著者

Azusa Ono Kyoko Hiwasa-Tanase Satoko Nonaka Hiroshi Ezura

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.38, no.1, pp.161-165, 2021-03-25 (Released:2021-03-25)

参考文献数

21

被引用文献数

2

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The taste-modifying protein miraculin (MIR) has received increasing interest as a new low-calorie sweetener. In our previous study using the tomato variety ‘Micro-Tom,’ it was shown that in transgenic tomatoes in which MIR was expressed by using the cauliflower mosaic virus 35S promoter (p35S) and a heat shock protein terminator (tHSP) cassette (p35S-MIR-tHSP), higher levels of miraculin accumulated than when MIR was driven by the nopaline synthase terminator (tNOS) cassette (p35S-MIR-tNOS). ‘Micro-Tom’ is a dwarf tomato used for research and shows a low yield. To achieve high productivity of MIR, it is essential to improve the MIR accumulation potential by using high-yielding cultivars. In this study, we evaluate whether the high MIR accumulation trait mediated by the tHSP appears even when fruit size increases. A line in which the p35S-MIR-tHSP cassette was introduced into a high-yielding variety was bred by backcrossing. The line homozygous for MIR showed higher accumulation of MIR than the heterozygous line. Despite large differences in fruit size, the MIR level in the backcross line was similar to that in the p35S-MIR-tHSP line (background ‘Micro-Tom’). It was approximately 3.1 times and 4.0 times higher than those in miracle fruits and the p35S-MIR-tNOS tomato line 5B (‘Moneymaker’ background, which exhibits the highest miraculin productivity achieved thus far), respectively. These results demonstrate that the high MIR accumulation trait mediated by the tHSP appears even when fruit size is increased.

著者

Kyoko Hiwasa-Tanase Tsubasa Yano Tatsuya Kon Teruhiko Terakawa Hiroshi Ezura

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.38, no.4, pp.421-431, 2021-12-25 (Released:2021-12-25)

参考文献数

48

被引用文献数

1 3

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The commercial use of genetically modified (GM) crops requires prior assessment of the risks to the environment when these crops are grown in the field or distributed. Assessments protocols vary across countries and GM crop events, but there is a common need to assess environmental biosafety. In this study, we conducted an environmental risk assessment in a confined field of GM tomato plants that can produce miraculin, a taste-altering protein that causes sour tastes to be perceived as sweet, for practical use in Japan. The evaluation was conducted for 1) competitiveness (the ability to compete with wild plants for nutrients, sunlight, and growing areas and prevent their growth) and 2) the production of toxic substances (the ability to produce substances that interfere with the habitat and growth of wild plants, animals, and microorganisms). Investigations of plant morphology and growth characteristics as well as tolerance to low temperature during early growth and overwintering for assessment endpoints related to competitiveness showed no biologically meaningful difference between GM tomato and non-GM tomato. In addition, harmful substances in plant residues and root secretions were assessed by the plow-in method, succeeding crop test and soil microflora tests, and it was determined that GM tomato does not exhibit an increase in harmful substances. Based on these results, it was concluded that GM miraculin-accumulating tomato is comparable to conventional tomato and is unlikely to have unintended adverse effects in the natural environment of Japan.

著者

Toshifumi Nagata Fabien Lombardo Hiroshi Ezura

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.38, no.3, pp.387-390, 2021-09-25 (Released:2021-09-25)

参考文献数

14

被引用文献数

1

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The HAWAIIAN SKIRT (HWS) gene was originally described in Arabidopsis for the characteristic fusion of sepals in the mutant. A tomato line mutated in the putative ortholog gene was isolated in a previous study. The tomato hws-1 mutant showed facultative parthenocarpy and produced fruits with elevated Brix, revealing the gene as a hopeful resource for crop improvement. To confirm the orthology relationship between the Arabidopsis and tomato HWS genes, the hws-1 mutant was complemented with either the tomato wild-type genomic fragment or the Arabidopsis sequence of the gene. In both complementation experiments, defective phenotypes of hws-1 are rescued, albeit to different extents. Recovery of these phenotypes, which include parthenocarpic fruit production, increased Brix, loss of leaflet serration, alteration of bud and petal shape, firmly establishes SlHWS as an ortholog of the originally described HWS in Arabidopsis. This work indicates that the function of HWS is likely to be conserved in a wide range of plant species.

著者

Shohei Nosaki Ken Hoshikawa Hiroshi Ezura Kenji Miura

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.38, no.3, pp.297-304, 2021-09-25 (Released:2021-09-25)

参考文献数

60

被引用文献数

26

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The production of recombinant proteins is important in academic research to identify protein functions. Moreover, recombinant enzymes are used in the food and chemical industries, and high-quality proteins are required for diagnostic, therapeutic, and pharmaceutical applications. Though many recombinant proteins are produced by microbial or mammalian cell-based expression systems, plants have been promoted as alternative, cost-effective, scalable, safe, and sustainable expression systems. The development and improvement of transient expression systems have significantly reduced the period of protein production and increased the yield of recombinant proteins in plants. In this review, we consider the importance of plant-based expression systems for recombinant protein production and as genetic engineering tools.

著者

Shohei Nosaki Ken Hoshikawa Hiroshi Ezura Kenji Miura

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

pp.21.0610a, (Released:2021-09-18)

参考文献数

60

被引用文献数

26

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The production of recombinant proteins is important in academic research to identify protein functions. Moreover, recombinant enzymes are used in the food and chemical industries, and high-quality proteins are required for diagnostic, therapeutic, and pharmaceutical applications. Though many recombinant proteins are produced by microbial or mammalian cell-based expression systems, plants have been promoted as alternative, cost-effective, scalable, safe, and sustainable expression systems. The development and improvement of transient expression systems have significantly reduced the period of protein production and increased the yield of recombinant proteins in plants. In this review, we consider the importance of plant-based expression systems for recombinant protein production and as genetic engineering tools.

