Protoplast Culture Regeneration and Somatic Hybridization Introduction, Significant Developments, Tissue Culture, Plasma Membrane

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Protoplast Culture Regeneration and Somatic Hybridization Protoplast Culture Regeneration and Somatic Hybridization - Introduction Isolation of Proplasts Isolation of Proplasts - Mechanical Method Isolation of Proplasts - Enzymatic Method Isolation of Proplasts - Direct One Step Method Isolation of Proplasts - 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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Protoplast Culture, Regeneration and Somatic Hybridization

One of the most significant developments in the field of plant tissue culture, witness during the last few decades, is the isolation, culture and fusion of protoplasts. A more recent achievement is the manipulation and regeneration of these cultured or fused protoplasts into whole plants.

Since in plant cells, the plasma membrane in bound by a rigid cellulose wall (unlike animal cells), it has been relatively difficult to handle plant cells. Only in 1960, it was demonstrated by E. C. Cocking at the University of Nottingham (U.K.), that naked cells calls protoplasts can be obtained through enzymatic degradation of cell walls.

This led to significant developments in the field of somatic cell genetics in higher plants. Cultured protoplasts can be used not only for somatic cell fusions, but also for taking up foreign DNA, cell organelles, bacteria and virus particles.

In view of this, the isolation and culture of protoplasts has become a very important area of research, within the realm of plant biotechnology.

The essential ingredients of the technique include isolation of protoplasts, culture of protoplasts, introduction of foreign DNA into protoplasts, raising whole plants from cultured protoplasts and fusion of protoplasts leading to somatic hybridization. These aspects of plant tissue culture will be discussed in this.

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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