Plant cell and hairy root cultures in bioreactor-based production processes

Dissertation

    Research output: ThesisDissertationCollection of Articles

    Abstract

    In this study, laboratory scale biotechnical production processes were developed for different types of plant cell cultures: embryogenic cell culture, undifferentiated cell suspension culture and hairy root culture. The production of somatic embryos, secondary metabolites and biomass were optimized. The growth as well as the production characteristics and the nutrient uptake of the different plant cell systems were determined. The suitabilities of different bioreactor configurations for the various culture types were tested in laboratory scale. In the optimization of growth and production, statistical experimental designs were used in addition to the conventional method of parallel testing of different media were used. The embryo production medium for an embryogenic cell line of birch (Betula pendula Roth) was optimized using a 22-factorial design. Factorial design was also tested for the optimization of alkaloid production medium for a cell suspension of Catharanthus roseus (L.) G. Don, but the conventional method of parallel testing of different media proved to be more successful. Factorial experimental design was used to determine the effects of low phosphate and sugar concentrations on strawberry (Fragaria x ananassa Duch.) hairy roots. In this way a slowly growing root system was obtained for infection studies with the native Finnish arbuscular mycorrhizal fungus, Glomus fistulosum V128. The embryogenic cell line of birch was cultivated in four different types of bioreactors: a sparged blade-stirred bioreactor, a diffusion-aerated bioreactor with a marine impeller (ChemCell, Chemap), an air-lift and a bubble column bioreactor. Embryo development was monitored during the cultivations by computer vision. The highest embryo production was achieved in the bubble column bioreactor. However, this was only 20% of the production in control shake flasks. In the mechanically stirred bioreactors production was even lower, indicating shear damage of the embryos. The suspension culture of Catharanthus roseus L. was cultivated in three different types of bioreactors: a standard aerated and stirred bioreactor (control bioreactor), a recirculation bioreactor in which part of the air is recirculated with a peristaltic pump, and a bubble column bioreactor. The recirculation bioreactor was used to mimic the gaseous regime of a shake flask. When cultured in a turbine stirred tank reactor aerated through a sparger and in the bubble column the cells did not grow well and the production of ajmalicine was low. When cultivated in the recirculation bioreactor the biomass concentration and production of ajmalicine were similar to the level observed in shake flasks. Hairy roots of C. roseus were cultivated in three different types of bioreactors: an air-sparged bioreactor with no stirring, an air-sparged bioreactor in which the medium is slowly circulated through the vessel with a peristaltic pump and a standard air-sparged and stirred bioreactor. The best biomass yield and root morphology were obtained in the air-sparged bioreactor with no stirring. The indole alkaloid contents of the roots cultivated in the bioreactor with stirring and in the bioreactor with medium circulation were significantly lower than that of the roots in the air-sparged bioreactor. Hairy roots of strawberry (Fragaria x ananassa Duch.) were cultivated in an air-sparged bioreactor with no stirring. The biomass yield was good and the roots were long and branching. The roots produced were used as a host for an endophyte, arbuscular mycorrhizal fungus, Glomus fistulosum V128.
    Original languageEnglish
    QualificationDoctor Degree
    Awarding Institution
    • University of Helsinki
    Supervisors/Advisors
    • Kauppinen, Veli, Advisor, External person
    Award date28 Oct 1994
    Place of PublicationEspoo
    Publisher
    Print ISBNs951-38-4637-7
    Publication statusPublished - 1994
    MoE publication typeG5 Doctoral dissertation (article)

    Fingerprint

    bioreactors
    cells
    air
    bubbles
    Catharanthus roseus
    embryo (plant)
    Fragaria ananassa
    Glomus
    biomass
    Betula
    pumps
    cell suspension culture
    strawberries
    mycorrhizal fungi
    cell culture
    experimental design
    cell lines
    indole alkaloids
    turbines

    Keywords

    • bioreactors
    • plant cell cultures
    • embryo cultures
    • suspension cultures
    • root hairs
    • Betula pendula
    • Catharanthus roseus
    • Fragaria x ananassa
    • root cultures

