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Monitoring of Plant Cells and Tissues in Bioprocesses

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Bioprocessing of Plant In Vitro Systems

Part of the book series: Reference Series in Phytochemistry ((RSP))

Abstract

Plant cell and tissue cultures represent a suitable alternative as production systems for valuable plant secondary metabolites. Unlike traditional extraction from agricultural grown plants, the active ingredient production in biotechnological processes with in vitro cultures takes place in closed bioreactors under controlled conditions. This allows a year-round production with constant quality and quantity. However, the development of biotechnological processes with plant in vitro cultures is often time-consuming and requires parallelized screening systems. Furthermore, the design, optimization, and control of economic processes presuppose knowledge about the physiological state of the biological system and the kinetic parameters of biomass and product formation. To gain access to these data, suitable process-monitoring methods are required which provide information about the physiology of the process, both on a macroscopic and on the single cell level. However, due to the morphology of plant cell and tissue cultures, many methods for bioprocess monitoring that are used for mammalian and microbial cultures are not applicable. This chapter covers methods that are appropriate for monitoring of biotechnological processes with plant cell and tissue cultures: The conductivity of the growth medium is a powerful parameter to estimate the growth of complex plant cell aggregates and tissue structures. The next section describes the application of the RAMOS – a small scale cultivation system – for heterotrophic and phototrophic plant cell and tissue cultures. Flow cytometry is a tool to obtain segregated data of bioprocesses. Further, we describe a novel approach of cell immobilization for physiological studies and the design of bioprocesses, the 3D Green Bioprinting.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

BrdU:

Bromodeoxyuridine

CAD:

Computer-aided design

CAM:

Computer-aided manufacturing

cbulk:

Concentration of substrates/products in culture medium [g l−1]

CLSM:

Confocal laser scanning microscopy

cpore:

Concentration of substrates/products in a hydrogel pore [g l−1]

cs:

Cell suspension culture

CT:

Carbon dioxide transfer [mmol l−1]

CTR:

Carbon dioxide transfer rate [mmol l−1 h−1]

CTRev:

Evaporation-corrected carbon dioxide transfer rate [mmol l−1 h−1]

ctrmax:

Maximum biomass-specific carbon dioxide transfer rate [mmol g−1 h−1]

CTRmax:

Maximum carbon dioxide transfer rate [mmol l−1 h−1]

C-value:

DNA content of the holoploid genome with chromosome number n

Cx-value:

DNA content of the monoploid genome with chromosome number x

DW:

Concentration of biomass dry weight [g l−1]

DWmax:

Maximum concentration of biomass dry weight [g l−1]

EdU:

5-Ethynyl-2′-deoxyuridine

Fev:

Rate of evaporation [ml h−1]

FUCCI:

Fluorescent ubiquitination-based cell-cycle indicator

G0/G1 phase:

Cell cycle phase

G2/M phase:

Cell cycle phase

HPLC:

High performance liquid chromatography

hr:

Hairy root culture(s)

LED:

Light emitting diode(s)

LS:

Linsmaier and Skoog medium

MS:

Murashige and Skoog medium

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

n:

Nuclear phase status:

n – means the chromosome number of the meiotically reduced genome irrespective of the ploidy

2n – means the chromosome number of the non-reduced genome

N:

Number of data points

OLED:

Organic light emitting diode(s)

OT:

Oxygen transfer [mmol l−1]

OTR:

Oxygen transfer rate [mmol l−1 h−1]

OTRev:

Evaporation-corrected oxygen transfer rate [mmol l−1 h−1]

otrmax:

Maximum biomass-specific oxygen transfer rate [mmol g−1 h−1]

OTRmax:

Maximum oxygen transfer rate [mmol l−1 h−1]

OU:

Oxygen uptake [mmol l−1]

OUR:

Oxygen uptake rate [mmol l−1 h−1]

p:

Overall pressure [bar]

PAR:

Photosynthetic active radiation

PBR:

Photobioreactor(s)

\( {\mathrm{p}}_{{\mathrm{CO}}_2} \) :

Carbon dioxide partial pressure [bar]

PFD:

Photon flux density [μmol m−2 s−1]

\( {\mathrm{p}}_{{\mathrm{O}}_2} \) :

Oxygen partial pressure [bar]

R:

Universal gas constant (0.08314 bar l mol−1 K−1)

RAMOS®:

Respiration Activity MOnitoring System®

RQ:

Respiration quotient

S:

Concentration of substrate [g l−1]

S phase:

Cell cycle phase

SEM:

Scanning electron microscope

t:

Time [d]

VL:

Initial liquid filling volume [ml]

Vt:

Total flask volume [ml]

x:

Number of chromosomes of the monoploid genome

α:

Proportionality constant

κ:

Conductivity [mS cm−1]

κm:

Measured conductivity [mS cm−1]

μ:

Specific growth rate

μmax:

Maximum specific growth rate

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Acknowledgment

The authors thank Joachim Püschel and Dr. Beatrice Weber from the Institute of Botany, TU Dresden, for helpful discussions concerning the scheme in Fig. 10.

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Authors and Affiliations

  1. Institute of Natural Materials Technology, Technische Universität Dresden, Bergstr. 120, 01062, Dresden, Germany

    Juliane Steingroewer, Christiane Haas, Katja Winkler, Carolin Schott, Jost Weber, Julia Seidel, Felix Krujatz, Sibylle Kümmritz & Maria Lisa Socher

  2. Bioprocess Engineering, Institute of Food Technology and Bioprocess Engineering, TU Dresden, Dresden, Germany

    Thomas Bley

  3. Evolva Copenhagen, Lersø Parkallé 42-44, 2100, Copenhagen, Denmark

    Jost Weber

  4. Centre for Translational Bone, Joint and Soft Tissue Research, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, 01062, Dresden, Germany

    Julia Seidel, Anja Lode & Michael Gelinsky

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  1. Juliane Steingroewer
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Correspondence to Juliane Steingroewer .

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  1. University of Food Technologies, Plovdiv, Bulgaria

    Atanas Pavlov

  2. Bioprocess Engineering, Institute of Food Technology and Bioprocess Engineering, TU Dresden, Dresden, Germany

    Thomas Bley

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Steingroewer, J. et al. (2018). Monitoring of Plant Cells and Tissues in Bioprocesses. In: Pavlov, A., Bley, T. (eds) Bioprocessing of Plant In Vitro Systems. Reference Series in Phytochemistry. Springer, Cham. https://doi.org/10.1007/978-3-319-54600-1_7

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