Language：Japanese   Publishing type：Research paper (scientific journal)  

DOI： 10.11450/seitaikogaku.35.59

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Language：English   Publishing type：Research paper (international conference proceedings)   Publisher：International Society for Horticultural Science (ISHS)  

DOI： 10.17660/actahortic.2023.1364.1

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Language：English   Publishing type：Research paper (international conference proceedings)  

DOI： 10.1016/j.ifacol.2022.11.124

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.plaphy.2022.07.010

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Language：Japanese   Publishing type：Research paper (scientific journal)  

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Publishing type：Research paper (scientific journal)   Publisher：International Society for Horticultural Science (ISHS)  

DOI： 10.17660/actahortic.2022.1337.38

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Japanese Society of Agricultural, Biological and Environmental Engineers and Scientists  

DOI： 10.2525/ecb.60.13

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media LLC  

DOI： 10.1007/s11240-021-02037-y

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Other Link： http://link.springer.com/article/10.1007/s11240-021-02037-y/fulltext.html

Language：English   Publishing type：Research paper (scientific journal)  

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Language：Japanese   Publishing type：Research paper (scientific journal)  

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Society of Agricultural Meteorology of Japan  

DOI： 10.2480/agrmet.d-21-00029

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Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

DOI： 10.1016/j.scienta.2020.109550

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Publishing type：Research paper (scientific journal)  

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC ECO-ENGINEERING, ENV SCI & TECH, GRAD SCH LIFE & ENV SCI  

As a highly sophisticated strategy for environmental control in greenhouses, the concept of speaking plant approach (SPA) has attracted a great deal of attention. Sensor-based plant diagnosis techniques to monitor plant physiological status are the first and most important step in SPA. Currently, imaging systems for plant growth monitoring are especially required in wide range of agricultural production not only in advanced greenhouses but also in traditional vinyl houses. In this study, we developed a compact- and lightweight-multiple imaging system that is consist of an infrared camera filtered with a long-pass filter (lambda > 620 nm), a color camera, and blue/red/infrared/white LEDs. The developed imaging system enable us to conduct chlorophyll fluorescence imaging, NDVI imaging, and color imaging at the same time. We applied the system for imaging of tomato plant grown under greenhouse condition and successfully obtained an obvious chlorophyll fluorescence induction curve derived from a series of chlorophyll fluorescence images and NDVI and color images that are useful to identify the matured tomato fruits.

DOI： 10.11450/seitaikogaku.32.33

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Language：Japanese   Publishing type：Research paper (scientific journal)   Publisher：SOC ECO-ENGINEERING, ENV SCI & TECH, GRAD SCH LIFE & ENV SCI  

The Speaking Plant Approach (SPA) is regarded as a desirable concept which defines that the environmental factors should be adjusted to the crop's physiological status. The first and most important step in the SPA concept is to obtain physiological information from a living plant. Especially for environmental control in highly sophisticated greenhouse, daily measurement of plant growth must be indispensable in SPA concept. In our previous study, a robotized chlorophyll fluorescence (CF) imaging system that evaluates daily changes in photosynthetic function of tomato canopy has been developed and came onto the market in 2015. In this study, we applied the robotized CF imaging system to measure the daily stem elongation of tomato plants grown in greenhouse. CF images of shoot apex of tomato plants were taken by using the robotized CF imaging system and detected the shoot apex manually by using commercially available image analysis software to obtain the average plant height. Then, the daily stem elongation was calculated by subtracting the average plant height of one day before from that of the day. The performance verification test proved that the spatial resolution of CF image was 1.6 mm pixel(-1) and the reproducibility of average height measurement was within 0.01 %. And, a long-term stem elongation monitoring for consecutive 70 days, which was done with the conventional weekly stem elongation monitoring by using a measuring tape, suggested that daily stem elongation is able to be monitored with the robotized CF imaging system.

