Protoplast Cultures Of Oryza Sativa L. And Brachiaria Decumbens
| dc.contributor.author | DAUD, ZAINAH | |
| dc.date.accessioned | 2016-06-24T01:47:55Z | |
| dc.date.available | 2016-06-24T01:47:55Z | |
| dc.date.issued | 2015-09 | |
| dc.description.abstract | The tissue culture systems for the rice (Oryza sativa L.) cv. Fujisaka 5 and IRAT 13, and Brachiaria decumbens were established. Two stage surface sterilization techniques using 70 % ethanol and 20 % Clorox® was sufficient to obtain 83.3 % to 100 % aseptic seeds depending on species and rice cultivar used. A high seed germination percentage of 97.7 % was obtained for O. sativa cv. Fujisaka 5. While, O. sativa cv. IRAT 13 and B. decumbens showed low germination percentage of 28.0 % and 40.0 % respectively. Callus of O. sativa L. cv. Fujisaka 5 was initially induced from three explants, mature seeds, roots and leaves. The root explants and the mature seeds of O. sativa L. cv. Fujisaka 5 showed good response in callus production in term of biomass while no callus could be produced from the leaf explants MS solid medium + 2.0 mg/L 2,4-D was sufficient to induce formation of O. sativa callus. Embryogenic and non embryogenic calli were produced from the mature seed explants. However, the root derived callus of O. sativa L. cv. Fujisaka 5 produced fine root hairs on the callus surface after several subculture cycles. The calli produced were unorganized, yellowish white in colour with root formation and slow in growth. Somatic embryos were observed on MS medium containing either NAA or BAP alone or in combination, although the percentage of germinated embryos were low ranging from 0.1 – 0.8 % of total fresh weight. MS medium supplemented with 1.0 mg/L NAA was found to be effective for regeneration of O. sativa L. cv. Fujisaka 5. Somatic embryos of O. sativa L. cv. Fujisaka 5 regenerated into normal plantlets after subcultured into the PGR free MS medium. Cell suspension culture of O. sativa L. cv. Fujisaka 5 was established from two weeks old callus and showed the best result in liquid Amino acid medium + 2.0 mg/L 2,4-D + 0.2 mg/L kinetin (AA medium). The pipetting technique of subculturing was used to prepare a fine cell suspension cultures. The selection of the enzyme solution is very important for isolation of protoplast. Four concentration of enzyme solution containing Cellulase Onozuka R10 and Macerase Pectinase were tested. Protoplasts were isolated from cell suspension culture of O. sativa cv. Fujisaka 5 and IRAT 13, while leave mesophyll and callus were used for B. decumbens using 1.0 % (w/v) Cellulase Onozuka R10, 0.1 % (w/v) Macerase pectinase, 0.1 % (w/v) Polyvinylpyrrolidine-10, and 5 mN MES dissolved in CPW 13M at pH 5.8. The overnight incubation method was used and enable to produce sufficient cells for protoplast culture. However, protoplast culture using liquid culture method and agarose bead techniques failed to form microcolonies | en_US |
| dc.identifier.uri | http://hdl.handle.net/123456789/2180 | |
| dc.subject | Protoplast Cultures Of Oryza Sativa L. | en_US |
| dc.subject | And Brachiaria Decumbens | en_US |
| dc.title | Protoplast Cultures Of Oryza Sativa L. And Brachiaria Decumbens | en_US |
| dc.type | Thesis | en_US |
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