Conditions for high frequency embryogenesis from orange (Citrus sinensis Osb.) protoplasts |
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Authors: | Shozo Kobayashi Isami Ikeda Hirofumi Uchimiya |
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Institution: | (1) Akitsu Branch, Fruit Tree Research Station, Akitsu, 729-24 Hiroshima, Japan;(2) Institute of Biological Sciences, University of Tsukuba, 305 Sakura-mura, Ibaraki, Japan |
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Abstract: | Using Trovita orange (Citrus sinensis Osb.) protoplasts isolated from 6-year-old nucellar callus, the effects of protoplast density and mannitol concentration on cell divisions and embryoid formation were examined.Somatic embryogenesis in nearly direct manner was observed only at a combination of low cell densities (4×104/ml) and low mannitol concentrations (0.4 M). Two alternatives to achieve high frequency embryogenesis (70%) were to either dilute the cells to lower densities, or to do serial transfers of cells to fresh medium.Orange protoplasts (cells) showed embryogenic potential, and repression of embryogenesis occurred when protoplasts were cultured at a high density and/or under high osmotic pressure. |
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Keywords: | Citrus sinensis Osb nucellar callus protoplast somatic embryogenesis inhibitor(s) plant regeneration |
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