PREPARATION OF METABOLICALLY ACTIVE PROTOPLASTS OF BLUE-GREEN ALGAE(1)

Area of Science:

• Microbiology
• Phycology
• Biochemistry

Background:

• Cyanobacteria, including Microcystis aeruginosa, Anacystis nidulans, and Anabaena flos-aquae, are vital primary producers in aquatic ecosystems.
• Protoplast isolation is crucial for genetic manipulation and physiological studies of cyanobacteria.
• Previous methods for protoplast isolation from these species were limited by cell viability and metabolic activity.

Purpose of the Study:

• To develop a method for isolating metabolically active protoplasts from key cyanobacterial species.
• To assess the physiological integrity (photosynthesis and respiration) of the isolated protoplasts.
• To identify potential lysozyme-resistant mutants for further research.

Main Methods:

• Treatment of Microcystis aeruginosa, Anacystis nidulans, and Anabaena flos-aquae cells with lysozyme.
• Culturing of isolated protoplasts to assess viability and metabolic activity over several days.
• Measurement of photosynthetic and respiratory rates in protoplasts compared to intact control cells.
• Isolation and characterization of lysozyme-resistant cell populations.

Main Results:

• Metabolically active protoplasts were successfully obtained from the studied cyanobacteria.
• Isolated protoplasts maintained photosynthetic and respiratory rates comparable to those of intact control cells.
• Lysozyme-resistant cells were identified, suggesting the potential for genetic variation or mutation.

Conclusions:

• Lysozyme treatment is an effective method for producing metabolically active protoplasts from Microcystis aeruginosa, Anacystis nidulans, and Anabaena flos-aquae.
• The isolated protoplasts retain significant physiological functions, making them suitable for further studies.
• The discovery of lysozyme-resistant strains opens avenues for investigating genetic diversity and developing novel strains.