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AN EFFICIENT METHOD OF PROTOPLAST ISOLATION IN BANANA (MUSA SPP.)

ABDULLAH KHATRI¹, MUHAMMAD UMAR DAHOT², IMTIAZ AHMED KHAN¹ AND GHULAM SHAH NIZAMANI¹

Abstract: Protoplast cultures are essential commodity for transformation and this can be achieved through electroporation and somatic hybridization. Like other monocot systems, embryogenic cell suspension is the material of choice for the isolation of protoplasts. In banana protoplasts have been successfully isolated but so far sufficient quantities of protoplasts for practical application are not routinely met. Enzyme mixture comprises of 3% cellulose R-10, 1% macerozyme R-10 and 1% pectinase was used for protoplast isolation. Leaf, leaf bases and corm tissue were used as an explant source and corm tissue gave the highest yields of protoplasts. Average yield of protoplasts from corm tissue ranged from 2.5 x 10⁵ to 8.7 x 10⁵ protoplasts. Freshly isolated protoplasts were colorless and 80% protoplasts stained with FDA shows high level of viability. Protoplast were cultured for callus induction on SH medium  and cell wall formation was observed after 3 days of culture and cell division was achieved within 6-8 days.

¹Nuclear Institute of Agriculture, Tando Jam

²Institute of Biotechnology and Genetic Engineering, University of Sindh, Jamshoro