A Novel Method of Plant Regeneration from Suspension Culture Protoplasts of Sugarcane

Abstract

Summary Protoplasts were released from cell suspension cultures of two genotypes (87-588 and F177) of sugarcane ( Saccharum spp. hybrids). They were embedded in an agarose block with PRM medium, over which 5 mL nurse cell culture was flooded. The nurse cell culture was changed at weekly intervals to reduce osmotic pressure and to readjust the cell density. Genotype 87-588 micro-colonies formed on the agarose block within 1 month. After having grown to a larger size, they were transferred to a solid MS based medium where they took on a rectangular shape (1.2x2.8cm). This proto-callus (from 87-588) was surrounded by a 0.8-cm thick wall composed ofF 177 callus covered with newly differentiated green shoots. By using the nurse green shoot's differentiating factor carry-over idea, the enclosed 87-588 callus was induced to undergo meristematic growth. After 30 days, the treated-callus was placed on a differentiation medium where many green buds were regenerated within a 1-month incubation. Using a similar method, plant regeneration from protoplasts of F177 was also successful.