A5035962877- Transgenic Plants and Applications
- CRISPR and Genetic Engineering
- Plant tissue culture and regeneration
- Plant Virus Research Studies
- Biochemical Analysis and Sensing Techniques
- Plant Physiology and Cultivation Studies
- Postharvest Quality and Shelf Life Management
- Genetically Modified Organisms Research
- Plant Reproductive Biology
- Glycosylation and Glycoproteins Research
- Plant Molecular Biology Research
- Plant Genetic and Mutation Studies
- Insect Resistance and Genetics
- Metabolomics and Mass Spectrometry Studies
- Chromosomal and Genetic Variations
- Plant biochemistry and biosynthesis
- Analytical Chemistry and Chromatography
- RNA and protein synthesis mechanisms
University of Tsukuba
2010-2022
Innovation Research Center
2021
Okayama University
2003
The tomato is an excellent model for studies of plants bearing berry-type fruits and experimental the Solanaceae family due to its conserved genetic organization. In this study, a comprehensive mutant population was generated in background Micro-Tom, dwarf, rapid-growth variety. previous studies, including 8,598 6,422 M2 mutagenized lines produced by ethylmethane sulfonate (EMS) mutagenesis γ-ray irradiation, study developed investigated these alteration visible phenotypes. A total 9,183...
In order to investigate the physiological role of ethylene in initiation and subsequent progression softening, pear fruit were treated with propylene, an analogue or 1-methylcyclopropene (1-MCP), a gaseous inhibitor action at preclimacteric ripening stages. The propylene treatment pre-ripe stage stimulated production flesh softening while 1-MCP same markedly retarded ripening-related events. Moreover, after suppressed production. These results clearly indicate that is not merely by-product,...
As metabolomics can provide a biochemical snapshot of an organism's phenotype it is promising approach for charting the unintended effects genetic modification. A critical obstacle this application inherently limited metabolomic coverage any single analytical platform. We propose using multiple platforms direct acquisition interpretable data set estimable chemical diversity. example, we report our multi-platform that assesses substantial equivalence tomatoes over-expressing taste-modifying...
A transgenic tomato line (56B, "Moneymaker") that expresses the miraculin gene driven by CaMV 35S promoter was crossed with a dwarf ("Micro-Tom") for molecular breeding of cultivars are suitable production in closed cultivation system. Plant size, accumulation, and self-pruning growth were used as selection indicators F2 plants. Two lines chosen further analysis, bred to F6 or F7 generation cultivated In 56B two lines, concentrations pericarp 140, 367, 343 microg/g FW, respectively. We also...
High-level accumulation of the target recombinant protein is a significant issue in heterologous expression using transgenic plants. Miraculin, taste-modifying protein, was accumulated tomatoes an cassette which miraculin gene expressed by cauliflower mosaic virus (CaMV) 35S promoter and heat shock (HSP) terminator (MIR-HSP). The HSP derived from 18.2 Arabidopsis thaliana. Using this HSP-containing cassette, concentration T0 tomato lines 1.4–13.9% total soluble (TSP), that T1 line homozygous...
One of the ultimate goals plant science is to test a hypothesis obtained by basic and apply it agriculture industry. A factory one ideal systems for this trial. Environmental factors affect both yield accumulation recombinant proteins industrial applications within transgenic plants. However, there have been few reports studying productivity protein in closed cultivation called factories. To investigate effects photosynthetic photon flux (PPF) on tomato fruit miraculin, taste-modifying...
We previously developed a transgenic tomato that expresses the miraculin gene using constitutive promoter. In this study, we profiled developmental and spatial accumulation of protein mRNA in fruits. Miraculin expression was almost constant up to orange stage, then increased at red stage. The accumulated gradually during fruit development reached its highest level overripe At stage fruit, exocarp, similar other tissues: mesocarp, dissepiment, upper placenta, lower placenta jelly. Moreover,...
Miraculin is a taste-modifying protein that can be isolated from miracle fruit (Richadella dulcifica), shrub native to West Africa. It able turn sour taste into sweet taste. The commercial exploitation of this sweetness-modifying underway, and fast efficient purification method extract the needed. We succeeded in purifying miraculin single-step using immobilized metal-affinity chromatography (IMAC). purified exhibited high purity (>95%) reverse-phase high-performance liquid chromatography....
Gene dosage and genetic background are factors that influence transgene expression in transgenic plants. In our previous studies, we produced tomato plants accumulate miraculin, a taste-modifying protein, genetically stable manner. To elucidate the effects of gene on miraculin accumulation fruits, generated hybrid lines between homozygous line 56B (background cultivar ‘Moneymaker’) pure cultivars ‘Micro-Tom,’ ‘Moneymaker,’ ‘Ailsa Craig,’ ‘M82,’ ‘Rutgers’ ‘Aichi-first’ analyzed them for mRNA...
The sweet taste-modifying protein miraculin has been successfully expressed in the tomato, a high-yield commercial plant (Sun et al. 2007). Previously proposed methods of purifying untagged from transgenic tomato fruit are laborious and time consuming do not allow separation undenatured denatured miraculin. Therefore, it is necessary to develop an efficient method fruit. In this study, we have demonstrated that combined use nickel-immobilized affinity chromatography ion-exchange readily...
Abstract High target protein concentrations in source materials are important for achieving high purification efficiency. We sought to produce the taste-modifying protein, miraculin, transgenic tomato fruits and then extract purify this protein. In order improve efficiency of recombinant miraculin from tomatoes, we tested a salt-stress cultivation technique aimed at increasing concentration tomatoes. Two lines 56B 5B, were grown under conditions. There was higher content per gram fresh...
Miraculin is a promising protein with taste-modifying properties. Focusing on the unique function and potential of miraculin, recombinant miraculin production has been explored use heterologous expression systems, but activities miraculins were much lower than those native probably due to difference in post-translational modification, especially N-glycosylation. For practical therefore, differences between N-glycan compared that should be minimized. Here, establish platform for functional...