A SIMPLE METHOD FOR THE ISOLATION OF PROTOPLASTS FROM LEAVES OF NICOTIANA BENTHAMIANA
K. M. K. Huda* and M. A. Siddikee
*Corresponding author: Email mashnuk@gmail.com
Abstract
Protoplasts are plant cells that lack their cell walls. Isolation of protoplasts from different plant tissues was first reported more than 40 years ago and since then used to study a variety of cellular processes. In this article, a simple protoplast isolation method was developed from the leaves of Nicotiana benthamiana grown in the open air. The optimum condition for protoplast isolation was established by using 1% cellulose R-10 and 0.3% macerozyme R-10 dissolved in F-PIN solution. The method is simple and easy-to-handle and gives satisfactory yields typically. Two to three leaves of tobacco yielded 5-10x106 intact protoplasts in 2 hr.
Keywords: Isolation; leaves; protoplasts; simple protocol; tobacco
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