MY Research

(2022.01.06 updated)

Environment Control for Plant-made pharmaceutical Protein Production

We focus on the production of recombinant proteins using plants as one of the effective applications of plant factories with artificial lighting (PFALs). A typical example is pharmaceutical proteins including vaccine antigens and monoclonal antibodies. Conventionally, these pharmaceutical proteins have been produced by culturing microorganisms or mammalian cells, or by culturing viruses using embryonated chicken eggs. On the other hand, the use of plants is thought to offer advantages such as lower production costs, higher scalability of production to meet demand, and reduced risk of pathogen contamination during the production process.

We have conducted basic research to establish the environmental control strategy for PFALs or greenhouses suitable for the production of recombinant proteins using plants. In particular, we have been targeting the transient gene expression method, in which genes encoding recombinant proteins are introduced into mature plants (Nicotiana benthamiana) to express transiently. We would like to contribute to the utilization and dissemination of this new plant biotechnology from the standpoint of bioenvironmental engineering.

Related publications:

Fujiuchi, N., N. Matoba, K. Fujiwara, R. Matsuda. (2021) Effects of lighting conditions on Agrobacterium-mediated transient expression of recombinant hemagglutinin in detached Nicotiana benthamiana leaves inoculated with a deconstructed viral vector. Plant Cell, Tissue and Organ Culture 145(3): 679–688.

Matsuda, R. (2021) A 24-h photoperiod before gene transfer promotes biomass production but not the yield of influenza hemagglutinin transiently expressed in Nicotiana benthamiana. Journal of Agricultural Meteorology 77(4): 278–283.

Matsuda, R., A. Ueno, K. Fujiwara. (2019) Effects of environmental conditions before gene transfer on the amount of influenza hemagglutinin transiently expressed in Nicotiana benthamiana leaves. Journal of Agricultural Meteorology 75(3): 129–136.

Matsuda, R., A. Ueno, H. Nakaigawa, K. Fujiwara. (2018) Gas exchange rates decrease and leaf temperature increases in Nicotiana benthamiana leaves transiently overexpressing hemagglutinin in an Agrobacterium-assisted viral vector system. Frontiers in Plant Science 9: 1315.

Fujiuchi, N., R. Matsuda, N. Matoba, K. Fujiwara. (2017) Effects of plant density on recombinant hemagglutinin yields in an Agrobacterium-mediated transient gene expression system using Nicotiana benthamiana plants. Biotechnology and Bioengineering 114(8): 1762–1770.

Matsuda, R., T. Abe, N. Fujiuchi, N. Matoba, K. Fujiwara. (2017) Effect of temperature post viral vector inoculation on the amount of hemagglutinin transiently expressed in Nicotiana benthamiana leaves. Journal of Bioscience and Bioengineering 124(3): 346–350.

Matsuda, R., T. Abe, K. Fujiwara. (2017) Viral vector-based transient gene expression in Nicotiana benthamiana: effects of light source on leaf temperature and hemagglutinin content. Plant Cell Reports 36(10): 1667–1669.

Fujiuchi, N., N. Matoba, R. Matsuda. (2016) Environment control to improve recombinant protein yields in plants based on Agrobacterium-mediated transient gene expression. Frontiers in Bioengineering and Biotechnology 4: 23.

Fujiuchi, N., R. Matsuda, N. Matoba, K. Fujiwara. (2016) Removal of bacterial suspension water occupying the intercellular space of detached leaves after agroinfiltration improves the yield of recombinant hemagglutinin in a Nicotiana benthamiana transient gene expression system. Biotechnology and Bioengineering 113(4): 901–906.

Fujiuchi, N., R. Matsuda, N. Matoba, K. Fujiwara. (2014) Effect of nitrate concentration in nutrient solution on hemagglutinin content of Nicotiana benthamiana leaves in a viral vector-mediated transient gene expression system. Plant Biotechnology 31(3): 207–211.

Matsuda, R., A. Tahara, N. Matoba, K. Fujiwara. (2012) Virus-vector mediated rapid protein production in Nicotiana benthamiana: effects of temperature and photosynthetic photon flux density on hemagglutinin accumulation. Environment Control in Biology 50(4): 375–381.

Matsuda, R., C. Kubota, M.L. Alvarez, G.A. Cardineau. (2012) Effect of high electrical conductivity of hydroponic nutrient solution on vaccine protein content in transgenic tomato. HortTechnology 22(3): 362–367.

Matsuda, R., C. Kubota. (2010) Variation of total soluble-protein content in fruit among six greenhouse tomato cultivars. HortScience 45(11): 1645–1648.

Matsuda, R., C. Kubota, M.L. Alvarez, G.A. Cardineau. (2010) Determining the optimal timing of fruit harvest in transgenic tomato expressing F1-V, a candidate subunit vaccine against plague. HortScience 45(3): 347–351.

Matsuda, R., C. Kubota, M.L. Alvarez, G.A. Cardineau. (2009) Biopharmaceutical protein production under controlled environments: growth, development, and vaccine productivity of transgenic tomato plants grown hydroponically in a greenhouse. HortScience 44(6): 1594–1599.

Response of Photosynthesis and growth of crop plants to Light Environment and Application to Light Environment Control in Horticulture

This project aims to analyze the effects of the light environment on photosynthesis and plant growth from an ecophysiological point of view and to apply the findings to the appropriate control of the light environment in greenhouses and plant factories with artificial lighting. So far, we have been studying the effects of spectral distribution (light quality), supplemental lighting to lower leaves, continuous (24-h) lighting, etc. Currently, we are interested in the effects of temporally fluctuating light environments. For these studies, we have utilized LEDs since we started to work nearly 20 years ago, which have unique features to easily control spectral distribution and the time course of power output and to irradiate locally. By using the knowledge and techniques of both biology and engineering, we aim to acquire interesting findings and develop new technologies in agriculture.

