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#protoplast fusion

12 public questions tagged with this topic.

Plant tissue culture 2

Fusion of cytoplasm of two cells with nucleus from one parent is called:

Fusion product containing cytoplasm of two cells but nucleus from only one parent is termed cybrid cytoplasmic hybrid, distinction from true somatic hybrid which contains both nuclei yielding allotetraploid. Cybrids arise when protoplasts of two sources fuse forming heterokaryon with two nuclei and mixed organelles followed by selective loss or inactivation of one nucleus. Inactivation can be engineered by pre treating one partner with gamma irradiation X ray or metabolic inhibitor iodoacetate that damages nucleus while preserving cytoplasm viability. During subsequent mitosis heterokaryon res

Ref: Sidorov et al., 1981 cybrid concept; Gleba 1979 Cytoplasmic inheritance.

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Plant tissue culture 2

Electrofusion induces fusion by:

Electrofusion induces protoplast fusion by applying controlled electrical pulses that create reversible membrane breakdown at points of contact allowing lipid bilayers to merge and cytoplasms to coalesce, offering higher fusion frequency viability and selectivity than chemical PEG method. Protocol consists of two electrical phases: first low strength high frequency alternating current 50 to 200 kilohertz field causes dielectrophoresis polarizing protoplasts forming induced dipoles aligning them in pearl chains bringing membranes into intimate contact at poles. Second short high intensity direc

Ref: Zimmermann & Vienken 1982 electrofusion; Tempelaar et al., Plant Sci electroporation.

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Plant tissue culture 2

Common fusogen used for induced protoplast fusion is:

Common chemical fusogen employed to induce protoplast fusion is polyethylene glycol PEG polymer with molecular weight typically 1500 to 6000 daltons applied at concentration 15 to 40 percent weight per volume combined with calcium ions 50 millimolar and alkaline pH 9 to 10. PEG induces fusion through multiple mechanisms: it is highly hydrophilic binding water molecules dehydrating intermembrane space reducing hydration repulsion between negatively charged phospholipid bilayers, promotes close apposition of membranes via calcium bridging between phosphate head groups and creates molecular disor

Ref: Kao & Michayluk 1974 PEG fusion; NCBI protoplast fusion review.

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Plant tissue culture 2

Somatic hybridization involves fusion of:

Somatic hybridization involves fusion of protoplasts isolated from somatic vegetative tissues of different species cultivars or genera, bypassing sexual incompatibility barriers that prevent conventional crossing due to pre zygotic pollen stigma incongruence or post zygotic endosperm abortion. Protoplasts are obtained by enzymatic digestion of cell walls using cellulase Onozuka R-10 and macerozyme releasing naked cells bounded solely by plasma membrane maintaining totipotency to regenerate wall and divide. Mixed protoplast populations labeled with different fluorescent markers or selectable co

Ref: Gleba & Sytnik 1984 somatic hybridization; Evans et al., Handbook Plant Cell Culture.

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