Acquirement of CRY8DB Transgenic Tall Fescue (Festuca arundinacea Schreb.) by Agrobacterium tumefaciens to Develop Resistance Against Pentodon idiota Herbest.

Abstract

Tall fescue plants are widely exposed to white grubs. Regarding the rate of damage caused by the white grubs to tall fescue and difficulty of its ecological and economical control, production of resistant cultivars is a priority. In this experiment, for the first time, we report production of transgenic lines resistant to white grub using CR8DB gene. For this, mature seeds were placed on MS basal medium with 0-15mgL-1 2,4-D for callogenesis and 0-1.75mgL-1 BA for regeneration. 'Asterix' (54.11%) in 7.5 and 'Talladega' (52.53%) in 10mgL-1 2,4-D showed maximum callogenesis. Regeneration percentage was higher in 0.5mgL-1 BA. Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pCAMBIA 1301 with CRY8DB gene, which contains HPTII gene and uidA and various types MS media were used for transformation of calli. The highest percentage of gus enzyme activity and hygromycin resistance in calli was related to the modified medium type 11. The PCR and RT-PCR analysis was done to confirm the presence and expression of the target gene in transgenic 5 lines in 'Asterix' and 3 lines in 'Talladega'. According to bioassay, larvae mortality of 91.66% was observed in transgenic plants, whereas it was 15.52% in control plants.