Advancement of the cultivation and upscaling of photoautotrophic suspension cultures using Chenopodium rubrum as a case study

Segečová, Anna; Červený, Jan; Roitsch, Thomas

doi:10.1007/s11240-018-1441-6

Cited by 5 publications

(6 citation statements)

References 52 publications

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“…The PA cell suspension of Arabidopsis thaliana was obtained from [74]. It was maintained under axenic conditions in Erlenmeyer flasks under 21 °C as described in [75]. The culture was sub-cultured every 3 weeks by transferring into fresh and autoclaved Gamborg medium (G 0210, Duchefa) with added 2,4- d (1 mg L −1 ), pH = 5.7.…”

Section: Methodsmentioning

confidence: 99%

“…Routinely, cell suspension growth can be presented as change in packed cell volume, DW or turbidity [75]; however, these techniques require relatively large sample volumes. Therefore, we present a measure of viability inhibition (Table 1c).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, we present a measure of viability inhibition (Table 1c). The viability was measured microscopically after staining with 0.4% water solution of Trypan blue (Carl Roth, DE) after [75]. The per cent viability inhibition is calculated as VI = (1−(x/c))*100, where VI is the viability inhibition, x is an average viability of the treated group, and c stands for average viability of the control group.…”

Section: Methodsmentioning

confidence: 99%

“…The Fv/Fm was derived from a built-in quenching analysis under orange-red measuring light (617 nm) and blue saturation pulse (450 nm, 1070 µmol m −2 s −1 ). In order to avoid complications with rapid sedimentation of the suspension cultures, the FluorCam was used in an inverted set-up as described in [75]. The measurements were made after 1.5, 6, 24 and 48 h. The samples treated with chromium were also measured at 72 h. Prior to measurements, the PA cell suspension in the microplates was pre-darkened for 10 min.…”

Section: Methodsmentioning

confidence: 99%

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Noninvasive determination of toxic stress biomarkers by high-throughput screening of photoautotrophic cell suspension cultures with multicolor fluorescence imaging

et al. 2019

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Background With increasing pollution, herbicide application and interest in plant phenotyping, sensors capturing early responses to toxic stress are demanded for screening susceptible or resistant plant varieties. Standard toxicity tests on plants are laborious, demanding in terms of space and material, and the measurement of growth-inhibition based endpoints takes relatively long time. The aim of this work was to explore the potential of photoautotrophic cell suspension cultures for high-throughput early toxicity screening based on imaging techniques. The investigation of the universal potential of fluorescence imaging methods involved testing of three toxicants with different modes of action (DCMU, glyphosate and chromium). Results The increased pace of testing was achieved by using non-destructive imaging methods—multicolor fluorescence (MCF) and chlorophyll fluorescence (ChlF). These methods detected the negative effects of the toxicants earlier than it was reflected in plant growth inhibition (decrease in leaf area and final dry weight). Moreover, more subtle and transient effects not resulting in growth inhibition could be detected by fluorescence. The pace and sensitivity of stress detection was further enhanced by using photoautotrophic cell suspension cultures. These reacted sooner, more pronouncedly and to lower concentrations of the tested toxicants than the plants. Toxicant-specific stress signatures were observed as a combination of MCF and ChlF parameters and timing of the response. Principal component analysis was found to be useful for reduction of the collected multidimensional data sets to a few informative parameters allowing comparison of the toxicant signatures. Conclusions Photoautotrophic cell suspension cultures have proved to be useful for rapid high-throughput screening of toxic stress and display a potential for employment as an alternative to tests on whole plants. The MCF and ChlF methods are capable of distinguishing early stress signatures of at least three different modes of action.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Noninvasive determination of toxic stress biomarkers by high-throughput screening of photoautotrophic cell suspension cultures with multicolor fluorescence imaging

et al. 2019

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“…Similarly, T87 cells are derived from Col-0 plants and are maintained in liquid MS medium, where they also display rapid growth. Furthermore, both PSB and T87 cells ( Segečová et al., 2018 ; Li et al., 2020 ) have been shown to generate fully functional chloroplasts when grown in the light. The PSB-D and T87 cell lines are available as Arabidopsis Biological Resource Center stocks and are widely used in plant research, particularly for studying protein-protein interactions and recombinant protein expression ( Zhang et al., 2017 , 2020b ).…”

Section: Plant Cell Cultures As Bio-factoriesmentioning

confidence: 99%

Plant cell cultures as heterologous bio-factories for secondary metabolite production

Kerbler

Fernie

et al. 2021

Plant Communications

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Synthetic biology has been developing rapidly in the last decade and is attracting increasing attention from many plant biologists. The production of high-value plant-specific secondary metabolites is, however, limited mostly to microbes. This is potentially problematic because of incorrect post-translational modification of proteins and differences in protein micro-compartmentalization, substrate availability, chaperone availability, product toxicity, and cytochrome p450 reductase enzymes. Unlike other heterologous systems, plant cells may be a promising alternative for the production of high-value metabolites. Several commercial plant suspension cell cultures from different plant species have been used successfully to produce valuable metabolites in a safe, low cost, and environmentally friendly manner. However, few metabolites are currently being biosynthesized using plant platforms, with the exception of the natural pigment anthocyanin. Both Arabidopsis thaliana and Nicotiana tabacum cell cultures can be developed by multiple gene transformations and CRISPR-Cas9 genome editing. Given that the introduction of heterologous biosynthetic pathways into Arabidopsis and N. tabacum is not widely used, the biosynthesis of foreign metabolites is currently limited; however, therein lies great potential. Here, we discuss the exemplary use of plant cell cultures and prospects for using A. thaliana and N. tabacum cell cultures to produce valuable plant-specific metabolites.

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Gas exchange rates and sucrose concentrations affect plant growth and production of flavonoids in Vernonia condensata grown in vitro

Fortini

Batista

Mamedes-Rodrigues

et al. 2021

Plant Cell Tiss Organ Cult

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Advancement of the cultivation and upscaling of photoautotrophic suspension cultures using Chenopodium rubrum as a case study

Cited by 5 publications

References 52 publications

Noninvasive determination of toxic stress biomarkers by high-throughput screening of photoautotrophic cell suspension cultures with multicolor fluorescence imaging

Noninvasive determination of toxic stress biomarkers by high-throughput screening of photoautotrophic cell suspension cultures with multicolor fluorescence imaging

Plant cell cultures as heterologous bio-factories for secondary metabolite production

Gas exchange rates and sucrose concentrations affect plant growth and production of flavonoids in Vernonia condensata grown in vitro

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