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Mutation Breeding in Oil Palm: A Manual. Techniques in Plantation Science

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This is a practical guide to mutation breeding in oil palm, representing completely novel work supported by the Plant Breeding and Genetics Section of the Joint FAO/IAEA Division (Vienna, Austria). Oil palm is the top oil crop and the only major crop and only oil crop not to have been improved by plant mutation breeding. The manual is hands-on, providing step-by-step illustrated methods in mutation induction, mutation detection and mutant line development for oil palm improvement. Presenting sound practices based on scientific innovation and knowledge, this guide provides techniques integrated with expertise and is authored by practitioners actively engaged in oil palm seed production and breeding. Promoting green, eco-friendly agriculture, this book features coverage of: Radio-sensitivity testing; Challenges and opportunities for mutation breeding; Protocol for developing mutant generations for mutant screening; Services in irradiation treatments. The only available resource containing protocols and guidelines on how oil palm can be manipulated for mutation breeding, this book is essential reading for oil palm breeders, seed producers and plantation companies, oil palm traders, students and research institutes across the world. It provides a resource for training, a knowledge base for people new to oil palm and a reference guide for managers, to ensure best practices in maximising sustainability and production of this important crop.

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1 Introduction

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Introduction

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Abstract

A brief and general description of mutation breeding is provided. Mutation has been a successful strategy in producing over 3000 mutant cultivars in over 200 crop species worldwide. Oil palm is one of the few major crop species, and the only oil crop not to have been improved by mutation breeding.

However, pioneering work in mutation breeding in oil palm did take place in Ghana in the 1970s. This produced the first M1 population in oil palm and has only recently been progressed by developing M2 populations and in discovering mutants for crop improvement.

1.1  Brief History of Plant Mutation Breeding

The history of plant breeding spans centuries. The improvement of crop plants has been continuous since the domestication of species: initially by simple selection of naturally occurring forms; later by deliberate intervention. A landmark for plant breeding was the establishment of the laws of inheritance (Mendel, 1866), which transformed plant breeding from an art into a science. Since then, various plant breeding technologies have had significant impacts on plant breeding. These include: induced polyploidy; interspecific hybridization; chromosome engineering; alien gene introgression; doubled haploidy; F1 hybrids; transformation; genetic modification

 

2 Health and Safety Considerations and Guidelines

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Health and Safety Considerations and Guidelines

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Abstract

Mutation induction is the most hazardous part of mutation breeding. The application of physical or chemical mutagens as well as the general handling of mutant materials in mutation breeding has associated high health and safety risks. Poor practice can be hazardous, resulting in minor or serious personal injuries, and even fatal accidents. Therefore, the use of standard health and safety protocols as guides in mutation induction activities is an absolute necessity.

It is important to abide by the rules and regulations of mutation induction in order to avoid, or reduce to the barest minimum, injuries and accidents, as well as associated financial costs. With respect to chemical mutagens, these need specialist waste disposal systems after use. It is essential that mutation induction is carried out by specially trained personnel, and it is recommended that this is done in specialized institutes, or at least laboratories. International, regional and national service facilities are available (Chapter 6 of this manual). Certain rules and regulations constituting the operation guidelines may be generally applicable; however, depending on local requirements, some rules and regulations may be specific to a particular region or country. In certain jurisdictions, negligence in the strict adherence to health and safety protocols may incur sanctions such as fines, jail terms, or an embargo on field and laboratory research or commercial operations. This chapter highlights the standard operating protocols on health and safety issues relating to mutation induction for breeding in oil palm. Once mutation treatment has been carried out, subsequent steps

 

3 Radio-sensitivity Testing

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Radio-sensitivity Testing

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Abstract

Before physical mutagenesis can be applied for plant breeding purposes, the optimum irradiation dose needs to be determined. This is done using a radio-sensitivity test in which a range of dose treatments are applied to the target material. After irradiation treatments, the materials are grown out and recordings made for growth reduction (RD) and survival (LD). Normally, values of RD30–60 or LD30–60 are selected. Here, the target materials for radio-sensitivity testing are germinated seed.

