Studies on Culture of Cells and Tissues of Crop Plants : I. Survey on enzymatic isolation and culture of rice leaf sheath protoplasts

Japanese Journal of Crop Science

Masako OHSHIMA¹  Susumu TOYAMA¹ 

关键词: BSA;    イネの葉鞘;    Cell activity;    MS培養液;    Cellulase "Onozuka" R-10;    細胞活性;    Protoplasts;    セルラーゼ"オノヅカ" R-10;    MS-medium;    Rice leaf sheath;    プロトプラスト;   

DOI  :  10.1626/jcs.58.103

学科分类：农业科学（综合）

来源: Nihon Sakumotsu Gakkai / Crop Science Society of Japan

【 摘 要 】

To establish a method for isolation and culture of rice protoplasts, enzyme solution compositions, culture conditions, and the selection of cultivars and tissues as materials were examined. The results suggested that the 1st and 2nd leaf sheath of 6-day old seeldings, which were cultured in the medium containing only minerals and vitamines of MS medium under 23°C, 16.2 W/m2, and light length 14 h, were found suitable as materials. The materials were treated without shaking for 4 h in an enzyme solution (pH 5.6) containing 0.25% macerozyme R-10, 1% cellulase "Onozuka" R-10 and 1% BSA in a MS medium which contained MS-minerals, MS-vitamines, 1.0 mg/l 2, 4-D, 2.5 mg/l kinetin and 10 g/l sucrose. The isolated leaf sheath protoplasts were cultured in a MS medium (0.42 M) containing 1% BSA. All protoplasts from 11 cultivars kept their cell activity through the culture for 2 weeks or more. The addition of BSA in both the enzyme solution and culture medium was especially effective for the maintenance of protoplast viability. The vacuoles developed in the protoplasts during the culture period; after 5 days in the culture the protoplasts enlarged to a diameter 1.75 times greater compared with that just after the isolation. The chloroplasts in protoplasts began to dedifferentiate during the culture, and both chloroplast size and chlorophyll content decreased. Protoplast division was not observed in this medium throughout the culture process.

【 授权许可】

Unknown