Treatment with Calcium Ions Ca++ at High pH, Involves Spinning, Centrifugation the Protoplasts, Calcium Cloride, Fusion Inducing Solution

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Home >> Plant Biotechnology and Genomics >> Protoplast Culture Regeneration and Somatic Hybridization >> Treatment with Calcium Ions Ca++ at High pH

Treatment with calcium ions (Ca⁺⁺) at high pH.

This method involves spinning (centrifugation the protoplasts in a fusion inducing solution (0.05M CaCl₂ 2H₂O in 0.4M mannitol at pH 10.5) for 30 minutes at 50g, after which the tubes are placed in a water bath (37°C) for 40-50 minutes.

This leads to fusion of 20-50% of the protoplasts. The details of the protocol are described by Bhojwani and Razdan (1983). The method was found to be superior to other methods in some cases, but high pH was toxic in other cases.

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