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    Форум Cайта » Plants & Life » Plants » Plant in vitro Culture and Propagation in Agar
    Plant in vitro Culture and Propagation in Agar
    Syler_91Date: Sunday, 09.11.2014, 00:43 | Message # 1
    Group: Администраторы
    Messages: 99
    Reputation: 2
    Status: Offline
    A procedure to regenerate plants genetically identical to the starting one.
    Agar, agarose and alginates can be used as a
    solid support for the growth of cells of bacteria, animals or plants. In
    fact, the cells and the tissues of many organisms are able to multiply
    and to form new organisms, if they are grown in the proper environment
    in presence of all the needed nutrients. The "in vitro" culture
    technology of plant cells is simpler than that of animal cells. A
    culture medium containing all the nutritive elements (salts and
    vitamins, sugars) must be prepared and dispensed in jars where to insert
    a plant tissue explants. The culture is then maintained in proper
    conditions of light and temperature that vary from one case to another.
    Within days/weeks the growth of undifferentiated calli or differentiated
    tissues (buds, roots, etc) will be observed. It is possible with this
    procedure to regenerate the whole plant, genetically identical to the
    starting one.

    Material: Glass jars, Plant tissues, Culture medium, Agar powder, Sucrose powder, Autoclave, Bleach, Beakers, and Bunsen burner.

    Prepare the plant culture medium. It is convenient to
    use a commercial formula of lyophilized Murashige and Skoog medium to
    which to simply add water and sucrose (3g/liter of medium) as carbon
    source. Add agar (0.5-0.8%) to the medium. Sterilize the utensils the
    agar-medium, and the jars putting them for a 21 minutes cycle in
    autoclave or in a pressure cooker. Let the medium cool till 45-50°C and
    dispense the medium in the jars when the agar is still liquid. Sterilize
    the plant tissue explants incubating them in beakers using diluted
    (1/10) commercial bleach (sodium ipoclorite, 40% solution) for 15
    minutes. When the agar in the jar has solidified insert 2-3 plant tissue
    explants. All the procedures must be done under sterile conditions. If
    possible use a sterile air cabinet or operate on a clean bench, far from
    wind and dust, always working near to a Bunsen burner, so that the area
    could stay aseptic. In laboratories for in vitro plant propagation, as
    those of the CNR-IGV in Portici, tissue cultures are maintained in
    growth chambers at controlled light and temperature conditions.
    Форум Cайта » Plants & Life » Plants » Plant in vitro Culture and Propagation in Agar
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