Sakurai T.,RIKEN |
Kondou Y.,RIKEN |
Kondou Y.,Kanto Gakuin University |
Akiyama K.,RIKEN |
And 28 more authors.
Plant and Cell Physiology | Year: 2011
Identification of gene function is important not only for basic research but also for applied science, especially with regard to improvements in crop production. For rapid and efficient elucidation of useful traits, we developed a system named FOX hunting (Full-length cDNA Over-eXpressor gene hunting) using full-length cDNAs (fl-cDNAs). A heterologous expression approach provides a solution for the high-throughput characterization of gene functions in agricultural plant species. Since fl-cDNAs contain all the information of functional mRNAs and proteins, we introduced rice fl-cDNAs into Arabidopsis plants for systematic gain-of-function mutation. We generated >30,000 independent Arabidopsis transgenic lines expressing rice fl-cDNAs (rice FOX Arabidopsis mutant lines). These rice FOX Arabidopsis lines were screened systematically for various criteria such as morphology, photosynthesis, UV resistance, element composition, plant hormone profile, metabolite profile/fingerprinting, bacterial resistance, and heat and salt tolerance. The information obtained from these screenings was compiled into a database named 'RiceFOX'. This database contains around 18,000 records of rice FOX Arabidopsis lines and allows users to search against all the observed results, ranging from morphological to invisible traits. The number of searchable items is approximately 100; moreover, the rice FOX Arabidopsis lines can be searched by rice and Arabidopsis gene/protein identifiers, sequence similarity to the introduced rice fl-cDNA and traits. The RiceFOX database is available at http://ricefox.psc.riken.jp/. © 2011 The Author.
Sasaki K.,Japan National Agriculture and Food Research Organization |
Kato K.,Nara Institute of Science and Technology |
Mishima H.,NEC Solution Innovators Ltd |
Furuichi M.,NEC Solution Innovators Ltd |
And 5 more authors.
Plant Biotechnology | Year: 2014
Florescent proteins have been popularly used for studying genes and proteins of interest in various experiments at a cellular level, such as the analysis of intracellular localization and protein–protein interaction. However, the strength of fluorescence was insufficient for macro level observations of tissues or of the whole plant, and the fluorescent flowers that have been generated so far needed high-sensitive imaging equipment for the observation. Here we generated fluorescent Torenia flowers by the combined use of a high-performance fluorescent protein and the latest protein expression technologies, leading to the production of fluorescent proteins that can be easily and clearly observed. A coding sequence of a yellowish green fluorescent protein from the marine plankton Chiridius poppei (CpYGFP) was fused to the optimized sequences of the heat shock protein terminator and the 5′-untranslated region of the alcohol dehydrogenase gene of Arabidopsis to gain massive accumulation of the fluorescent protein. Strong fluorescence of CpYGFP was apparent in every part of the transgenic plant under the simple combination of a blue LED for excitation and an orange colored transparent acrylic filter for emission, while faint autofluorescence remained in the wild-type plants. By evaluating the combination of excitation wavelengths (excitation and emission filters) we were able to eliminate this undesired fluorescence. The fluorescent flowers could be used for ornamental purposes as well as for the analysis of fluorescent transgenic plants spatiotemporally in a nondestructive manner. © 2014 The Japanese Society for Plant Cell and Molecular Biology.
Zhang H.,University of Tokyo |
Liang C.,University of Tokyo |
Aoki N.,University of Tokyo |
Kawai K.,Kumiai Chemical Industry Co. |
And 2 more authors.
