Effects of lighting conditions on Agrobacterium-mediated transient expression of recombinant hemagglutinin in detached Nicotiana benthamiana leaves inoculated with a deconstructed viral vector

• Published in: Plant cell, tissue and organ culture Vol. 145; no. 3; pp. 679 - 688
• Main Authors: Fujiuchi, Naomichi, Matoba, Nobuyuki, Fujiwara, Kazuhiro, Matsuda, Ryo
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01-06-2021; Springer Nature B.V
• Subjects: Agrobacterium; Biomedical and Life Sciences; Detaching; Energy conservation; Environmental control; Flux density; Gene expression; Hemagglutinins; Influenza; Leaves; Life Sciences; Light sources; Lighting; Low cost; Nicotiana benthamiana; Original Article; Photosynthesis; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant Sciences; Proteins; Vectors (Biology); Molecular farming; Plant-made pharmaceuticals; Environment control; Viral vector; Tobacco plants
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-021-02037-y

Environmental control of plant incubation conditions before and after agroinfiltration is crucial during recombinant protein production using an
Agrobacterium
-mediated transient gene expression system to ensure high and consistent productivity. In an effort to design a low-cost and energy-saving upstream production process, we assessed the lighting effects during the post-agroinfiltration process on the leaf content of recombinant influenza hemagglutinin (HA) expressed by the magnICON® deconstructed viral vector system. Detached
Nicotiana benthamiana
leaves were incubated for 6 days post-agroinfiltration under five different lighting conditions: (1) 6-day dark (D–D), (2) 3-day dark followed by 3-day lighting at a photosynthetic photon flux density (PPFD) of 100 µmol m
−2
 s
−1
(D–L100) and (3) its reversed lighting condition (L100–D), and (4) 6-day lighting at a PPFD of 100 µmol m
−2
 s
−1
(L100–L100) and (5) 500 µmol m
−2
 s
−1
(L500–L500). The HA content at 6 days post-agroinfiltration was 25 µg gFM
−1
(D–D), 408 µg gFM
−1
(D–L100), 566 µg gFM
−1
(L100–L100), 693 µg gFM
−1
(L500–L500) and 756 µg gFM
−1
(L100–D), respectively. The HA content in L100–D was not less than that in the other lighting conditions, indicating that lighting during the latter half of the post-agroinfiltration process is not crucial for recombinant protein production. Based on these findings, we propose to implement a multiphase environment control in the upstream production process of transient expression system, wherein reduced number of light sources may provide a low-cost and energy-saving design for the post-agroinfiltration phase.
Key message
Post-agroinfiltration 3-day lighting and subsequent 3-day dark enabled detached
Nicotiana benthamiana
leaves to produce and accumulate a high amount of recombinant influenza hemagglutinin using the magnICON® transient expression system.