TRANSFORMATION OF GUS GENE INTO SOYBEAN CULTIVAR DT12 USING AGROBACTERIUM TUMEFACIENS

Abstract

The purpose of this study is to develop a culture system for Agrobacterium lumefaciens mediatedgenetic transformation in Vietnamese soybean, vector pPNT289 containing gus intron was used to transform into DT12 variety through multi-shoot induction method from maturely zygotic embryo tips. After sterilizing, mature seeds were soaked for 24 h, the embryonic tips were collected and cultured on MS + vitamin B5 + 3.5 mg/I BAP for 72 h in the dark. Induced embryonic tips were inoculated with Agrobacterium tumefadens strain EHA I 0 I containing the binary vector pPTN289 for 1.5, 2, 20 and 26 h. Our results showed that the best time for infection was 2 h with efficiency for the gus intron gene transformation, multi-shoot numbers reached up to 47% and the gus transformation efficiency reached up to 12.8%. Eight of regenerated lines: TOElT12-1, TOElT12-2, TOElT12-3, TOElTI2-4, TOElT12-5, TOElTI2-6, TOElTI2-7, TOElT12-8 were planted in the greenhouse at the Biologycal Experimental Station, Conhue, Tuliem, Hanoi. All of leaf, shoot and flower explants from eight TO lines showed the distinctive blue of gus gene. Total 330 seeds were harvested from eight TO lines. Analysis gus gen of leaf, shoot and flower explants from ten TI lines showed that 811 0, which contained gus gene, among 5 lines have gus gene in leaf, one line has gus in shoot and root and 2 lines have gus in leaf and flower.