Effects of CO2 enrichment, LED inter-lighting, and high plant density on growth of Nicotiana benthamiana used as a host to express influenza virus hemagglutinin H1

Liu, Shang; Gaudreau, L.; Martel, Michèle; Michaud, Dominique; Pépin, Steeve; Gosselin, André

doi:10.1007/s13580-018-0085-0

Cited by 14 publications

(15 citation statements)

References 39 publications

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“…3c, d, Table 4 . Similar results of negligible effects of CO 2 enrichment were reported by Shang et al 2018 . Thus, no clear effects dependent upon CO 2 enrichment before gene transfer were found under the conditions tested in the present experiment.…”

Section: Effects Of Co 2 Concentrationsupporting

confidence: 90%

“…These results for leaf HA content per unit FM are consistent with results from our recent study in which leaf HA content per unit FM was significantly lowered when plant density was high 400 plants m 2 , Fujiuchi et al, 2017 , as dense planting decreases the amount of light received by a plant. In contrast, Shang et al 2018 reported that supplemental lighting with LEDs and dense planting in a greenhouse before gene transfer had little or no effect on leaf HA content per unit of biomass. Effects of light intensity may be different between indoor and greenhouse conditions and / or between transgene expression systems used.…”

Section: Effects Of Ppfdmentioning

confidence: 93%

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Effects of environmental conditions before gene transfer on the amount of influenza hemagglutinin transiently expressed in <I>Nicotiana benthamiana</I> leaves

Matsuda

Ueno

Fujiwara

2019

J. Agric. Meteorol.

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We investigated the effects of photosynthetic photon flux density PPFD , air temperature, and CO 2 concentration before gene transfer on the accumulation level of hemagglutinin HA , an influenza vaccine antigen, in Nicotiana benthamiana leaves in a transient gene expression system. Plants were treated for two weeks before gene transfer with different levels of PPFD, air temperature or CO 2 concentration in separate experiments. Leaf fresh mass FM , leaf HA content per unit FM, and leaf HA content per plant at 6 d post infiltration 6 DPI for gene transfer all tended to increase with increasing average PPFD during the treatment period. Leaf FM and leaf HA content per plant at 6 DPI were significantly greater at an average day air temperature of 25 C-30 C during the treatment period than at 20 C-25 C. No clear effects of CO 2 concentration were found on either growth or leaf HA content under the conditions tested in the present study. There was a positive correlation between leaf HA content per unit FM at 6 DPI and leaf FM at 0 or 6 DPI in plants grown under different PPFDs and air temperatures. Preparing plants with increased leaf FM by applying high PPFD and / or high air temperature before gene transfer should increase not only total HA yield per plant but also HA content per unit leaf biomass at harvest.

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Section: Effects Of Co 2 Concentrationsupporting

confidence: 90%

Section: Effects Of Ppfdmentioning

confidence: 93%

Effects of environmental conditions before gene transfer on the amount of influenza hemagglutinin transiently expressed in <I>Nicotiana benthamiana</I> leaves

Matsuda

Ueno

Fujiwara

2019

J. Agric. Meteorol.

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“…Cultural conditions have a significant impact on recombinant protein yields in N. benthamiana (Fujiuchi et al., 2014, 2017; Matsuda et al., 2017, 2018; Shang et al., 2018). Our goal in this study was to document eventual links between the effects of some current cultural practices on plant development and the production yield of influenza virus hemagglutinin H1, a promising vaccine antigen, in leaves of N. benthamiana used as a host for protein expression.…”

Section: Resultsmentioning

confidence: 99%

“…In practice, recombinant protein yield in plant (e.g., N. benthamiana ) transient expression settings depends on the net amount of protein per gram of leaf tissue; the leaf biomass per plant prior to agroinfiltration; and the number of plants per growing area before plant tissue downstream processing (Fujiuchi et al., 2016; Shang et al., 2018). Several studies have reported the development of molecular tools or strategies to improve recombinant protein yield and quality in N. benthamiana leaves, notably to maximize the structural resemblance between plant-made proteins and their original counterparts or to protect those proteins that show limited stability in plant cell environments (Faye et al., 2005; Gomord et al., 2010).…”

Section: Introductionmentioning

confidence: 99%

“…Other studies have described the expression of a viral proton channel to stabilize labile proteins in the cell secretory pathway (Jutras et al., 2015, 2018), the expression of a human convertase to promote the post-translational proteolytic processing of clinically useful proteins in vivo (Wilbers et al., 2016; Mamedov et al., 2019), or the exogenous induction of the jasmonic acid defense pathway to reduce endogenous protein content in leaf tissue prior to recombinant protein purification (Robert et al., 2015). In parallel, studies have documented the effects of cultural practices on N. benthamiana growth and leaf biomass production before agroinfiltration (Fujiuchi et al., 2014; Shang et al., 2018), the influence of environmental parameters in growth chambers following agroinfiltration (Matsuda et al., 2017, 2018), or the impact of plant density on overall protein yield in specific culture settings (Fujiuchi et al., 2017; Shang et al., 2018).…”

