The research object of this study is “ML7113” wheat leaf, which is used to isolate protoplast with enzyme hydrolysis method. Three main effectors—the concentration of mannitol, enzymolysis time and centrifugal forc...The research object of this study is “ML7113” wheat leaf, which is used to isolate protoplast with enzyme hydrolysis method. Three main effectors—the concentration of mannitol, enzymolysis time and centrifugal force, affect the production and vitality of protoplast. While the production and vitality of wheat protoplasts were detected by the hemacytometer and the FDA staining respectively. Results showed that, with the increasing concentrations of mannitol during 0.2 M - 0.4 M, protoplast yield increases and when the concentration is 0.4 M, the protoplast vitality can be up to 95%;with the extension of enzymolysis time in 2 h to 8 h, protoplast yield reaches a maximum in 6 h, but its vitality achieves the maximum in 4 h;considering a combination of these two factors impacting on protoplast, we obtain the best time to digest for 4 h;meanwhile, with the increasing of the centrifugal force from 500 rpm - 2000 rpm, its comprehensive effect of protoplast vitality and yield is the highest when the centrifugal force is 1000 rpm for 2 min (replicated three times). So 0.4 M mannitol, 4 h enzymolysis time and 1000 rpm for 2 min centrifugal force are the best separation condition.展开更多
The identification,separation and analysis of individual living cells can be used to analyze the heterogeneity and operation mechanism of living systems.The study of fruit development is based on the extraction of act...The identification,separation and analysis of individual living cells can be used to analyze the heterogeneity and operation mechanism of living systems.The study of fruit development is based on the extraction of active single cells.In this study,we investigated the effects of different enzymes and enzymatic hydrolysis times on the extraction of single cells from the‘Fuji’apple(Malus×domestica Borkh.‘Fuji’).The results showed that the extraction of single cells in apple flesh was a suitable method when 0.1%macerozyme was used and the enzymolysis time was 0.5 h.Fluorescent brightening agent VBL staining showed that the cell wall was intact,while fluorescein diacetate FDA and azo dye Evans blue staining indicated that the extracted single cells were active.The extracted single cells could be further used as materials for protoplast extraction.展开更多
文摘The research object of this study is “ML7113” wheat leaf, which is used to isolate protoplast with enzyme hydrolysis method. Three main effectors—the concentration of mannitol, enzymolysis time and centrifugal force, affect the production and vitality of protoplast. While the production and vitality of wheat protoplasts were detected by the hemacytometer and the FDA staining respectively. Results showed that, with the increasing concentrations of mannitol during 0.2 M - 0.4 M, protoplast yield increases and when the concentration is 0.4 M, the protoplast vitality can be up to 95%;with the extension of enzymolysis time in 2 h to 8 h, protoplast yield reaches a maximum in 6 h, but its vitality achieves the maximum in 4 h;considering a combination of these two factors impacting on protoplast, we obtain the best time to digest for 4 h;meanwhile, with the increasing of the centrifugal force from 500 rpm - 2000 rpm, its comprehensive effect of protoplast vitality and yield is the highest when the centrifugal force is 1000 rpm for 2 min (replicated three times). So 0.4 M mannitol, 4 h enzymolysis time and 1000 rpm for 2 min centrifugal force are the best separation condition.
基金supported by The National Key Research and Development Program of China (2016YFD0201120)Shandong Province Fruit System Job Expert (SDAIT-06-05)
文摘The identification,separation and analysis of individual living cells can be used to analyze the heterogeneity and operation mechanism of living systems.The study of fruit development is based on the extraction of active single cells.In this study,we investigated the effects of different enzymes and enzymatic hydrolysis times on the extraction of single cells from the‘Fuji’apple(Malus×domestica Borkh.‘Fuji’).The results showed that the extraction of single cells in apple flesh was a suitable method when 0.1%macerozyme was used and the enzymolysis time was 0.5 h.Fluorescent brightening agent VBL staining showed that the cell wall was intact,while fluorescein diacetate FDA and azo dye Evans blue staining indicated that the extracted single cells were active.The extracted single cells could be further used as materials for protoplast extraction.
