transformation.

7.2.5 Materials for the Analysis of Transgenic Plants -- 7.2.5.1 Ploidy Determination and Colchicine-Induced Whole Genome Duplication -- 7.2.5.2 Molecular Analyses -- DNA Isolation, PCR, and DNA Gel Blot Analysis -- RNA Isolation and Reverse Transcriptase Reaction -- 7.3 Methods -- 7.3.1 Vector Construction and Bacterial Strains -- 7.3.2 Growth of Donor Plants -- 7.3.3 Isolation of Immature Pollen -- 7.3.3.1 Spike Pretreatment -- 7.3.3.2 Isolation, Purification, and Pre-cultivation of Immature Pollen -- 7.3.4 Agrobacterium-Mediated Gene Transfer to Embryogenic Pollen -- 7.3.4.1 Preparation of A. tumefaciens Stocks -- 7.3.4.2 Cocultivation of Embryogenic Pollen Cultures and A. tumefaciens -- 7.3.5 Regeneration of Transgenics -- 7.3.6 Analysis of Putative Transgenic Plants -- 7.3.6.1 Ploidy Determination and Colchicine-Induced Whole Genome Duplication -- 7.3.6.2 Molecular Analyses -- DNA Isolation, PCR, and DNA Gel Blot Analysis -- RNA Isolation and Reverse Transcriptase Reaction -- 7.4 Notes -- References -- Chapter 8: Doubled Haploidy as a Tool for Chimaera Dissolution of TALEN-Induced Mutations in Barley -- 8.1 Introduction -- 8.1.1 Generation of Primary Mutants by Cross-Combination of Parental Lines Carrying Complementary Single TALEN Units -- 8.1.2 Chimaerism Upon TALEN-Induced Targeted Mutagenesis -- 8.1.3 Haploid Technology -- 8.2 Materials -- 8.2.1 Growth of Parental Lines and Primary Mutant Plants -- 8.2.2 Stock Solutions and Culture Media -- 8.2.2.1 Solutions for Isolation, Purification and Induction of Embryogenic Development of Immature Pollen -- 8.2.2.2 Nutrient Media for Embryogenic Pollen Culture and Plant Regeneration -- 8.2.3 Materials for the Isolation of Embryogenic Pollen Culture and Plant Regeneration -- 8.2.4 Materials for the Molecular Analyses of Pollen-Derived Plants.

8.2.5 Materials for Ploidy Determination and Colchicine-Induced Whole-Genome Duplication -- 8.3 Methods -- 8.3.1 Growth of Primary Mutant Plants -- 8.3.2 Crossing of Pairs of Complementary Single TALEN Plants and Analysis of Hybrid Plants -- 8.3.3 Spike Preparation -- 8.3.4 Isolation, Purification and Inductive Treatment of Immature Pollen -- 8.3.5 Regeneration of Pollen-Derived Plants -- 8.3.6 Analysis of Pollen-Derived Plants -- 8.3.6.1 Ploidy Determination -- 8.3.6.2 DNA Isolation, PCR and Sequencing -- 8.3.6.3 Mutant Comparison -- 8.4 Notes -- References -- Part III: Phenotypic Screening -- Chapter 9: Field Evaluation of Mutagenized Rice Material -- 9.1 Introduction -- 9.2 Materials -- 9.2.1 Plot Design -- 9.2.2 Field Preparation and Planting -- 9.2.3 Rice Culture -- 9.2.4 Field Observations and Trait Evaluation -- 9.3 Methods -- 9.3.1 Plot Design -- 9.3.2 Field Preparation and Planting -- 9.3.3 Rice Culture -- 9.3.4 Field Observations and Trait Evaluation -- 9.4 Notes -- References -- Chapter 10: Root Phenotyping Pipeline for Cereal Plants -- 10.1 Introduction -- 10.1.1 Issue of Root Phenotyping -- 10.1.2 Root Phenotyping of Cereal Plants -- 10.1.3 Proposed Root Phenotyping Pipeline -- 10.2 Materials -- 10.2.1 Design of a Plant Growth System -- 10.2.2 Root Scanning Setup -- 10.3 Methods -- 10.3.1 Preparation of Culture Media -- 10.3.2 Controlling the System and Monitoring the Medium Parameters -- 10.3.3 Experiment Preparation and Maintenance -- 10.3.4 Medium Exchange -- 10.3.5 Experiment Termination and Root System Cleaning -- 10.3.6 Root System Analysis Using WinRHIZO System -- 10.3.7 Root Image Analysis -- 10.4 Notes -- References -- Chapter 11: Breeding New Aromatic Rice with High Iron Using Gamma Radiation and Hybridization -- 11.1 Introduction -- 11.2 Materials -- 11.3 Methods -- 11.3.1 Preparing a Mutant Population.

11.3.2 Phenotypic Analysis of Aroma.