Volume IX No.2 .TRANSTORMASI GTINETIK LIMA VARIETAS KEDELAT

Article (PDF Available) · August 2004with30 Reads
Abstract
ABSTRACT AGROBACTERIUM-MEDIATED TRANSFORMATION OF FIVE SOYBEAN CULTIVARS. The objective of this study was to evaluate the efficiency of Agrobacterium-mediated tranformation using explants of five cultivars of soybeans. The study was conducted in the Plant Transformation Core Research Facility, Center of Biotechnology, University of Nebraska, Lincoln, Nebraska, USA from July – December 2003. Experiments were arranged in randomized block designs with 5-10 replications, 4 explants within each experimental unit. Three strains of Agrobacterium tumefaciens were used, i.e., C58C1, EHA101, and NTL4-Cry5. Explants were cotyledonary nodes of cultivars Jaya Wijaya, Krakatau, Slamet, Tampomas, and Wilis. Cotyledonary-node explants, prepared from germinated seeds, were inoculated with Agrobacterium, incubated in co-cultivation media for 3 days, washed, cultured in media of shoot-initiation, shoot-elongation, and rooting for 4, 4-6, and 2 weeks respectively. The media of shoot initiation and elongation contained herbicide glufosinate as the selecting agent. At 31 days after inoculation, shoots were produced from explants of all five cultivars evaluated. The proportion of explants producing shoots ranged from 31% (cultivar Jaya Wijaya) to 76% (cultivar Slamet). Transgenic soybeans were produced from all five cultivars evaluated with efficiency ranged from 2,6% to 6,5%. Transgenic soybeans were only produced from explants inoculated with EHA101 and NTL4-Cry5, not with C58C1. Key words : Agrobacterium, Glycine max, genetic engineering