Vol. 9, Issue 7, Part U (2025)

This study establishes a reliable and efficient protocol for protoplast isolation from leaf explants of soybean (Glycine max L.) cv. MAUS 158, a stem fly-tolerant and high-yielding genotype. leaf tissue was utilized to evaluate explant-specific enzymatic digestion conditions. Preplasmolysis was performed using 13% mannitol, followed by enzymatic digestion with a combination of 2% cellulase and 1% macerozyme. Optimal incubation times 18 was determined for leaf tissue. The protoplast was purified using a 21% sucrose gradient and assessed for yield and viability. Leaf tissue showed the highest protoplast yield (3.60 × 10⁴ cells/g), highest viability rates (>83%). The study confirms that the selected enzyme combination, in conjunction with explant-specific digestion times, supports the successful isolation of viable protoplast from soybean leaf tissues. This optimized protocol is applicable for advanced studies in plant regeneration, somatic hybridization, and genetic transformation.