Isolation of Subprotoplasts, Cytoplasts Lack Nuclei, Karyoplasts, Miniprotoplasts, Nucleus Surrounded, Plasma Membrane, Laser Beam Microsurgery

Scholarships Dictionary Visa Details Study Abroad Exam Results Online Test Jobs

	Home
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

Home >> Plant Biotechnology and Genomics >> Protoplast Culture Regeneration and Somatic Hybridization >> Isolation of Subprotoplasts

Isolation of Subprotoplasts

Subprotoplasts, which do not contain each the entire content of a plant cell, include the following four types:

(i) cytoplasts lack nuclei and each cytoplast contains entire cytoplasm of a cell or a part thereof; these are often used for cybrid production through somatic hybridization (consult next section), which may sometimes lead to the transfer of a part of the genome;

(ii) karyoplasts or miniprotoplasts contain, each a nucleus surrounded by some cytoplasm and the original outer plasma membrane;

(iii) microprotoplasts contain each, only a few of all chromosomes, and a fraction of the cytoplasm;

(iv) microplasts contain each, only a fraction of cytoplasm and outer membrane.

Subprotoplasts can be prepared by one of the following methods:

(i) Cytoplasts can be obtained by X-ray treatment or laser beam microsurgery of freshly isolated protoplasts; the nucleus is inactivated by such treatment.

(ii) Cytoplasts occur spontaneously in the pericarps of ripening fruits as shown in tomato; these cytoplasts can be isolated by filtration of the juice through sieves followed by sedimentation and centrifugation.

(iii) Enucleated protoplasts may also be obtained from cultured protoplasts due to phenomenon of ‘budding’ (shaking of protoplasts may lead to separation of budded Subprotoplasts); these may sometimes represent microprotoplasts.

(iv) Cytoplasts and miniprotoplasts are also obtained through plasmolysis of elongated cells.

When Plasmolysed Cells are Enzyme Treated

When Plasmlysd cell are enzyme treated they will often yield a mixture of protoplasts (pp), miniprotoplasts and cytoplasts

	Home
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