A simple method for isolation of haploid protoplasts from forms of rice multishoots
DOI:
https://doi.org/10.7124/bc.000205Abstract
One-month old sterile plantlets grown on the 0.3 M sucrose medium were used for protoplast isolation. The young part of leaves were cut onto small pices (1 mm) and left for over night incubation in the 2 % v/v driselase solution containing glucose 0.4 M. Isolated protoplasts were purified by two-phase centrifugation (the upper phase contains solution of salts, the lower one — 0.4 M of sucrose and other nutritional constituents). The purified protoplasts were suspended between two phases.References
Chu CC, Wang CC, Sun CS et al. Establishment of an efficient medium for anther culture of rice through comparative experiments on nitrogen sources. Sci Sinica. 1975; N 5:659-668.
King P, Potrykus I, Thomas E. In vitro genetics of cereals: problem and perspectives. Physiol Veg. 1978; 16(2):381-399.
Dunstan DI, Short KC, Dhaliwal H, Thomas E. Further studies on plantlet production from cultured tissues ofSorghum bicolor. Protoplasma. 1979; 101(4):355-361.
Thomas E, King P, Potrykus I. Improvement of crop plants via single cells in vitro an assessment. Z. Pflanzenzucht. 1979; 82(1):1-30.
Vasil I. K. Regeneration of plants from single cells of cereals and grasses. Genetic engineering in eukaryotes. Eds P. F. Lurquin, A. Kleinhofs. New York : Plenum PubL Co., 1983:233-252.
Gleba YuYu, Sitnik KM. Cell engineering of plants. Kiev, Naukova Dumka, 1984; 160 p.
Murashige T, Skoog F. A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum. 1962. 15(3):473-497.