LIPOSOME-MEDIATED DELIVERY OF CALCEIN INTO CUCUMBER SUSPENTION PROTOPLASTS
Tang Hui-xian Jia Shi-rong (Biotechnology Research Center,Chinese Academy of Agricultural Sciences) Yen Chang-cheng Zhao Nan-ming (Department of Biological Science and Biotechnology,Tsinghua University)
Liposome-encapsulated calcein prepared by 'dehydration-rehydration method' was successfully introduced into cucumber suspen-sion protoplasts by means of PEG-induced fusion or liposome/protoplast co-cultivation.By PEG treatment,the percentage of cells with liposome-mediated delivery of calcein was around 80-90%.among which 50-60% cells showed strong,even distributing fluore-scence.In the case of liposome/protoplast co-incubation (half an hour),the percentage
of fluorescent cells reached more than 95%.However,the fluorescence was relatively weak and dot-distributed within cells.The liposo-me was broken after 3-4 days in culture so that the dotted fluorescence became evenly distributed.Cucumber protoplasts uptaking liposomes successively underwent sustained cell division,formed callus and embryoids which eventually regenerated into shoots and roots.
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