Related publications:

Chen, Y., R. Matsuda, K. Fujiwara. (2022) Rapid and semi-automated leaf net photosynthetic rate determination for numerous phosphor-converted white-LED lights of different spectral distributions. Journal of Agricultural Meteorology 78(1): 8–18.

Yu, L., K. Fujiwara, R. Matsuda. (2022) Recent levels of daily photosynthetic photon flux density have a larger influence on light acclimation responses of cucumber leaves than early levels. Horticulture, Environment, and Biotechnology in press.

Yu, L., K. Fujiwara, R. Matsuda. (2022) Estimating light acclimation parameters of cucumber leaves using time-weighted averages of daily photosynthetic photon flux density. Frontiers in Plant Science in press.

Jishi, T., R. Matsuda, K. Fujiwara. (2021) Blue light monochromatic irradiation for 12 h in lighting pattern with combinations of blue and red light elongates young cos lettuce leaves and promotes growth under high daily light integral. HortScience 56(8) 940–945.

Jishi, T., R. Matsuda, K. Fujiwara. (2021) Manipulation of intraday durations of blue- and red-light irradiation to improve cos lettuce growth. Frontiers in Plant Science 12: 778205.

Matsuda, R., H. Ito, K. Fujiwara. (2021) Effects of artificially reproduced fluctuations in sunlight spectral distribution on the net photosynthetic rate of cucumber leaves. Frontiers in Plant Science 12: 675810.

Jishi, T., R. Matsuda, K. Fujiwara. (2018) Effects of photosynthetic photon flux density, frequency, duty ratio and their interactions on net photosynthetic rate of cos lettuce leaves under pulsed light: explanation based on photosynthetic-intermediate pool dynamics. Photosynthesis Research 136(3): 371–378.

Murakami, K., R. Matsuda, K. Fujiwara. (2018) Quantification of excitation energy distribution between photosystems based on a mechanistic model of photosynthetic electron transport. Plant, Cell and Environment 41(1): 148–159.

Murakami, K., R. Matsuda, K. Fujiwara. (2018) A mathematical model of photosynthetic electron transport in response to light spectrum based on excitation energy distributed to photosystems. Plant and Cell Physiology 59(8): 1643–1651.

Murakami, K., R. Matsuda, K. Fujiwara. (2017) A basis for selecting light spectral distribution for evaluating leaf photosynthetic rates of plants grown under different light spectral distributions. Environmental Control in Biology 55(1): 1–6.

Jishi, T., K. Kimura, R. Matsuda, K. Fujiwara. (2016) Effects of temporally shifted irradiation of blue and red LED light on cos lettuce growth and morphology. Scientia Horticulturae 198: 227–232.

Matsuda, R., T. Yamano, K. Murakami, K. Fujiwara. (2016) Effects of spectral distribution and photosynthetic photon flux density for overnight LED light irradiation on tomato seedling growth and leaf injury. Scientia Horticulturae 198: 363–369.

Murakami, K., R. Matsuda, K. Fujiwara. (2016) Interaction between the spectral photon flux density distributions of light during growth and for measurements in net photosynthetic rates of cucumber leaves. Physiologia Plantarum 158(2): 213–224.

Jishi, T., R. Matsuda, K. Fujiwara. (2015) A kinetic model for estimating net photosynthetic rates of cos lettuce leaves under pulsed light. Photosynthesis Research 124(1): 107–116.

Matsuda, R., N. Ozawa, K. Fujiwara. (2014) Leaf photosynthesis, plant growth, and carbohydrate accumulation of tomato under different photoperiods and diurnal temperature differences. Scientia Horticulturae 170: 150–158.

Murakami, K., R. Matsuda, K. Fujiwara. (2014) Light-induced systemic regulation of photosynthesis in primary and trifoliate leaves of Phaseolus vulgaris: effects of photosynthetic photon flux density (PPFD) versus spectrum. Plant Biology 16(1): 16–21.

Murakami, K., R. Matsuda, K. Fujiwara. (2013) Effects of supplemental lighting to a lower leaf using light-emitting diodes with different spectra on the leaf photosynthetic rate in sweet pepper. Journal of Agricultural Meteorology 69(2): 55–63.

Matsuda, R., K. Ohashi-Kaneko, K. Fujiwara, K. Kurata. (2008) Effects of blue light deficiency on acclimation of light energy partitioning in PSII and CO₂ assimilation capacity to high irradiance in spinach leaves. Plant and Cell Physiology 49(4): 664–670.

Matsuda, R., K. Ohashi-Kaneko, K. Fujiwara, K. Kurata. (2007) Analysis of the relationship between blue-light photon flux density and the photosynthetic properties of spinach (Spinacia oleracea L.) leaves with regard to the acclimation of photosynthesis to growth irradiance. Soil Science and Plant Nutrition 53(4): 459–465.

Ohashi-Kaneko, K., R. Matsuda, E. Goto, K. Fujiwara, K. Kurata. (2006) Growth of rice plants under red light with or without supplemental blue light. Soil Science and Plant Nutrition 52(4): 444–452.

Matsuda, R., K. Ohashi-Kaneko, K. Fujiwara, E. Goto, K. Kurata. (2004) Photosynthetic characteristics of rice leaves grown under red light with or without supplemental blue light. Plant and Cell Physiology 45(12): 1870–1874.