EFFECT OF IODOACETAMIDE (IOA) ON THE VIABILITY AND CELL DIVISION OF CULTURED PROTOPLASTS OF MUNGBEAN (Vigna radiata (L.) Wilczek)

• Main Author: (NIM 206 97 502), Balqis
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/7383
• Institution: Institut Teknologi Bandung

Abstract : <br /> <br /> <br /> The effect of iodoacetamide (IOA) on the viability and cell divison of cultured protoplasts of mungbean (Vigna radiata (L.) Wilczek) var. Walet have been studied. IOA is metabolic inhibitor that inactivate cell cytoplasm and inhibit cell division. The aim of this research was to evaluate the effect of IOA on viability and IOA concentration which inhibit cell division of mungbean protoplasts. The concentration of IOA used were 2.5; 5.0; 7.5; 10.0; and 12.5 mM. Protoplasts were isolated enzymatically from the first leaves of 7 days old seedling and hypocotyl of 5 days old seedling. The enzyme solution consisted of 1.5% Cellulase RS, 1% Macerozyme R-10, 0.4 M mannitol and 2.5 mM CaC12.2H20. The Protoplasts were immobilized in alginate beads and cultured in KM-8p medium. The protoplasts density were 5.105/ml and 105/ml, from leaf and hypocotyl respectively. The protoplast viability was observed using fluorescence microscope following FDA staining. Cell wall regeneration was observed using fluorescence microscope following Calcofluor White staining. Cell division and colony formation were observed using inverted microscope. The result showed that 12.5 mM IOA decreased protoplasts viability, especially leaf protoplasts. All of the IOA concentration used inhibited cell wall regeneration, indicated by longer periode of regeneration of cell wall. IOA also inhibited cell division. There were no cell division occurred on leaf-protoplasts or on hypocotyl-protoplasts cultured in 7.5 mM of IOA. IOA 2.5 mM inhibited colony formation of hypocotyl protoplasts and 5.0 mM IOA for leaf protoplasts. This research proved that IOA inhibited cell division of mungbean protoplasts var. Walet. <br />