كشت تعليقي پينه هاي رويان زا و نارويان زاي ليموآب

SUSPENSION CULTURE FROM EMBRYOGENIC AND NON-EMBERYOGENIC CALLI OF LIME (CITRUS AURANTIFOLIA S)

of were cultured. Non-embryogenic callus was produced with sweet lime (Citrus limettoides Tan) and Mexican lime (Citrus aurantifolia S) flaveado explants on MS I (Murashige and Skoog) medium containing I mg 1-1 2,4-D. (2,4-dichlorophenoxyacetic acid),0.1 mg 1-1 NAA (naphthaleneacetic acid) and 0.25 mg 1-1kinetin. Growth and cell division were not stimulate when the calli were subcultured on various media. In addition, non-ernbryogenic callus did not grow in suspension culture. Whereas, emberyogenic callus derived from unfertilized ovules of Mexican lime showed growth and cell division in MT (Murashige and Tucker) suspension medium supplemented with 1550 mg 1-1 glutamine and 500 mg 1-1 malt extract. Mean of counting in 3 replicated cultures at the beginning was 0.04 ± 0.002 * 10^4 and reached to 0.65 ± 0.03 * 10^4 after 21 d. In order to control internal bacterial contamination, flavedo explants were preculture or media treated by streptomycin and gentamycin. Antibiotics were not effective for disinfection, singly. Streptomycin (1000 ug 1-1)+ gentamycin (125 ug 1-1) mixture was the most effective treatment for disinfection in 60% of explants. Seventy percent of explants were clean when streptomycin (500 ug 1-1) + gentamycin (500 ug 1-1) mixture was used, but more than 50% of explants died due to antibiotic high concentration. Browning of explants increased with the presence of antibiotics in the media.