Plant regeneration from protoplast-derived callus of rice (Oryza sativa L.)

Abstract

Protoplasts isolated from cultured rice cells of an A-58 cytoplasmic male sterile line (A-58 MS)(Oryza sativa L.) were used to investigate the regeneration of rice plants. A cultured cell line (T3) of A-58 MS with a high growth rate and dense cytoplasm was selected. About 10% of the protoplasts prepared from this established cell line plated in RY-2 (a new medium) formed colonies. The calli formed shoots and roots in the regeneration medium and developed into whole plants.Protoplasts also were prepared from suspension cultures of 25 other varieties of rice using the same methods. The protoplasts isolated from two of the 25 varieties, Fujiminori and Toyotama, had high rates of cell division in RY-2 medium. Only protoplastderived calli from Fujiminori, produced whole plants in the regeneration medium.

Regeneration of variant plants from rice (Oryza sativa L.) protoplasts derived from long term cultures.

“…In previous reports, suspension callus was used as the donor material (Fujimura et al 1985;Yamada et al 1986;Toriyama et al 1985;Abdullah et al 1986;Kyozuka et al 1987). In this experiment, the callus sub-cultured on a solid medium was useful as material for protoplast isolation.…”

“…Recently, successful regeneration of whole plants from rice protoplasts has been reported (Fujimura et al 1985;Oono 1986;Yamada et al 1986;Coulibaly and Demarly 1986;Toriyama et al 1986;Abdullah et al 1986;Kyozuka et al 1987). However, the frequency of plant regeneration was often low.…”

Proteome and phosphoproteome analysis of chromatin associated proteins in rice (Oryza sativa)

“…To obtain high quality chromatin, rice suspension culture cells [24,25] were used for chromatin isolation and purification in our studies. The protoplasts were released from suspension cells by enzymatic digestion of the cell wall as reported [26] for 8 h.…”

“…For protoplast isolation, cells were harvested 4 days after subculture. The protoplasts were generated using a method as reported by Yamada et al [26] with modifications. In brief, suspension cultured cells were added to filter-sterilized enzyme solution containing 2.5% Cellulose RS (Onozuka RS), 1% Macerozyme R10 (Research Products International), 0.4 M mannitol, 80 mM CaCl 2 , 0.125 mM MgCl 2 , 0.5 mM MES, and B5 organic medium plus 2.0 mg/L 2,4-D (pH 5.6).…”

Callus induction and plant regeneration via leaf segments of three accessions of African rice (Oryza glaberrimaStued.)

“…The ability to regenerate plants from callus is influenced by physiological factors as well as the genotype of the plant (Henry et al, 1994). The regeneration of plants of some cereal crops such as bread wheat (Redwayet al, 1990;Vasilet al, 1990), barley (Luhrs and Lorz, 1987),rice (Yamada et al, 1986) and maize (Duncan et al, 1985), from callus have been documented. In rice, there are reports on successful plant regeneration from explants such as coleoptile (Oinam and Kothari, 1995), root tips (Sticklen, 1991) …”