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Conditions for high frequency embryogenesis from orange (Citrus sinensis Osb.) protoplasts
Authors:Shozo Kobayashi  Isami Ikeda  Hirofumi Uchimiya
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
Abstract:Using lsquoTrovitarsquo 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 (sim4×104/ml) and low mannitol concentrations (sim0.4 M). Two alternatives to achieve high frequency embryogenesis (sim70%) 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.
Keywords:Citrus sinensis Osb    nucellar callus  protoplast  somatic embryogenesis  inhibitor(s)  plant regeneration
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