Application of in vivo and in vitro mutation techniques for crop improvement

Summary Conventional mutation techniques have often been used to improve yield, quality, disease and pest resistance in crops, or to increase the attractiveness of flowers and ornamental plants. More than 1700 mutant varieties involving 154 plant species have been officially released. In some economically important crops, e.g. barley, durum wheat and cotton, mutant varieties occupy the majority of cultivated areas in many countries. Mutation techniques have become one of the major tools in the breeding of ornamentals such as alstroemeria, begonia, chrysanthemum, carnation, dahlia and streptocarpus. The use of in vitro techniques such as anther culture, shoot organogenesis, somatic embryogenesis and protoplast fusion can overcome some of the limitations in the application of mutation techniques in both seed and vegetatively propagated crops. In vitro culture in combination with induced mutations can speed up breeding programmes, from the generation of variability, through selection, to multiplication of the desired genotypes. The expression of induced mutations in the pure homozygote obtained through microspore, anther or ovary culture, can enhance the rapid recovery of the desired traits. In some vegetatively propagated species, mutations in combination with in vitro culture technique, may be the only method of improving an existing cultivar. Currently, many molecular studies rely on the induction and identification of mutants in model species for construction and subsequent saturation of genetic maps, understanding of developmental genetics and elucidation of biochemical pathways. Once identified and isolated, the genes that encode agronomically-important features can be either introduced directly into crop plants or used as probes to search for similar genes in crop species. It seems most likely that the recent developments based on these technologies will soon provide improved methods for selection of desired mutants.     

Tissue culture-derived variation in crop improvement

Tissue culture generates a wide range of genetic variation in plant species which can be incorporated in plant breeding programmes. By in vitro selection, mutants with useful agronomic traits, e.g. salt or drought tolerance or disease resistance, can be isolated in a short duration. The successful use of somaclonal variation is very much dependent on its genetic stability in the subsequent generations for which molecular markers such as RAPDs, AFLPs, SSRs and others can be helpful. The potential of somaclonal variation has yet to be fully exploited by breeders, even though a few cultivars have been developed in crops such as Brassica juncea, rice and others. This revised version was published online in July 2006 with corrections to the Cover Date.     

Challenges in wide implementation of genome editing for crop improvement

Genome editing technologies using customizable sequence specific nucleases (SSNs) enables precise genome modification in plants, which has revolutionized functional genomics research as well as crop improvement. Applicability of SSN-mediated genome editing ranges from targeted gene knockout and single base modification to multiple gene stacking into desired genomic sites. However, there are still considerable challenges in implementing genome editing technologies for practical crop improvement. Here, we briefly discuss the technological challenges, especially those associated with the delivery of SSNs and with homologous recombination-mediated genome editing, and address some promising solutions to overcome the issues.     

Somaclonal variation as a tool for crop improvement

Summary Somaclonal variation is a tool that can be used by plant breeders. The review examines where this tool can be applied most effectively and the factors that limit or improve its chances of success. The main factors that influence the variation generated from tissue culture are (1) the degree of departure from organised growth, (2) the genotype, (3) growth regulators and (4) tissue source. Despite an increasing understanding of how these factors work it is still not possible to predict the outcome of a somaclonal breeding programme. New varieties have been produced by somaclonal variation, but in a large number of cases improved variants have not been selected because (1) the variation was all negative, (2) positive changes were also altered in negative ways, (3) the changes were not novel, or (4) the changes were not stable after selfing or crossing. Somaclonal variation is cheaper than other methods of genetic manipulation. At the present time, it is also more universally applicable and does not require containment procedures. It has been most successful in crops with limited genetic systems and/or narrow genetic bases, where it can provide a rapid source of variability for crop improvement.     

In vitro adventitious bud techniques for vegetative propagation and mutation breeding of potato (Solanum tuberosum L.). II. Significance for mutation breeding

Summary It was investigated what potentialities for mutation breeding of potato are offered by using adventitious sprouts that arise in vitro from leaf explants (rachis, petiole, leaflet-disc) after X-irradiation. Mutation frequency and chimerism were studied in subterranean and aerial parts in three vegetative generations (vM₁, vM₂, vM₃). Plants obtained from irradiated series produced a very high mutation frequency, a wide mutation spectrum and a very low rate of chimerism. Mutations were observed also in control series, especially in plants derived from the rachis and petiole explant group.     

