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    Mun Hui Sin Nuraini Binti Abdullah Siti Rabaah Binti Hamzah

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    This document describes two methods of plant cell culture: callus culture and suspension culture. Callus culture involves placing plant explants on a solid medium to produce an unorganized cell mass, while suspension culture transfers friable callus cells to a liquid medium where they detach and grow independently. Both methods use auxins and cytokinins to promote cell growth and require subculturing cells periodically into fresh media to maintain viability.

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    Mun Hui Sin Nuraini Binti Abdullah Siti Rabaah Binti Hamzah

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    TANSIEWYEN
    65 views15 pages
    This document describes two methods of plant cell culture: callus culture and suspension culture. Callus culture involves placing plant explants on a solid medium to produce an unorganized cell mass, while suspension culture transfers friable callus cells to a liquid medium where they detach and grow independently. Both methods use auxins and cytokinins to promote cell growth and require subculturing cells periodically into fresh media to maintain viability.

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    Method of Plant Cell Culture

    Original Title

    Plant Culture

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    This document describes two methods of plant cell culture: callus culture and suspension culture. Callus culture involves placing plant explants on a solid medium to produce an unorganized cell mass, while suspension culture transfers friable callus cells to a liquid medium where they detach and grow independently. Both methods use auxins and cytokinins to promote cell growth and require subculturing cells periodically into fresh media to maintain viability.

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    © All Rights Reserved
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    Download as pptx, pdf, or txt
    65 views15 pages

    Mun Hui Sin Nuraini Binti Abdullah Siti Rabaah Binti Hamzah

    Uploaded by

    TANSIEWYEN
    This document describes two methods of plant cell culture: callus culture and suspension culture. Callus culture involves placing plant explants on a solid medium to produce an unorganized cell mass, while suspension culture transfers friable callus cells to a liquid medium where they detach and grow independently. Both methods use auxins and cytokinins to promote cell growth and require subculturing cells periodically into fresh media to maintain viability.

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    Mun Hui Sin

    Nuraini Binti Abdullah


    Siti Rabaah Binti Hamzah
    Tan Siew Yen

    Method 1 - Callus Culture

    Callus is explants that cultured on the appropriate medium,


    with auxin and a cytokinin, give rise to an unorganised,
    growing and dividing mass of cells.

    Any plant tissue that have dividing cells can be used as an


    explant.

    Exercised plant material is placed on solidified medium


    containing nutrients and hormones that promote rapid cell
    differentiation.

    Callus forms can be quite large (>1cm) (Shuler and Kargi,


    2002).

    Method 1 - Callus culture

    During callus formation there is some degree of


    dedifferentiation or changes in morphology and metabolism
    that can cause plant cultures to lose the ability to
    photosynthesise.

    Overcome by the addition of other components such as


    vitamins and carbon source to the culture medium.

    Method 1 - Callus culture

    Figure 1: Function of Auxin and Cytokinin

    Auxin Promote root initiation.


    Cytokinin Induce growth of shoots

    Method 2 - Suspension Culture


    There are two categories of callus cultures: compact or
    friable. In compact callus, the cells are densely aggregated
    while in friable callus the cells are only loosely associated
    with each other and the callus becomes soft and breaks apart
    easily.
    Friable callus provides the inoculum to form cell-suspension
    cultures.
    The friability of callus is improved by manipulating the
    medium components, repeated subculturing or culturing it on
    medium with a low concentration of gelling agent.

    Method 2 - Suspension Culture

    Callus culture is then transferred into liquid medium to


    establish suspension culture, which is place on shaker to
    supply the cells with sufficient oxygen.

    With moderate agitation, cells or small aggregates of cells


    will slough off.

    A platform shaker is used to give a circular motion in a


    viable speed control (30-150 rpm) (Shuler and Kargi, 2002).

    Callus culture is performed in the dark (27C & pH 5.5) as


    light can encourage differentiation of the callus (Shuler and
    Kargi, 2002).

    Method 2 - Suspension Culture

    During long-term culture, habituation occurs. The culture may


    lose the requirement for auxin and/or cytokinin.

    Under the correct conditions, these released cells continue to


    grow and divide, eventually producing a cell-suspension culture.
    The inoculum should not be too large as toxic products released
    from damaged or stressed cells can build up to lethal levels.
    Cell suspensions can be maintained by batch cultures in conical
    flasks. They are continually cultured by repeated subculturing
    into fresh medium.

    Method 2 - Suspension Culture


    This results in dilution of the suspension and the initiation of
    another batch growth cycle. The degree of dilution during
    subculture should be determined empirically for each culture.
    Too great a degree of dilution will result in a greatly extended
    lag period or, in extreme cases, death of the transferred cells.
    After subculture, the cells divide and the biomass of the culture
    increases, until nutrients in the medium are exhausted and/or
    toxic by-products build up to inhibitory (stationary phase).
    If cells are left in the stationary phase for too long (~2-3 weeks),
    they will die and the culture will be lost. Therefore, cells should
    be transferred to fresh medium as they enter the stationary phase
    (Shuler and Kargi, 2002).

    Figure 2: Steps in Plant Cell Culture (Dodds & Roberts, 1995).

    Figure 3: Steps in Plant Cell Culture (Shuler and Kargi, 2002).

    Figure 4: Callus Culture (right) and Suspension Culture (left) of


    Coleus Blumei (Dodds & Roberts, 1995).

    References
    Dodds, J. H. and Roberts, L. W. (1995). Experiments in plant tissue
    culture. Cambridge University Press, Cambridge.
    Shuler, M. L. and Kargi, F. (2002). Bioprocess Engineering: Basic
    Concept. 2nd Ed. Upper Saddle River, NJ: Prentice Hall PTR

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