Study on Efficient Protoplast Isolation System for Dahlia pinnata Petals
In order to solve the problems of slow conventional breeding process,imperfect breeding system of modern biotechnology,and immature system of genetic transformation and gene function verification in Dahlia pinnata,Dahlia pinna-ta petals were used as experimental materials,and the effects of material treatment,mannitol concentration,enzymolysis time,enzymolysis combination and purification method on the yield and viability of Dahlia pinnata protoplasts were studied.The results showed that the most effective method for isolating Dahlia pinnata petal protoplasts involved cutting the petal tissue into 1 mm strips using the filament-cutting technique.The strips were then placed in an enzyme solution consisting of 1.00%cellulase,0.50%macerozyme,0.40%pectinase,and 1.00 M mannitol for 10 hours.The highest yield of dahlia petal protoplasts was 5.46×106/mL,and the highest viability was 88.83%.An efficient,stable and rapid system for isolating dahlia petal protoplasts has been established,in order to provide a good experimental platform and technical support for subsequent analysis of Dahlia pinnata somatic cell fusion and gene function.
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