著者

Suzuka Yamagishi Koji Nakagawa Kazutomo Baba Hiroaki Kawamoto Yoshiyuki Sankai Naomichi Fujiuchi Hiroshi Ezura Naoya Fukuda

出版者

The Society of Eco-Engineering

雑誌

Eco-Engineering (ISSN:13470485)

巻号頁・発行日

vol.33, no.3, pp.79-85, 2021-07-31 (Released:2021-07-31)

参考文献数

29

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Near-infrared (NIR) spectroscopy has been used to predict fruit quality. However, thereʼs few previous studies which use small and lightweight devices and they didnʼt construct models for each cultivar although each tomato cultivar has its own characteristics such as color and size. In this study, models to nondestructively determine fruit quality of tomato cultivars A (orange-colored cherry tomato), B (red-colored cherry tomato), C (red-colored cocktail tomato), and D (orange-colored cocktail tomato) were constructed using an inexpensive, portable near-infrared (NIR) spectroscopic device (spectral response range: 590-1,050 nm). We irradiated the tomato fruits and analyzed the spectra of the scattered light. The spectral data were preprocessed using four different methods (min-max normalization, 1st differential, 2nd differential and combination of normalization and 1st differential). The soluble solid content (SSC) and citric acid content (CAC) of the fruits were determined by destructive methods. Partial least-squares regression was used to construct models for predicting the SSC and CAC from the preprocessed spectra. The best preprocessing method differed by cultivar. For the best and worst models of SCC, the R2 values were 0.80 and -0.1, respectively. Model performance may differ according to the color of the pericarp. Also, selection of an appropriate preprocessing method is important when using small spectroscopic devices to measure tomato fruit quality.

著者

Hiroyuki Kajiura Kyoko Hiwasa-Tanase Hiroshi Ezura Kazuhito Fujiyama

出版者

Japanese Society for Plant Cell and Molecular Biology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.35, no.4, pp.375-379, 2018-12-25 (Released:2018-12-31)

参考文献数

17

被引用文献数

1 4

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Miraculin is a promising protein with taste-modifying properties. Focusing on the unique function and potential of miraculin, recombinant miraculin production has been explored with the use of heterologous expression systems, but the activities of recombinant miraculins were much lower than those of native miraculin, probably due to the difference in post-translational modification, especially N-glycosylation. For practical use therefore, the differences between N-glycan of recombinant miraculin compared to that of native miraculin should be minimized. Here, to establish the platform for functional miraculin production, we expressed miraculin in tomato plants with the same taste-modifying activity as native miraculin purified from miracle fruit, and we compared the N-glycan structures with those of native miraculin. Our N-glycan structural analysis using purified miraculin, followed by hydrazynolysis, 2-pyridylamine (PA)-labeling, high-performance liquid chromatography, and a liquid chromatography tandem-mass spectrometry analysis revealed that both the native and recombinant miraculins carried an M3 structure as a predominant structure and that most of the N-glycan structures on the miraculins were pauci-mannosidic structures with a smaller amount of plant-specific α1,3-fucosylated and/or β1,2-xylosylated N-glycans and without a Lewis a epitope. These results indicate that the N-glycoform of native miraculin from miracle fruit and recombinant miraculin expressed in tomato plants are almost identical to each other with similar ratios and that, therefore, plant-specific N-glycans are essential for showing the full taste-modifying activity of miraculin.

著者

Moalla Rached Baldet Pierre Gibon Yves Chiaki Matsukura Toru Ariizumi Hiroshi Ezura Naoya Fukuda

出版者

The Japanese Society for Horticultural Science

雑誌

The Horticulture Journal (ISSN:21890102)

巻号頁・発行日

pp.OKD-150, (Released:2018-02-16)

被引用文献数

14

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Calcium is widely accepted as the main factor responsible for blossom-end rot (BER) appearance in tomato (Solanum lycopersicum L.) fruit. However, reactive oxygen species (ROS), which can damage plant tissues have also been proposed to initiate BER appearance in tomatoes and other fruit-bearing vegetables. Ascorbate, the major antioxidant in tomato fruit, is generally lower during green fruit development, which corresponds to the stage of BER appearance. Accordingly, one hypothesis is that tomato cultivars with a lower susceptibility to BER under salt stress have higher ascorbate contents and thus better control of ROS levels. In this study, to clarify the relationship between BER incidence and oxidative stress, two BER resistant cultivars, ‘Managua RZ’ and ‘House Momotaro’ and one BER-susceptible cultivar ‘Reiyoh’, were cultivated under salinity or standard nutrient solution (control) conditions. Calcium, potassium, magnesium, total hydro-soluble antioxidants, and ascorbate concentrations were measured in the distal pericarp 1 to 2 days prior to symptom appearance and during symptom appearance in healthy and affected fruits. When salt stress was applied, only BER-resistant cultivars showed a significant increase in ascorbate contents prior to BER appearance as compared with their levels under the control condition. In contrast, pre-BER Ca2+ concentrations did not associate with the BER susceptibility of each cultivar. Interestingly, ‘Reiyoh’ showed much higher K+/Ca2+ and (K++Mg2+)/Ca2+ ratios than the two other cultivars in healthy fruits due to a strong tendency towards lowered Ca2+ concentrations. A similar tendency was also observed in apple “bitter pit”. The ability to increase the fruit antioxidant capacity and maintain mineral balance under salt stress conditions may explain the resistance to BER development in highly resistant cultivars, probably by the avoidance of oxidative-induced cell necrosis and stabilization of the cell membranes, respectively.