Study on the fusion of sperm and somatic cells in loquat

The objective of this study was to fuse sperms of pollen and somatic cells of loquat, Eriobotrya japonica Lindl. The modified osmotic shock and density gradient centrifugation techniques were used to isolate loquat sperm nucleus. Meanwhile, protoplasts were separated from loquat leaves by enzymolysis. Then the sperms and the protoplast were fused. The results showed that the best fusion ratio was 5:1, and the fusion rate was 20.4%. Moreover, the optimum fusion activity occurred in a solution of 35% PEG (polyethylene glycol), Ca 2+of 60 mmol/L, and pH = 9. Under these conditions, the fusion rate was 22.9%. By the method of swallow liquid culture, we can obtain viable, dividing hybrid cells. We explored new methods of observing the fusion process. We also obtained hybrid cells which divided and grew into cell clusters.