A simple protocol for a rapid and consistent production of a large number of viable protoplasts from the Ulvophycean species

Protoplasts are extensively employed in plant cellular biology to develop somatic hybrids with agronomically important traits. The success of cellular biotechnology for marine macroalgae is in its inception owing to lack of a simple unified protocol for protoplasts preparation. Previous studies have employed multiple enzymes, mostly crude enzymes prepared in the laboratory from either microbes or marine animals that were invariably a source of inconsistent results. This study reports a simple protocol with reproducible protoplast yields from Ulvophycean species, and the advantage includes the use of a single commercial enzyme at a much lower concentration in low ionic strength medium. In addition, the enzyme reusability was established five times for the first time without compromising on the protoplast yield and viability. Development of such a facile protocol will propel the application of protoplasts in cellular biotechnology of Ulvophyceaen species, which have nowadays gained considerable importance in food and energy perspective. Keywords: Enzyme, genetic modification, green seaweed, protoplast, Ulva