3.1  Need for Radio-sensitivity Testing

Radio-sensitivity, or determination of the optimum dose of radiation, is a term describing a relative measure of the quantity of recognizable effects of a radiation exposure on the irradiated material (Owoseni et al., 2007). The plant breeder considers dose optimization as the first step in inducing a mutation breeding programme (Rohani et al., 2012). The optimal dose is one that produces sufficient mutation events so that there is a reasonable chance of the desired mutation occurring in the mutant population produced, but with a low background mutational load. Radio-sensitivity tests are performed with a wide range of doses to estimate the dose that produces effects normally between LD30 and LD50, and RD30 and RD50, when using a lethal parameter for the progeny and biomass parameter, respectively. These ranges have been observed to preserve the species integrity of the M1 (first mutant population) with the least possible unintended damage, but may be altered depending on a range of factors–for example, the breeding system of the species (Mba et al.,

 

4 Options for Mutation Breeding in Oil Palm

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Options for Mutation Breeding in

Oil Palm

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Abstract

Theoretical and practical options for mutation breeding in oil palm are discussed. In theory, haploids are considered the ideal targets, as the induced mutation can be fixed instantly on conversion to doubled haploids. However, haploid/doubled haploid technology is in its infancy in oil palm and therefore other practical targets need to be considered. The two obvious targets are pollen and seed, as these are produced in large numbers in breeding and commercial seed production. Schemes for pollen and seed irradiation and subsequent mutant population development are compared. The irradiation of germinated seed is currently considered to be the better approach in terms of convenience and time. A major constraint in mutation breeding in oil palm is the long life cycle. Oil palm has a long juvenile stage, and it takes 4–5 years from sowing a seed to getting seed of the next generation. Traditionally, mutant selection has relied on phenotypic selection, which can only take place in the second mutant generation (M2) due to the presence of physiological disorders and chimeras in the M1. However, now that the oil palm genome has been sequenced, it is feasible to select for mutants genotypically in the

 

5 Protocol for Developing Mutant Generations for Mutant Selection

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Protocol for Developing Mutant

Generations for Mutant Selection

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Abstract

A practical step-by-step protocol is presented for mutation induction and mutation detection in oil palm. Germinated seed is chosen as the target material for mutation induction, as this provides the quickest development of mutant populations (as discussed in Chapters 3 and 4 of this manual). The protocol adopts gamma irradiation as this is a proven effective mutagen for mutation breeding. Genotyping is deployed in the M1 to select for mutants in target genes. These selections, plus a random selection of M1 plants, are then advanced from nursery to field conditions to produce mature palms, which may be self-pollinated to produce the M2 generation. The M2 generation is subject to phenotypic screening at all stages in plant development, from seed, germination, seedling, juvenile to adult palms. A list of target genes and traits for mutation is given.

As discussed in Chapter 4 of this manual, there are two practical targets for mutation induction in oil palm: pollen and seed. The seed option is the more favoured, as it takes less time and involves only one pollination/seed production stage, thus saving time and labour. A step-by-step guide is provided in generating the M1 and M2 populations for mutant detection.

 

6 Services in Irradiation Treatments

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Services in Irradiation Treatments

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Abstract

Physical mutagens, particularly gamma and X-rays, are well established in plant mutation breeding. Although gamma irradiation has been predominant, it involves the use of radioactive isotopes and requires specialist facilities

(gamma cells, gamma houses and gamma fields). X-ray machines are more abundant and easier to use, and so are becoming more popular as they do not involve radioactive isotopes and therefore are not governed by the same stringent regulations as gamma emitters. A brief description and comparison of gamma and X-ray irradiation is given. Physical irradiation services are available at the international, regional and national levels. Other non-physical mutagens are discussed in relation to plant breeding and functional genomics.

6.1  Physical Mutagens

Physical mutagens include ultraviolet light (a non-ionizing radiation) and several types of ionizing radiation: gamma, X-ray, alpha and beta particles, ion beam, ion implantation, protons and fast neutrons. Of these, gamma and X-ray are the most commonly used in plant mutagenesis, although new methods, particularly in the use of ion beam and ion implantation, are being developed. Details of the main physical mutagens used in plant mutagenesis are presented in Shu et al. (2012a,b), who provide information on their physical characteristics, mode of action and how they may be utilized in plant breeding and genetics.

 

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