*Central Potato Research Station, Muthorai-643 004, Udhagamandalam, The Nilgiris, Tamil Nadu
Department of Biotechnolgy, School of Biotechnology and Genetic Engineering, Bharathiar University, Coimbatore-641 046, Tamil Nadu
Online published on 17 June, 2017.
In the present study, optimization of in planta genetic transformation of Solanum melongena L. var. CVK (Covai Vari Kathari) was attempted using seed as an explant and Agrobacterium tumefaciens strain LBA4404 containing the gene construct 35S: nptII: RCI2A in the binary vector pBI121. The Minimum Inhibitory Concentration (MIC) of kanamycin on S. melongena var. CVK seed germination was found to be 200 mg/L. Among the different seed pretreatment experiments, soaking + scarification + preculture (24 h) produced the best transformation efficiency of 5.24%. Co-cultivation media amended with 200 μM of acetosyringone gave a transformation efficiency of 5.2% and a sonication time of 15 minutes gave the best transformation efficiency of 6.19%. Among the different methods of in planta transformation employed, immersion in an Agrobacterium suspension+ vacuum infiltration gave a maximum transformation efficiency of 7.62%. Molecular confirmation of transformation was done using primers for kanamycin resistance gene (nptII) and transformation efficiencies were calculated based on the mean number of PCR positive seedlings.
Acetosyringone, co-cultivation, minimum inhibitory concentration, sonication, transformation efficiency, vacuum infiltration
