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Pak. J. Bot., 47(6): 2391-2396, 2015.

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A STUDY ON THE ISOLATION OF PROTOPLASTS FROM THE CALLUS OF LILIUM LONGIFLORUM OVERIG

SARAH YOUSUF¹, FARYAL ASHRAF¹, SYEDA KAHKASHAN KAZMI², SAIFULLAH KHAN^{2, 3*}AND HAMMAD AFZAL KAYANI^2,4

¹Department of Biotechnology, Faculty of Science, University of Karachi, Karachi-75270, Pakistan

²Biotechnology Wing, H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences (ICCBS), University of Karachi-75270, Pakistan

³Department of Agriculture and Agribusiness Management, University of Karachi, Pakistan

⁴Shaheed Zulfiqar Ali Bhutto Institute of Science and Technology, Karachi, Pakistan

^*Corresponding author’s e-mail: drsaif65@gmail.com

Abstract: Lilium longiflorum Overig is a Lilaceous plant grown for ornamental as well as certain other general purposes. The research presented here focuses on the method of protoplast isolation from the In vitro grown callus of Lilium longiflorum Overig. Series of experiments were conducted in order to optimize the conditions. Four different amounts of callus and a range of incubation time were used to achieve maximum number of viable protoplasts. However, the calli used were treated with Enzyme solution containing 0.5% (w/v) Macerozyme, 2% (w/v) Cellulase and 0.1% (w/v) Pectinase in combination, to obtain maximum number of viable protoplasts through the process of cell-wall digestion. Furthermore, an osmoticum of 20% (w/v) sucrose and a washing solution consisting of 0.1% (w/v) MES and 0.5M Mannitol were added to suspend a ring of protoplast at the interphase of both the solutions. This ring of protoplast was then isolated and tested for viability and yield. Consequently, amongst the various sets of experiment, callus of 1.5 grams and incubation time of 4 hours was found an ideal approach to obtain maximum number of viable protoplasts.

Key words: Protoplast isolation, Easter lily, Protoplast yield, Protoplast viability, Lily calli.