Immobilization of plant protoplasts: Viability studies.

Area of Science:

• Plant biotechnology
• Cell immobilization techniques
• Biopolymer applications

Background:

• Plant protoplast culture is vital for genetic engineering and cell biology.
• Protoplasts are sensitive to environmental stresses, limiting their long-term survival.
• Developing robust methods for protoplast stabilization is crucial for biotechnological applications.

Purpose of the Study:

• To immobilize Daucus carota and Catharanthus roseus protoplasts using gel-forming polysaccharides.
• To evaluate the viability and stability of immobilized protoplasts compared to free cells.
• To explore the potential of polysaccharide matrices for enhancing protoplast culture.

Main Methods:

• Protoplast isolation from Daucus carota Ca68 and Catharanthus roseus.
• Immobilization via entrapment in kappa-carrageenan, agarose, and alginate gels.
• Preparation of uniform spherical beads using an inert hydrophobic phase (vegetable oil).
• Assessment of protoplast viability and integrity using standard assays and enzyme activity (digitoxigenin 58-hydroxylase).

Main Results:

• Immobilized protoplasts in carrageenan and agarose beads demonstrated enhanced viability after 14 days compared to free protoplasts.
• The polysaccharide matrix provided stabilization against osmotic shock.
• Viability monitoring using digitoxigenin 58-hydroxylase activity confirmed the stability of Daucus carota protoplasts.

Conclusions:

• Entrapment in polysaccharide gels (carrageenan, agarose) is an effective method for immobilizing and stabilizing plant protoplasts.
• Immobilization significantly improves protoplast viability under culture conditions.
• This technique offers a promising approach for long-term plant cell culture and biotechnological applications.