Relevance of sexual polyploidization for crop improvement – A review

Colchicine induced polyploids have not directly contributed for crop improvement in the past. On the other hand, the so-called natural polyploids, derived from the functioning of numerically unreduced(2n) gametes have been shown to be more relevant for crop improvement in many cases. Different types of cytological abnormalities during meiosis can give rise to 2n gametes and the genetic composition of these gametes is variable. Depending on the type meiotic abnormalities, various types of 2ngametes, such as first division restitution(FDR), second division restitution (SDR),indeterminate meiotic restitution (IMR) and post meiotic restitution (PMR) gametes,among others, have been described in recent years. For the improvement of autopolyploids such as potato, alfalfa,Vaccinium spp., and some of the fodder grasses, FDR gametes have been proved to be highly useful. However, the use of 2n gametes for the improvement of allopolyploid crops has received much less attention so far. Some of the investigations on allopolyploids, derived from Festuca-Lolium, Alstroemeria and Lilium species hybrids, have revealed that 2ngametes can be most useful for the introgression of alien genes and chromosomes into cultivars. An important feature of using sexual polyploidization in the case of allopolyploids is that introgression can be achieved through recombination due to genetic crossing-over between alien chromosomes as well as addition of alien chromosomes, which is extremely difficult or impossible to achieve in the case of colchicine induced allopolyploids. Because of the recent developments in the field of plant molecular biology, methods have become available for the analysis of 2ngametes and sexual polyploid progenies more accurately and to develop systematic breeding approaches. The methods include DNA in situ hybridization (GISH and FISH)and molecular mapping (AFLP, RFLP, RAPDs).In addition to providing basic information on the genetic and genome composition of the polyploid progenies, these methods can be potentially useful for a more efficient creation of desirable breeding material and cultivars. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic improvement for root growth angle to enhance crop production

The root system is an essential organ for taking up water and nutrients and anchoring shoots to the ground. On the other hand, the root system has rarely been regarded as breeding target, possibly because it is more laborious and time-consuming to evaluate roots (which require excavation) in a large number of plants than aboveground tissues. The root growth angle (RGA), which determines the direction of root elongation in the soil, affects the area in which roots capture water and nutrients. In this review, we describe the significance of RGA as a potential trait to improve crop production, and the physiological and molecular mechanisms that regulate RGA. We discuss the prospects for breeding to improve RGA based on current knowledge of quantitative trait loci for RGA in rice.     

Mutation breeding of Chrysanthemum morifolium Ram. using in vivo and in vitro adventitious bud techniques

Summary During experiments, which are being carried out to study the factors which control the process of adventitious bud formation in vivo on detached leaves of Chrysanthemum morifolium Ram, adventitious shoots were produced from leaves, irradiated with 500 rad of X-rays. The most important but disadvantageous result was that the majority of the adventitious shoots proved to be of a chimeral nature and obviously developed from more than one cell.An in vitro adventitious bud technique was developed using different types of explants. Pedicel segments regenerated the highest number of adventitious shoots and, moreover, they developed faster as compared to explants of young flower heads or leaves. The mutants produced by irradiating the various explants were almost exclusively of a solid (non-chimeral) nature. In addition, histological observations suggest that single epidermal cells are involved in the initiation of the adventitious shoot apices.The optimum dose for mutant production is approximately 800 rad X-rays. Rather often, more than one phenotypically identical mutant was found, which was always derived from the same explant. They could for instance originate from a multi-apical meristem formed by a single mutated cell.     

Collaboration of farmers and breeders: Participatory crop improvement in perspective

Participatory Crop Improvement (PCI) has developed over the past decade as an alternative and complementary breeding approach to Formal Crop Improvement (FCI). In that context, PCI principally aims at more effectively addressing the needs of farmers in marginal areas in developing countries. This paper describes the rationale behind the emerging of the PCI-concept, the first experiences, and its place in a development-context. The relation with in situ conservation of plant genetic resources is briefly described. The paper uses the distinction in PCI between PVS (Participatory Varietal Selection, i.e. participatory selection among varieties and advanced materials) and PPB (Participatory Plant Breeding, i.e. selection within segregating materials). While successful experiences of PVS are reported, the potentials of PPB are still to be explored. Among other issues, this article pays attention to Genotype x Environment interaction: while G × E interaction is recognised as an important issue in plant breeding and a justification for PCI, the implications for the design of selection systems with farmer participation, and, eventually the potential of PPB has only been analysed to a limited extend. Questions in relation to materials, breeding strategies and selection procedures to achieve sufficient progress in the different crops and environments are identified; these will however remain unanswered until more experiences from the field are available. This revised version was published online in August 2006 with corrections to the Cover Date.     