    Cite this

    @phdthesis{3645123b113349299b94cd407f13dcf6,
    title = "Plant cell and hairy root cultures in bioreactor-based production processes: Dissertation",
    abstract = "In this study, laboratory scale biotechnical production processes were developed for different types of plant cell cultures: embryogenic cell culture, undifferentiated cell suspension culture and hairy root culture. The production of somatic embryos, secondary metabolites and biomass were optimized. The growth as well as the production characteristics and the nutrient uptake of the different plant cell systems were determined. The suitabilities of different bioreactor configurations for the various culture types were tested in laboratory scale. In the optimization of growth and production, statistical experimental designs were used in addition to the conventional method of parallel testing of different media were used. The embryo production medium for an embryogenic cell line of birch (Betula pendula Roth) was optimized using a 22-factorial design. Factorial design was also tested for the optimization of alkaloid production medium for a cell suspension of Catharanthus roseus (L.) G. Don, but the conventional method of parallel testing of different media proved to be more successful. Factorial experimental design was used to determine the effects of low phosphate and sugar concentrations on strawberry (Fragaria x ananassa Duch.) hairy roots. In this way a slowly growing root system was obtained for infection studies with the native Finnish arbuscular mycorrhizal fungus, Glomus fistulosum V128. The embryogenic cell line of birch was cultivated in four different types of bioreactors: a sparged blade-stirred bioreactor, a diffusion-aerated bioreactor with a marine impeller (ChemCell, Chemap), an air-lift and a bubble column bioreactor. Embryo development was monitored during the cultivations by computer vision. The highest embryo production was achieved in the bubble column bioreactor. However, this was only 20{\%} of the production in control shake flasks. In the mechanically stirred bioreactors production was even lower, indicating shear damage of the embryos. The suspension culture of Catharanthus roseus L. was cultivated in three different types of bioreactors: a standard aerated and stirred bioreactor (control bioreactor), a recirculation bioreactor in which part of the air is recirculated with a peristaltic pump, and a bubble column bioreactor. The recirculation bioreactor was used to mimic the gaseous regime of a shake flask. When cultured in a turbine stirred tank reactor aerated through a sparger and in the bubble column the cells did not grow well and the production of ajmalicine was low. When cultivated in the recirculation bioreactor the biomass concentration and production of ajmalicine were similar to the level observed in shake flasks. Hairy roots of C. roseus were cultivated in three different types of bioreactors: an air-sparged bioreactor with no stirring, an air-sparged bioreactor in which the medium is slowly circulated through the vessel with a peristaltic pump and a standard air-sparged and stirred bioreactor. The best biomass yield and root morphology were obtained in the air-sparged bioreactor with no stirring. The indole alkaloid contents of the roots cultivated in the bioreactor with stirring and in the bioreactor with medium circulation were significantly lower than that of the roots in the air-sparged bioreactor. Hairy roots of strawberry (Fragaria x ananassa Duch.) were cultivated in an air-sparged bioreactor with no stirring. The biomass yield was good and the roots were long and branching. The roots produced were used as a host for an endophyte, arbuscular mycorrhizal fungus, Glomus fistulosum V128.",
    keywords = "bioreactors, plant cell cultures, embryo cultures, suspension cultures, root hairs, Betula pendula, Catharanthus roseus, Fragaria x ananassa, root cultures",
    author = "Anna-Maria Nuutila",
    note = "Project code: BIO9014 Project code: BIO9001 Project code: BEL2029 Project code: BEL4652",
    year = "1994",
    language = "English",
    isbn = "951-38-4637-7",
    series = "VTT Publications",
    publisher = "VTT Technical Research Centre of Finland",
    number = "199",
    address = "Finland",
    school = "University of Helsinki",

    }

    Plant cell and hairy root cultures in bioreactor-based production processes : Dissertation. / Nuutila, Anna-Maria.

    Espoo : VTT Technical Research Centre of Finland, 1994. 128 p.

    Research output: ThesisDissertationCollection of Articles

    TY - THES

    T1 - Plant cell and hairy root cultures in bioreactor-based production processes

    T2 - Dissertation

    AU - Nuutila, Anna-Maria

    N1 - Project code: BIO9014 Project code: BIO9001 Project code: BEL2029 Project code: BEL4652