Plant Production Science | Year: 2016
We report the effect of the introduction of a fungal glutamate dehydrogenase gene (gdhA) into forage rice (cv. Momiroman) by analyzing the transgenic rice plants in terms of growth, source function, and nitrogen contents. NADP (H)-dependent glutamate dehydrogenase activities in the gdhA-transgenic lines were markedly higher than those in non-transgenic control plants. Plant growth analysis at the seedling stage revealed that the leaf area and shoot and root dry weights of the high gdhA-expressors were higher than those of control plants under both high (high N) and low nitrogen (low N) conditions. These results suggested that the source ability was enhanced by the gdhA introduction. This was supported by the fact that the net photosynthesis rate at the heading stage was also higher in transgenic than in control leaves. Furthermore, under both high and low N conditions, the nitrogen contents in the shoots and roots, at seedling and grain-harvest stages, were significantly higher in high gdhA-expressors than in control plants, indicating that nitrogen uptake was higher in transgenic than in control plants. At the harvest stage, the high gdhA-expressors exhibited greater panicle and spikelet numbers per plant compared with control plants, resulting in higher grain weight, under the high N conditions. In addition, gdhA expression in forage rice significantly enhanced their tolerance to salt stress compared to control plants. The present study showed that the introduction of a fungal gdhA into forage rice could lead to higher source ability, better growth and higher grain weight by enhancing nitrogen uptake efficiency. © 2016 The Author(s). Published by Taylor & Francis.
Hiwasa-Tanase K.,University of Tsukuba |
Kuroda H.,Inplanta Innovations Inc. |
Hirai T.,University of Tsukuba |
Aoki K.,Osaka Prefecture University |
And 2 more authors.
Plant Cell Reports | Year: 2012
Fruit-specific promoters have been used as genetic engineering tools for studies on molecular mechanism of fruit development and advance in fruit quality and additional value by increasing functional component. Especially fruit-ripening specific promoters have been well utilized and studied in tomato; however, few studies have reported the development of promoters that act at fruit developing stages such as immature green and mature green periods. In this study, we report novel promoters for gene expression during the green to ripening stages of tomato fruit development. Genes specifically expressed at tomato fruit were selected using microarray data. Subsequent to confirmation of the expression of the selected 12 genes, upstream DNA fragments of the genes LA22CD07, Les. 3122.2. A1_a_at and LesAffx. 6852.1. S1_at which specifically expressed at fruit were isolated from tomato genomic DNA as promoter regions. Isolated promoter regions were fused with the GUS gene and the resultant constructs were introduced into tomato by agrobacterium-mediated transformation for evaluation of promoter activity in tomato fruit. The two promoters of LA22CD07, and LesAffx. 6852.1. S1_at showed strong activity in the fruit, weak activity in the flower and undetectable activity in other tissues. Unlike well-known fruit-ripening specific promoters, such as the E8 promoter, these promoters exhibited strong activity in green fruit in addition to red-ripening fruit, indicating that the promoters are suitable for transgene expression during green to ripening stages of tomato fruit development. Key message Novel fruit-specific promoters have been identified and are suitable for transgene expression during green to ripening stages of tomato fruit development. © 2012 Springer-Verlag.
Kato K.,University of Tsukuba |
Yoshida R.,University of Tsukuba |
Kikuzaki A.,Inplanta Innovations Inc. |
Hirai T.,University of Tsukuba |
And 5 more authors.
Journal of Agricultural and Food Chemistry | Year: 2010
A transgenic tomato line (56B, "Moneymaker") that expresses the miraculin gene driven by the CaMV 35S promoter was crossed with a dwarf tomato ("Micro-Tom") for the molecular breeding of cultivars that are suitable for miraculin production in a closed cultivation system. Plant size, miraculin accumulation, and self-pruning growth were used as selection indicators for F2 plants. Two lines were chosen for further analysis, bred to the F6 or F7 generation and cultivated in a closed cultivation system. In 56B and the two crossed lines, the concentrations of miraculin in the pericarp were 140, 367, and 343 μg/g FW, respectively. We also estimated that 26.2, 73.6, and 45.9 kg FW/m2 of tomatoes and 2.2, 16.6, and 9.8 mg/m2 of miraculin in the pericarp, respectively, could be harvested per year. These two crossed lines will be useful for the mass production of miraculin, especially in a closed cultivation system. © 2010 American Chemical Society.
Inplanta Innovations Inc., University of Tsukuba and Kazusa DNA Research Institute | Date: 2011-03-30
The present invention relates to a fruit-specific promoter, which is suitable for the expression in a broader range of developmental stages of a fruit. Provided is a fruit-specific promoter DNA, which consists of a nucleotide sequence having 85% or more identity with the nucleotide sequence as shown in SEQ ID NO: 1 or 2 and has promoter activity in mature-green fruits.