Genetic Modification Transformation of Protoplasts, Protoplasts Lack a Cell Wall, Around Plasma Membrane, Chloroplasts, Nuclei, Microorganisms

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Protoplast Culture Regeneration and Somatic Hybridization Protoplast Culture Regeneration and Somatic Hybridization - Introduction Isolation of Proplasts Mechanical Method Enzymatic Method Direct One Step Method Sequential Two Step Method Purification of Isolated Protoplasts Sedimentation and Washing Flotation Viability and Plating Density of Protoplasts Testing Protoplast Viability Minimum Plating Density MPD and Culturing Individual Protaplasts Protaplast Culture and Regeneration of Plants Isolation of Subprotoplasts Protoplast Fusion and Somatic Hybridization Techniques of Protoplast Fusion

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Genetic Modification (Transformation) of Protoplasts

Since protoplasts lack a cell wall around its plasma membrane, they are suitable for uptake of chloroplasts, nuclei, microorganisms and naked DNA.

Although, some success has been achieved for the introduction of foreign chloroplasts, nuclei and microbes for genetic modification, the maximum work has been done and success achieved in the uptake of foreign DNA.

The genetic Modification through uptake of foreign DNA involves uptake, integration, replication, expression and transmission. This will be discussed in greater detail in the next chapter.

A Diagrammatic Representation of the wide Range of Biological Components

A Diagrammatic Rerpesentation of the wide range of biological components that can be takens up by isolated protoplasts

1. Chromosome	4. plasmids or naked DNA
2. mitochondria	5. nuclei
3. chloroplast	6. bacteria

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Induced Fusion NaNo₃-treatment Treatment with Calcium Ions ca⁺⁺ at High pH Polyethylene Glycol PEG Treatment Electrical Fusion Selection of Fused Protaplasts Chromosome Status of Somatic Hybrids Interspecific Hybrids Produced Through Protoplast Fusion Intergeneric Hybrids Produced Through Protaplast Fusion Use of Protoplast Fusion and Somatic Hybrids Somatic Hybrids for Gene Transfer Asymmetry in Somatic Hybrids for Gene Transfer Somatic Hybrids for Cytoplasmic Male Sterility Cytoplasmic Hybrids or Cybrids Genetic Modification Transformation of Protoplasts