    PY - 1994

    Y1 - 1994

    N2 - In this study, laboratory scale biotechnical production processes were developed for different types of plant cell cultures: embryogenic cell culture, undifferentiated cell suspension culture and hairy root culture. The production of somatic embryos, secondary metabolites and biomass were optimized. The growth as well as the production characteristics and the nutrient uptake of the different plant cell systems were determined. The suitabilities of different bioreactor configurations for the various culture types were tested in laboratory scale. In the optimization of growth and production, statistical experimental designs were used in addition to the conventional method of parallel testing of different media were used. The embryo production medium for an embryogenic cell line of birch (Betula pendula Roth) was optimized using a 22-factorial design. Factorial design was also tested for the optimization of alkaloid production medium for a cell suspension of Catharanthus roseus (L.) G. Don, but the conventional method of parallel testing of different media proved to be more successful. Factorial experimental design was used to determine the effects of low phosphate and sugar concentrations on strawberry (Fragaria x ananassa Duch.) hairy roots. In this way a slowly growing root system was obtained for infection studies with the native Finnish arbuscular mycorrhizal fungus, Glomus fistulosum V128. The embryogenic cell line of birch was cultivated in four different types of bioreactors: a sparged blade-stirred bioreactor, a diffusion-aerated bioreactor with a marine impeller (ChemCell, Chemap), an air-lift and a bubble column bioreactor. Embryo development was monitored during the cultivations by computer vision. The highest embryo production was achieved in the bubble column bioreactor. However, this was only 20% of the production in control shake flasks. In the mechanically stirred bioreactors production was even lower, indicating shear damage of the embryos. The suspension culture of Catharanthus roseus L. was cultivated in three different types of bioreactors: a standard aerated and stirred bioreactor (control bioreactor), a recirculation bioreactor in which part of the air is recirculated with a peristaltic pump, and a bubble column bioreactor. The recirculation bioreactor was used to mimic the gaseous regime of a shake flask. When cultured in a turbine stirred tank reactor aerated through a sparger and in the bubble column the cells did not grow well and the production of ajmalicine was low. When cultivated in the recirculation bioreactor the biomass concentration and production of ajmalicine were similar to the level observed in shake flasks. Hairy roots of C. roseus were cultivated in three different types of bioreactors: an air-sparged bioreactor with no stirring, an air-sparged bioreactor in which the medium is slowly circulated through the vessel with a peristaltic pump and a standard air-sparged and stirred bioreactor. The best biomass yield and root morphology were obtained in the air-sparged bioreactor with no stirring. The indole alkaloid contents of the roots cultivated in the bioreactor with stirring and in the bioreactor with medium circulation were significantly lower than that of the roots in the air-sparged bioreactor. Hairy roots of strawberry (Fragaria x ananassa Duch.) were cultivated in an air-sparged bioreactor with no stirring. The biomass yield was good and the roots were long and branching. The roots produced were used as a host for an endophyte, arbuscular mycorrhizal fungus, Glomus fistulosum V128.

    AB - In this study, laboratory scale biotechnical production processes were developed for different types of plant cell cultures: embryogenic cell culture, undifferentiated cell suspension culture and hairy root culture. The production of somatic embryos, secondary metabolites and biomass were optimized. The growth as well as the production characteristics and the nutrient uptake of the different plant cell systems were determined. The suitabilities of different bioreactor configurations for the various culture types were tested in laboratory scale. In the optimization of growth and production, statistical experimental designs were used in addition to the conventional method of parallel testing of different media were used. The embryo production medium for an embryogenic cell line of birch (Betula pendula Roth) was optimized using a 22-factorial design. Factorial design was also tested for the optimization of alkaloid production medium for a cell suspension of Catharanthus roseus (L.) G. Don, but the conventional method of parallel testing of different media proved to be more successful. Factorial experimental design was used to determine the effects of low phosphate and sugar concentrations on strawberry (Fragaria x ananassa Duch.) hairy roots. In this way a slowly growing root system was obtained for infection studies with the native Finnish arbuscular mycorrhizal fungus, Glomus fistulosum V128. The embryogenic cell line of birch was cultivated in four different types of bioreactors: a sparged blade-stirred bioreactor, a diffusion-aerated bioreactor with a marine impeller (ChemCell, Chemap), an air-lift and a bubble column bioreactor. Embryo development was monitored during the cultivations by computer vision. The highest embryo production was achieved in the bubble column bioreactor. However, this was only 20% of the production in control shake flasks. In the mechanically stirred bioreactors production was even lower, indicating shear damage of the embryos. The suspension culture of Catharanthus roseus L. was cultivated in three different types of bioreactors: a standard aerated and stirred bioreactor (control bioreactor), a recirculation bioreactor in which part of the air is recirculated with a peristaltic pump, and a bubble column bioreactor. The recirculation bioreactor was used to mimic the gaseous regime of a shake flask. When cultured in a turbine stirred tank reactor aerated through a sparger and in the bubble column the cells did not grow well and the production of ajmalicine was low. When cultivated in the recirculation bioreactor the biomass concentration and production of ajmalicine were similar to the level observed in shake flasks. Hairy roots of C. roseus were cultivated in three different types of bioreactors: an air-sparged bioreactor with no stirring, an air-sparged bioreactor in which the medium is slowly circulated through the vessel with a peristaltic pump and a standard air-sparged and stirred bioreactor. The best biomass yield and root morphology were obtained in the air-sparged bioreactor with no stirring. The indole alkaloid contents of the roots cultivated in the bioreactor with stirring and in the bioreactor with medium circulation were significantly lower than that of the roots in the air-sparged bioreactor. Hairy roots of strawberry (Fragaria x ananassa Duch.) were cultivated in an air-sparged bioreactor with no stirring. The biomass yield was good and the roots were long and branching. The roots produced were used as a host for an endophyte, arbuscular mycorrhizal fungus, Glomus fistulosum V128.

    KW - bioreactors

    KW - plant cell cultures

    KW - embryo cultures

    KW - suspension cultures

    KW - root hairs

    KW - Betula pendula

    KW - Catharanthus roseus

    KW - Fragaria x ananassa

    KW - root cultures

    M3 - Dissertation

    SN - 951-38-4637-7

    T3 - VTT Publications

    PB - VTT Technical Research Centre of Finland

    CY - Espoo

    ER -