Development of regeneration system for biolistic-mediated transformation for abaca
2007
Aspuria, E.T. | Garcia, L.Z. | La'o, L.A. | de Leus, G.R., Philippines Univ. Los Banos, College, Laguna (Philippines). Crop Science Cluster
Explants of the four abaca cultivars previously inoculated onto MC + 5 mg/L BA were introduced to form meristematic buds (scalps) on MS + 1 microM IAA supplemented with either 50 MicroM BA or 1 MicroM TDZ. A comparative study on the four abaca cultivars indicated that 'Parang', 'Inoza' and 'Tangongon' indicating that TDZ is a more effective cytokinen. However, relatively similar rate of shoot formation was observed in TDZ and BA containing medium. Globule formation was observed 4-5 weeks after inoculation in modified 1/2 MS +5 micro M 2,4-D+ 1 micro zeatin. 'Abuab' showed the earliest to form and release globules from scalps. Fifty to sixty globules from each of the 4 abaca cultivars were collected and maintained fro 3 weeks in fresh culture medium (globule formation) for the establishment of embryonic cell suspension cultures. 'Inosa' produced the highest number of cells but with low % cell viability. Mature cells of the 4 abaca cultivars were induced to undergo somatic embryogenesis in induction media consisted of MS + BA further supplemented with ABA alone or in combination with malt extract. However, no further growth was observed after 3-4 weeks of incubation. On the other hand, globules inoculated in 3, 4, 5 and 7 days, the highest number of cells released was observed at 5 days after incubation except for 'Parang' cultivar which attained its peak after four days of incubation. Cell viability ranging from 59 to 87.1% was observed at 4 to 5 days of incubation in all abaca cultivars. On MS = 9.1 micro M zeatin medium, cells of 'Tangonon' and 'Abuab' cultivars developed into somatic embryos even without passing through PGR-free medium. However, fully developed somatic embryos obtained from 'Tangonon' cells continued to proliferate but had not yet germinated at present. In 'Abuab' cultivar, only root formation was observed indicating the absence of embryo bipolarity.
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