Techniques of Protoplast Fusion, Spontaneous Fusion, Protoplasts for Culture, Enzymatic Degradation, Walls is Affected, Homokaryons or Homokaryocytes

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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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Techniques of protoplast fusion

Spontaneous fusion.

During isolation of protoplasts for culture, when enzymatic degradation of cell walls is affected, some of the protoplasts, lying in close proximity, may undergo fusion to produce homokaryons or homokaryocytes, each with 2-40 nuclei.

The occurrence of multinucleate fusion bodies is more frequent, when protoplasts are prepared from actively dividing cells.

This spontaneous fusion however, is strictly intraspecific. However spontaneous fusion of protoplasts can also be induced by bringing protoplasts into intimate contact through micromanipulators or micropipettes.

There seems to be a correlation between the age of the leaf used for isolation of protoplasts and the percentage of protoplasts undergoing spontaneous fusion; protoplasts from young leaves are more likely to undergo this fusion.

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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

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