High frequency plant regeneration from rice protoplasts by novel nurse culture methods |
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Authors: | Junko Kyozuka Yasuyuki Hayashi and Ko Shimamoto |
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Institution: | (1) Plantech Research Institute, 1000 Kamoshida, Midori-Ku, 227 Yokohama, Japan |
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Abstract: | Summary Novel nurse culture methods have been developed for plant regeneration from protoplasts of rice (Oryza sativa). The nurse culture methods use the agarose-bead type culture in combination with actively growing nurse cells that are either in the liquid part of the culture or inside a culture plate insert placed in the centre of the dish. Protoplasts isolated from either primary seed calluses or suspension cultures of various callus origins, divided and formed colonies with a frequency of up to 10% depending on the protoplast source and the genotype. The presence of nurse cells was absolutely required for the induction of protoplast division. Plants were regenerated from protoplast-derived calluses of five tested cultivars with a frequency of 17%–50%. Close examination of the plant regeneration process suggested that plants are regenerated through somatic embryogenesis from protoplast-derived calluses. Over 300 protoplast-derived plants were transferred to either pots or the field and are being examined for karyotypic stability and various plant phenotypes. |
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Keywords: | Gramineae Oryza sativa Protoplast Nurse culture Somatic embryogenesis |
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