Protoplast Isolation and Regeneration from Gracilaria changii (Gracilariales, Rhodophyta)

Yeong, H.Y. and Khalid, N. and Phang, S.M. (2008) Protoplast Isolation and Regeneration from Gracilaria changii (Gracilariales, Rhodophyta). Journal of Applied Phycology, 20 (5). pp. 641-651. ISSN 09218971

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Abstract

The tropical agarophyte Gracilaria changii has been much researched and documented by the Algae Research Laboratory, University of Malaya, especially with regards to its potential as a seaweed bioreactor for valuable compounds. Protoplast regeneration of this seaweed was developed following the optimization of protoplast isolation protocol. Effect of the concentration and combination of isolating enzymes, incubation period, temperature, enzyme solution pH, tissue source on the protoplast yields were used to optimize the isolation protocol. The enzyme mixture with 4 w/v cellulase Onozuka R-10, 2 w/v macerozyme R-10 and 1 unit mL-1 agarase was found to produce the highest yield of protoplast at 28°C and 3 h incubation period. Thallus tips gave higher yields of protoplasts than middle segments. Freshly isolated G. changii protoplasts were cultured in MES medium. Regeneration of protoplast cell walls after 24 h was confirmed by calcofluor white M2R staining under UV fluorescence microscopy. The protoplasts with regenerated cell walls then underwent a series of cell division to produce callus-like cell masses in MES medium. Following this, juvenile plants of G. changii were obtained.

Item Type: Article
Uncontrolled Keywords: Callus Cell wall degrading enzymes Gracilaria changii Protoplast isolation Protoplast regeneration algae Gracilaria Gracilariales Rhodophyta
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: Faculty of Science > Institute of Biological Sciences
Depositing User: miss munirah saadom
Date Deposited: 08 May 2013 01:31
Last Modified: 08 May 2013 01:31
URI: http://eprints.um.edu.my/id/eprint/6006

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