Biolistic transformation ofcry 1Ac gene in eggplant (Solanum melongenaL.)

M. K. Sidhu, A. S. Dhatt, J. S. Sandhu, S. S. Gosal
2014 International Journal of Agriculture Environment and Biotechnology  
Among different explants, cotyledon showed highest regeneration and number of buds per explant on all media combinations. However, highest regeneration potential was observed on MS fortified with 3.0 mgl -1 BAP (80.36%), but number of buds per explant were highest (21.76) on MS media supplemented with 2.5 mgl -1 BAP+ 1.0 mgl -1 Kin. Plant regeneration of cotyledons was completely checked with a selection pressure of 15ppm to 25ppm hygromycin for untranformed explants. But the selection efficacy
more » ... selection efficacy of transformed tissue was 19.56% with 20ppm and 66.66% with 15 ppm hygromycin. GUS gene was expressed in 76.92% bombarded explants with 2 to 7 GUS spots per explant confirming the presence of transformed tissue. GUS gene was also visualized in 66.66% regenerated shoots from bombarded tissue. PCR analysis of putative transgenic eggplant plants with the specific primers confirmed the presence of Cry1Ac gene with 1.2 KB product size (falling between 1KB and 2KB bands of 10KB DNA ladder) in two of the four plants. Highlights • Cotyledon of genotype BL-5 had highest regeneration potential and number of buds per explant on MS media supplemented with 2.5 mgl -1 BAP+ 1.0 mgl -1 Kin. • Plant regeneration of cotyledons was completely checked with a selection pressure of 15ppm to 25ppm hygromycin for untranformed explants. • The selection efficacy of transformed tissue was 19.56% with 20ppm and 66.66% with 15 ppm hygromycin. • GUS gene was expressed in bombarded explants as well as in regenerated putative transgenic explants. • PCR analysis of putative transgenic eggplant plants with the specific primers confirmed the presence of Cry1Ac gene.
doi:10.5958/2230-732x.2014.01375.8 fatcat:fbu7jggu5zbfjho63st6t5ck4i