DOI： 10.11450/seitaikogaku.32.15

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：KOREAN SOC HORTICULTURAL SCIENCE  

DOI： 10.1007/s13580-018-0047-6

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：SOC BIOSCIENCE BIOENGINEERING JAPAN  

Transient gene expression in whole plants by using viral vectors is promising as a rapid, mass production system for biopharmaceutical proteins. Recent studies have indicated that plant growth conditions such as air temperature markedly influence the accumulation levels of target proteins. Here, we investigated time course of the amount of recombinant hemagglutinin (HA), a vaccine antigen of influenza virus, in leaves of Nicotiana benthamiana plants grown at 20 degrees C or 25 degrees C post viral vector inoculation. The HA content per unit of leaf biomass increased and decreased from 4 to 6 days post inoculation at 20 degrees C and 25 degrees C, respectively, irrespective of the subcellular localization of HA. The overall HA contents were higher when HA was targeted to the endoplasmic reticulum (ER) rather than the apoplast. Necrosis of leaf tissues was specifically observed in plants inoculated with the ER-targeting vector and grown at 25 degrees C. With the ER targeting vector, the maximum HA contents at 20 degrees C and 25 degrees C were recorded at 6 and 4 days post inoculation, respectively, and were comparable to each other. HA contents thereafter decreased at both temperatures; the rate of reduction appeared faster at 25 degrees C than at 20 degrees C. From a practical point of view, our results indicate that the strategy of targeting HA to the ER, growing plants at a lower temperature of 20 degrees C, and harvesting leaves at around a week after vector inoculation should be implemented to obtain a high HA yield stably and efficiently. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.

DOI： 10.1016/j.jbiosc.2017.04.007

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY  

Agrobacterium-mediated transient expression systems enable plants to rapidly produce a wide range of recombinant proteins. To achieve economically feasible upstream production and downstream processing, it is beneficial to obtain high levels of two yield-related quantities of upstream production: recombinant protein content per fresh mass of harvested biomass (ggFM(-1)) and recombinant protein productivity per unit area-time (gm(-2)/month). Here, we report that the density of Nicotiana benthamiana plants during upstream production had significant impacts on the yield-related quantities of recombinant hemagglutinin (HA). The two quantities were smaller at a high plant density of 400plants m(-2) than at a low plant density of 100plants m(-2). The smaller quantities at the high plant density were attributed to: (i) a lower HA content in young leaves, which usually have high HA accumulation potentials; (ii) a lower biomass allocation to the young leaves; and (iii) a high area-time requirement for plants. Thus, plant density is a key factor for improving upstream production in Agrobacterium-mediated transient expression systems. Biotechnol. Bioeng. 2017;114: 1762-1770. (c) 2017 Wiley Periodicals, Inc.

DOI： 10.1002/bit.26303

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：WILEY-BLACKWELL  

The use of detached leaves instead of whole plants provides an alternative means for recombinant protein production based on Agrobacterium tumefaciens-mediated transient gene overexpression. However, the process for high-level protein production in detached leaves has not yet been established. In this study, we focused on leaf handling and maintenance conditions immediately after infiltration with Agrobacterium suspension (agroinfiltration) to improve recombinant protein expression in detached Nicotiana benthamiana leaves. We demonstrated that the residual water of bacterial suspension in detached leaves had significant impact on the yield of recombinant influenza hemagglutinin (HA). Immediately after agroinfiltration, detached leaves were stored in a dehumidified chamber to allow bacterial suspension water occupying intercellular space to be removed by transpiration. We varied the duration of this water removal treatment from 0.7 to 4.4h, which resulted in leaf fresh weights ranging from 0.94 to 1.28gg(-1) relative to weights measured just before agroinfiltration. We used these relative fresh weights (RFWs) as an indicator of the amount of residual water. The detached leaves were then incubated in humidified chambers for 6 days. We found that the presence of residual water significantly decreased HA yield, with a clear inverse correlation observed between HA yield and RFW. We next compared HA yields in detached leaves with those obtained from intact leaves by whole-plant expression performed at the same time. The maximum HA yield obtained from a detached leaf with a RFW of approximately 1.0, namely, 800g gFW(-1), was comparable to the mean HA yield of 846g gFW(-1) generated in intact leaves. Our results indicate the necessity of removing bacterial suspension water from agroinfiltrated detached leaves in transient overexpression systems and point to a critical factor enabling the detached-leaf system as a viable recombinant protein factory. (c) 2015 Wiley Periodicals, Inc.

DOI： 10.1002/bit.25854

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Frontiers Media S.A.  