Section: Introductionmentioning

confidence: 99%

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Production of Biopharmaceuticals in Nicotiana benthamiana—Axillary Stem Growth as a Key Determinant of Total Protein Yield

et al. 2019

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Data are scarce about the influence of basic cultural conditions on growth patterns and overall performance of plants used as heterologous production hosts for protein pharmaceuticals. Higher plants are complex organisms with young, mature, and senescing organs that show distinct metabolic backgrounds and differ in their ability to sustain foreign protein expression and accumulation. Here, we used the transient protein expression host Nicotiana benthamiana as a model to map the accumulation profile of influenza virus hemagglutinin H1, a clinically promising vaccine antigen, at the whole plant scale. Greenhouse-grown plants submitted to different light regimes, submitted to apical bud pruning, or treated with the axillary growth-promoting cytokinin 6-benzylaminopurine were vacuum-infiltrated with agrobacteria harboring a DNA sequence for H1 and allowed to express the viral antigen for 7 days in growth chamber under similar environmental conditions. Our data highlight the importance of young leaves on H1 yield per plant, unlike older leaves which account for a significant part of the plant biomass but contribute little to total antigen titer. Our data also highlight the key contribution of axillary stem leaves, which contribute more than 50% of total yield under certain conditions despite representing only one-third of the total biomass. These findings underline the relevance of both considering main stem leaves and axillary stem leaves while modeling heterologous protein production in N. benthamiana. They also demonstrate the potential of exogenously applied growth-promoting hormones to modulate host plant architecture for improvement of protein yields.

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Synergetic light and cytokinin treatments mitigate the recombinant protein yield depression induced by high‐density cultivation of hydroponically‐grown Nicotiana benthamiana

Bilotta,

Éthier,

Laliberté

et al. 2024

Biotech & Bioengineering

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Plant molecular farming is currently operating a transition from soil‐based cultures toward hydroponic systems. In this study, we designed a whole‐plant NFT (nutrient film technique) platform for the transient expression of influenza virus‐like particles harboring hemagglutinin H1 proteins in Nicotiana benthamiana. In particular, we examined the effects of plant density during the post‐infiltration expression phase on plant growth and H1 yield in relation to the daily light integral (DLI) received by the crop and the exogenous application of 6‐BAP cytokinin (CK). We expected from previous work that high DLI and CK treatments would stimulate the development of highly productive leaves on axillary (secondary) stems and thereby improve the H1 yield at the whole‐plant scale. Increasing plant density from 35.7 to 61 plants m–2 during the post‐infiltration phase significantly decreased the proportion of axillary leaf biomass by 30% and H1 yield per plant by 39%, resulting in no additional yield gain on a whole‐crop area basis. Adding CK to the recirculated nutrient solution decreased the harvested leaf biomass by 31% and did not enhance the relative proportion of S leaves of the plants as previously reported with foliar CK application. There was a 36% increase in H1 yield when doubling the DLI from 14 to 28 mol m–2 s–1, and up to 71% yield gain when combining such an increase in DLI with the hydroponic CK treatment. Contrary to our expectations, leaves located on the main stem, particularly those from the upper half of the plant (i.e., eighth leaf and above), contributed about 80% of total H1 yield. Our study highlights the significantly different phenotype (~30% less secondary leaf biomass) and divergent responses to light and CK treatments of NFT‐grown N. benthamiana plants compared to previous studies conducted on potted plants.

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Effects of CO2 enrichment, LED inter-lighting, and high plant density on growth of Nicotiana benthamiana used as a host to express influenza virus hemagglutinin H1

Cited by 14 publications

References 39 publications

Effects of environmental conditions before gene transfer on the amount of influenza hemagglutinin transiently expressed in <I>Nicotiana benthamiana</I> leaves

Effects of environmental conditions before gene transfer on the amount of influenza hemagglutinin transiently expressed in <I>Nicotiana benthamiana</I> leaves

Production of Biopharmaceuticals in Nicotiana benthamiana—Axillary Stem Growth as a Key Determinant of Total Protein Yield

Synergetic light and cytokinin treatments mitigate the recombinant protein yield depression induced by high‐density cultivation of hydroponically‐grown Nicotiana benthamiana

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