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  • Protoplast: Norazian Mohd Alwi MQ082019

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    Brintha Ganapathy
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    Protoplasts are plant, bacterial, or fungal cells that have had their cell walls removed, either mechanically or enzymatically. They appear spherical and float freely in solution. The sizes of protoplasts from the same source are generally consistent. Cell walls can be removed from plant cells using cellulase, pectinase and xylanase enzymes. Lysozyme plus EDTA is used to remove the cell walls of gram-positive bacteria, while chitinase is used for fungal cells. Lysozyme catalyzes the breakdown of peptidoglycan in gram-positive bacterial cell walls. Protoplasts are useful for applications like transformation and protoplast fusion for plant breeding.

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    Lecture 4 Pro Top Lasts

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    Protoplasts are plant, bacterial, or fungal cells that have had their cell walls removed, either mechanically or enzymatically. They appear spherical and float freely in solution. The sizes of protoplasts from the same source are generally consistent. Cell walls can be removed from plant cells using cellulase, pectinase and xylanase enzymes. Lysozyme plus EDTA is used to remove the cell walls of gram-positive bacteria, while chitinase is used for fungal cells. Lysozyme catalyzes the breakdown of peptidoglycan in gram-positive bacterial cell walls. Protoplasts are useful for applications like transformation and protoplast fusion for plant breeding.

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    20 views15 pages

    Protoplast: Norazian Mohd Alwi MQ082019

    Uploaded by

    Brintha Ganapathy
    Protoplasts are plant, bacterial, or fungal cells that have had their cell walls removed, either mechanically or enzymatically. They appear spherical and float freely in solution. The sizes of protoplasts from the same source are generally consistent. Cell walls can be removed from plant cells using cellulase, pectinase and xylanase enzymes. Lysozyme plus EDTA is used to remove the cell walls of gram-positive bacteria, while chitinase is used for fungal cells. Lysozyme catalyzes the breakdown of peptidoglycan in gram-positive bacterial cell walls. Protoplasts are useful for applications like transformation and protoplast fusion for plant breeding.

    Copyright:

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    of 15

    PROTOPLAST

    NORAZIAN MOHD ALWI MQ082019

    Definition
    Protoplast: is a plant, bacterial or fungal cell that had its cell wall removed. using either mechanical or enzymatic means.

    Appearance
    spherical, clearly 3-dimensional float freely within the solution

    The sizes of the protoplasts are generally consistent

    Protoplast

    Fusion protoplast

    Protoplast

    Degrading Enzymes
    Type of cell Plant cells Enzyme Cellulase, pectinase, xylanase Lysozyme (+EDTA)

    Gram-positive bacteria
    Fungal cells

    Chitinase

    LYSOZYME
    Occurs naturally in some eukaryotes cells. Found in tears, mucous, saliva and tissue fluid. It exhibits its antimicrobial activity.

    LYSOZYME (cont.)
    Active on the major cell wall component of gram-positive, making them vulnerable to lysis. Catalyzes hydrolysis of the bond between the sugars in the repeating disaccharides backbone of peptidoglycan.

    Cell Walls : Peptidoglycan

    Protoplasts

    The gram-positive cell wall is almost completely destroyed by lysozyme.

    Spheroplasts The gram-negative cell wall is not destroyed by lysozyme same extent as in gram positive cells. Requires pretreatment to destabilize outer membrane. Treated with EDTA. (weaken ionic bonds in the outer membrane and damage it)

    Cell wall of Gram negative vs positive

    During and subsequent digestion of the cell wall, the protoplast becomes very sensitive to osmotic stress.

    This means cell wall digestion and protoplast storage must be done in an isotonic solution to prevent rupture of the plasma membrane.

    Application
    To study membrane biology, including the uptake of macromolecules and viruses.
    Used for DNA transformation (for making genetically modified organisms), since the cell wall would otherwise block the passage of DNA into the cell.

    Applications (cont.)
    Used for plant breeding, using a technique called protoplast fusion. Protoplasts from different species are induced to fuse by using an electric field or a solution of polyethylene glycol. This technique may be used to generate somatic hybrids in tissue culture.

    The end..

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