Protoplast Fusion of Cosmos bipinnatus and Cosmos sulphureus

摘要

[Objective] The aim was to study appropriate fusion conditions of protoplasts from 2 species Cosmos bipinnatus and Cosmos sulphureus. [Method] The protoplasts were separated from the blossom petals of Cosmos bipinnatus and Cosmos sulphureus availed with the function of pectinase and cellulase,and the fusion of two protoplasts was performed with PEG. [Results] A great deal of free protoplasts were obtained from Cosmos bipinnatus by using 6% mannitol + 1.3% cellulase + 1.2% pectinase,and from Cosmos sulphureus by using 10% mannitol + 1.3% cellulase + 1.2% pectinase. Five hours after of enzymolysis treatment,large amount of dissociated protoplasts were observed in the solution. Protoplast fusion was be obtained by using 30% PEG fusion liquid and 0.1mol /l CaCl2 solution (pH value =9.5) . [Conclusion] Our study established an optimal separation and fusion system of protoplasts from different coreopsis species,laying foundation for the breeding of novel coreopsis varieties.