Studies on breeding using protoplast-derived plants in rice (Oryza sativa L.), Koshihikari variety, 1: Fertile plant regeneration via rice protoplasts isolated from suspension cultures
1997
Mori, M. (Shiga-ken. Agricultural Experiment Station, Azuchi (Japan)) | Kitamura, H. | Uno, H. | Watanabe, K.
To improve Japonica rice (Oryza sativa L.) Koshihikari variety, by taking advantage of somaclonal variation, we attempted to obtain protoplast-derived Koshihikari plants using a suspension cultured system. The suspension cultures of Koshihikari were established from friable calli induced on N6 medium supplemented with 2 mg/l 2,4-D, 30 g/l sucrose and 3 g/l gelrite incubated for 60 days. Although the initiated calli of Koshihikari turned brown within 30 days of incubation, after another 20-30 days incubation without subculture under the same condition yellow localized friable calli were formed adjacent to the brown calli. The addition of glutamine to the R2 suspension medium was effective obtaining its suspension culture. Among 400 seeds plated on the medium, 4 cell lines were obtained. However, the efficiency of establishing cell lines was low. One cell line showing faster growth and higher plant regeneration was selected among the 4 cell lines and was used for protoplast culture. The addition of glycine and yeast extract to the regeneration medium was effective on the regeneration of Koshihikari. From 850 protoplast-derived colonies, 158 regeneration plants were obtained, 100 of which produced seeds. In the regeneration plants morphological variations were observed
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