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Towards the Development of Long-Life Crops by Genetic Engineering of Ethylene Sensitivity
Ezura, Hiroshi The Korean Society of Plant Biotechnology 2000 식물생명공학회지 Vol.27 No.4
Food production is a major role of agriculture. It has been projected that the world population continues to increase by the middle of the 21st century, and the population growth results in raising a serious problem of food shortage. Thus we have to increase food as possible. A considerable amount of crops have been abandoned due to short-life after postharvest. Ethylene is a factor responsible for the postharvest loss in crops, especially horticultural crops. If we can reduce ethylene production or sensitivity by genetic engineering, we can develop, so called,“long-life crop”conferring long postharvest lives. During last two decades, intensive research for molecular dissection of ethylene biosynthesis has been carried out, and the researchers have succeeded in engineering ethylene productivity in some crops. On the other hand, after the successful isolation of Arabidopsis ethylene receptor gene ETR1, the homolog genes have been isolated in various plant species. Currently the characterization of these genes and alteration of ethylene sensitivity using the genes are in progress. This review summarizes current progress in the analysis of these genes, and discusses genetic engineering of ethylene sensitivity using these genes.
Rapid Tomato Breeding by Gene Editing Technologies
Hiroshi Ezura 한국원예학회 2021 한국원예학회 학술발표요지 Vol.2021 No.10
Gene editing technology is a new tool for rapid breeding including horticultural crops. We are trying to apply this technology for tomato improvements due to an availability of rich information of target genes. As a case study, I introduce an improvement of a nutritional trait, GABA (γ-aminobutyric acid) content, in tomato fruits. Here I would like to share our challenge and experience in development and implementation of the high GABA tomato. GABA is a non-proteinogenic amino acid with health-promoting functions for human. Although tomato fruits have a relatively high GABA content compared with other crops, the levels must be further increased to effectively confer the health-promoting functions such as lowering blood pressure. Glutamate decarboxylase (GAD) is a key enzyme in GABA biosynthesis in tomato; it has a C-terminal autoinhibitory domain that regulates enzymatic function, and deletion of this domain increases GAD activity. The tomato genome has five GAD genes, of which two are expressed during tomato fruit development. Then, to increase GABA content in tomato, we deleted the autoinhibitory domain of SlGAD3 using CRISPR/Cas9 technology. Introducing a stop codon before the autoinhibitory domain in the SlGAD3 increased GABA accumulation. We also evaluated the potential of the gene-edited tomato as a breeding material for hybrid tomatoes. Hybrid lines were produced by crossing the gene-edited tomato with a pure line cultivar. The hybrid lines showed high GABA accumulation in the fruits, which was sufficiently high for expecting health-promoting functions and had minimal effects on other fruit traits, suggesting that the high GABA is a dominant trait and that the gene-edited tomato would be useful as a parental line of hybrid cultivars. A university-lunched venture company has developed a commercial cultivar using the gene editing technology, and has started providing the high GABA tomato cultivar to home-gardener in May, 2021. In addition, I will introduce regulatory consideration and its recent progress in Japan.
Systematic Development of Tomato BioResources in Japan
Ariizumi, Tohru,Aoki, Koh,Ezura, Hiroshi Korean Society for Bioinformatics 2011 Interdisciplinary Bio Central (IBC) Vol.3 No.1
Recently, with the progress of genome sequencing, materials and information for research on tomato (Solanum lycopersicum) have been systematically organized. Tomato genomics tools including mutant collections, genome sequence information, full-length cDNA and metabolomic datasets have become available to the research community. In Japan, the National BioResource Project Tomato (NBRP Tomato) was launched in 2007, with aims to collect, propagate, maintain and distribute tomato bioresources to promote functional genomics studies in tomato. To this end, the dwarf variety Micro-Tom was chosen as a core genetic background, due to its many advantages as a model organism. In this project, a total of 12,000 mutagenized lines, consisting of 6000 EMS-mutagenized and 6000 gamma-ray irradiated M2 seeds, were produced, and the M3 offspring seeds derived from 2236 EMS-mutagenized M2 lines and 2700 gamma-ray irradiated M2 lines have been produced. Micro-Tom mutagenized lines in the M3 generation and monogenic Micro-Tom mutants are provided from NBRP tomato. Moreover, tomato cultivated varieties and its wild relatives, both of these are widely used for experimental study, are available. In addition to these bioresources, NBRP Tomato also provides 13,227 clones of full-length cDNA which represent individual transcripts non-redundantly. In this paper, we report the current status of NBRP Tomato and its future prospects.
Chikahiro Imashiro,Takashi Morikura,Motoaki Hayama,Atsushi Ezura,Jun Komotori,Shogo Miyata,Sakaguchi Katsuhisa,Shimizu Tatsuya 한국생물공학회 2023 Biotechnology and Bioprocess Engineering Vol.28 No.1
Various culture devices have been developed as fundamental technologies for facilitating bioengineering studies. Culture devices are designed to prepare specific culture environments. Thus, both macrostructures and surface micromorphology should be considered in the device design. Although fabricating devices with elaborate designs incurs high production costs, disposable materials are typically used for culture devices. However, some metallic materials are strong, stable, and biocompatible. Bioengineers have not applied these materials to culture devices because of the difficulty of processing. An emerging technology using three-dimensional (3D) printing has been developed, which can produce complex designs using metal. We demonstrate the applicability and potential of metal 3D printing for fabricating culture devices toward the development of the bioengineering discipline. As a specific example, we fabricated metallic culture devices where the environment of cultured tissues can be improved. One of the biggest factors determining the culture environment is active media supply. To attain active media supply to the tissue, devices having culture surfaces with mesh structures having holes far larger than cells were proposed. Cell sheets were cultured as tissue models, realizing tissue culture with such structures. The cultured tissue showed increased metabolism, indicating enhanced media supply owing to mesh surfaces. The biocompatibility of the 3D printed metal device was confirmed by viability assays on cultured cells, and reusability of the device was confirmed by mechanical and biochemical evaluations. We believe this study serves as a reference for using metallic 3D printing as an option for fabricating culture devices, which will promote bioengineering research.