Agrobacterium-mediated transient expression systems enable plants to produce a wide range of recombinant proteins on a rapid timescale. To achieve economically feasible upstream production and downstream processing, two yield parameters should be considered: (1) recombinant protein content per unit biomass and (2) recombinant protein productivity per unit area-time at the end of the upstream production. Because environmental factors in the upstream production have impacts on these parameters, environment control is important to maximize the recombinant protein yield. In this review, we summarize the effects of pre- and postinoculation environmental factors in the upstream production on the yield parameters and discuss the basic concept of environment control for plant-based transient expression systems. Preinoculation environmental factors associated with planting density, light quality, and nutrient supply affect plant characteristics, such as biomass and morphology, which in turn affect recombinant protein content and productivity. Accordingly, environment control for such plant characteristics has significant implications to achieve a high yield. On the other hand, postinoculation environmental factors, such as temperature, light intensity, and humidity, have been shown to affect recombinant protein content. Considering that recombinant protein production in Agrobacterium-mediated transient expression systems is a result of a series of complex biological events starting from T-DNA transfer from Agrobacterium tumefaciens to protein biosynthesis and accumulation in leaf tissue, we propose that dynamic environment control during the postinoculation process, i.e., changing environmental conditions at an appropriate timing for each event, may be a promising approach to obtain a high yield. Detailed descriptions of plant growth conditions and careful examination of environmental effects will significantly contribute to our knowledge to stably obtain high recombinant protein content and productivity, thus enhancing the utility of plant-based transient expression systems as recombinant protein factories.

DOI： 10.3389/fbioe.2016.00023

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：JAPANESE SOC PLANT CELL & MOLECULAR BIOLOGY  

Viral vector-mediated transient expression enables rapid biopharmaceutical protein production using whole plants. In recent years, plant growth conditions have received close attention as factors influencing the content of recombinant protein. In particular, because nitrogen is an important resource used in protein synthesis, nitrogen supplied to the root zone may influence accumulation of recombinant proteins. We tested whether fertilization of Nicotiana benthamiana with high-nitrate nutrient solution during plant growth prior to Agrobacterium-mediated delivery (agroinfiltration) of a tobamoviral vector resulted in high recombinant influenza hemagglutinin (HA) content per unit of leaf fresh weight and/or per plant. Nutrient solution containing 60 mM nitrate yielded 40% higher HA content per unit of leaf fresh weight, and comparable HA content per plant, relative to nutrient solution containing 12 or 36 mM nitrate. There were positive correlations among HA, total soluble protein (TSP), and soluble reduced-nitrogen content, suggesting that fertilization with the high-nitrate nutrient solution increased soluble reduced nitrogen and subsequently provided high TSP and HA. Although the increased HA content per unit of leaf fresh weight was offset by reduced leaf fresh weight on a per plant basis, nitrate-enriched fertilization represents an easily applicable technique to obtain a given amount of HA from smaller leaf biomass, thereby potentially reducing downstream processing costs for HA vaccine production using viral vector-mediated transient expression systems.

DOI： 10.5511/plantbiotechnology.14.0403a

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Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)  

We investigated effects of dim light irradiation (about 11 fimo\\ irT2 s ' photon flux density) of the root zone on the growth and development of leaf lettuce (Lactuca sativa 'Okayama saradana') plants under nutrient film technique (NFT) hydroponics with monochromatic light emitting diodes (LEDs) of five types with respective peak wavelengths (and colors) of 405 nm (violet), 465 nm (blue), 525 nm (green), 660 nm (red), and 735 nm (far-red). Shoot fresh weight and specific root length in the treatments of dim light irradiation of the root zone with violet, blue, and far-red LEDs were significantly lower than in the control (no irradiation of the root zone). Roots in these treatments developed thickly, probably because of suppressed lateral root initiation because sparsely distributed roots were observed in these treatments. Dim light irradiation of the root zone of leaf lettuce plants did not affect mass production, but it did affect root morphology.

DOI： 10.2525/ecb.50.101

Scopus

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Event date： 2024.5

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Event date： 2019.6

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Venue：China Beijing  

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Venue：日本 鹿児島  

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Venue：日本 東京  

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Venue：日本 奈良  

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Venue：日本 松山  

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Faculty of Agriculture / Specialized Subjects

Faculty of Agriculture

農学部

Faculty of Agriculture

Graduate School of Agriculture

Faculty of Agriculture

Faculty of Agriculture / Specialized Subjects

農学研究科（修士課程）

Faculty of Agriculture

Faculty of Agriculture

Graduate School of Agriculture

Type：Seminar, workshop

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Type：Academic society, research group, etc. 

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