HomeInternational Rice Research Notesvol. 22 no. 2 (1997)

High-Frequency Plant Generation in Wild Species of Oryza

G S. Oinam | S L. Kothari

Discipline: Agriculture



We are developing a system for Plant regeneration from wild species of Oryza. In recent work, dehusked seeds were surface-sterilized with 0.1% (w/ v) HgCl2 solution for 5 min and rinsed four times in sterile distilled water. Sterilized seeds were kept for germination on MS medium devoid of growth hormones in light (1600 lux) at 25ºC. The endosperm and radicle were excised from 7-d-old seedlings, which were then inoculated on MS medium supplemented with 6.0 mg BAP L-1. Multiple shoots or micropropagules developed at the base of shoots were used for callus initiation. Segments (0.5-1.0 cm long) cut from their white tissue were inoculated on LS medium containing 2.5 mg 2,4-DL-1, 3%) sucrose, and pH was adjusted to 5.8. The cultures were kept in the light. Embryogenic calli from the primary cultures were subcultured on the same medium and then transferred to regeneration medium based on MS salts supplemented with 2.0 mg BAP L-1 and 0.5 mg NAAL-1. All media were solidified with phytagel (0.2% Sigma) and growth hormones avoided. Rooted plantlets were transferred to pots.