农作物转基因技术研究进展及存在问题

就转基因应用所面临的主要技术问题--外源基因的逃逸、失活和沉默进行了分析,解读了该现象发生的机理,并指出可以在操作过程中通过时空隔离、避免基因间的同源性、避免重复序列的出现、消除DNA甲基化的影响、使用MAR以及使用诱导型启动子等方法,对上述现象加以控制.最后,对转基因技术的应用前景作了积极的展望.     

The potential applications of site-directed mutagenesis for crop improvement: a review

Journal of Crop Science and Biotechnology - The search for technologies for crop improvement has been a continuous practice to address the food insecurity to the growing human population with an...     

Current status and future strategies of in vitro culture techniques for genetic improvement of mungbean (Vigna radiata (L.) Wilczek)

Summary Mungbean is an important source of vegetable protein for the growing population in many developing countries of South East Asia. Its production is limited due to its susceptibility to diseases and insect pests besides many other undesirable agronomic traits. Strategies for increasing and stabilising its production have been to develop varieties resistant to diseases, pests and with other desirable agronomic traits. Genetic improvement of this crop by classical breeding has met with limited success due to the lack of sufficient and satisfactory level of genetic variability within germplasm. Recent advances in biotechnology have offered the opportunity to develop new germplasms. The development of such technologies largely depends on efficient regeneration of sexually mature plants from organs, tissues and protoplasts. An overview of plant regeneration by direct or indirect organogenesis and embryogenesis is presented. The use of in vitro and molecular techniques such as somaclonal variation, screening for various desirable traits, interspecific crossing and genetic transformation to supplement conventional breeding, for genetic improvement of this crop is described. The advantages and limitations of these techniques along with directions for future research are discussed.     

转基因技术在玉米种质改良中的应用

简述了玉米转基因工程中遗传转化的受体系统、目的基因和转基因技术的研究进展.分析了转基因技术在改良玉米品质、抗虫、抗病、抗除草剂、耐盐、抗旱、抗寒性、雄性不育、抗真菌和抗病毒病基因等方面取得的成效.     

Participatory plant breeding research: Opportunities and challenges for the international crop improvement system

This paper describes the current state of international plant breeding research and explains why the centralized global approach to germplasm improvement that was so successful in the past is today being transformed by the incorporation of decentralized local breeding methods designed to better incorporate the perspective of end users into the varietal development process. It describes international breeding efforts for major crops and identifies factors that have contributed to the success of the international breeding system; discusses shortcomings of the global approach to plant breeding and explains why future successes will depend critically on researchers' ability to incorporate the knowledge and preferences of technology users; reviews a number of farmer participatory research methods that are currently being tested by plant breeding programs throughout the developing world; describes synergies that can potentially be achieved by linking centralized global and decentralized local breeding models; and discusses technical, economic, and institutional challenges that will have to be overcome to integrate end user-based participatory approaches into the international plant breeding system. This revised version was published online in August 2006 with corrections to the Cover Date.     

In vitro techniques for genomic alteration in rice plants

Because of the explosive increase in world population, a sufficient food supply must be achieved by varietal improvement in the major cereal crops including rice. It is expected that new in vitro techniques incombination with conventional breeding methods may effectively raise the yield potential. On the other hand, there are many environmental problems to be solved world-wide such as, global warming, environmental pollution, ecological destruction, reduction in water supplies and so on. Therefore, it is necessary to rapidly develop new varieties for the future combining of higher yield potential with excellent grain quality, and resistance to both biotic and abiotic stresses for the promotion of sustainable agriculture. Although many efforts have been made to introduce useful traits from wild species to cultivated rice via hybridization, it is still difficult to overcome breeding barriers such as cross incompatibility and hybrid sterility and inviability in practical breeding. Now in vitro techniques are going to make it possible to use genetic manipulation and cell culture and fusion techniques to speed up the breeding process. For sustainable agriculture, it is important to utilize the useful genes from alien species. For this purpose, asymmetric protoplast fusions have already been used successfully to transfer disease resistance in Brassica napus. In this experiment, a high level of resistance to the rice blast disease was transferred from wild species through asymmetric fusions. It is also noted that manipulation of cytoplasmic genomes is possible through asymmetric fusions as shown in the induction of new cytoplasmic male sterility (CMS). This revised version was published online in August 2006 with corrections to the Cover Date.