Optimization of Protoplast Isolation Protocols from Callus Culture of Anacardium occidentale L.

摘要

Protoplast technology endeavors a unique single cell system that promotes several aspects of modern biotechnology. In the present study, an efficient protocol to isolate the protoplast from callus culture of a valuable plantation crop, Anacardium occidentale was developed. The phenolic compound released during digestion process caused protoplast injury resulted in low yielding. Further investigations revealed that pH 5.0 and osmotic concentration 0.5M mannitol were found suitable for maximum protoplasts yield. The results also indicated that the age of the callus tissues plays a key role in the yield of protoplasts in A.occidentale. During the study, youngcalli showed maximum protoplast yield when treated with cellulase (2.5 %), pectinase (2.5 %), hemicellulase (1.5%) and 20mM CaCl2 in 0.5M mannitol. Meanwhile, an incubationperiod of 4hours with enzyme solution resulted in the maximum yield of protoplasts (4.71 ± 0.15)xl03 protoplasts/g FW. In short, the results generated in the study can be usedas a platform to work out the protoplast technology in A.occidentale.