Transformation and regeneration protocols for Brassica napus varieties adapted to agronomic conditions prevailing in India were standardized. Five different varieties of B. napus (BO-15, GSL-1, HNS-8, H-18, ISN706) and a hybrid of ISN706 and GSL-1 were tested for regeneration response. B. napus var GSL-1 showed a very high frequency of regeneration (98%). Regeneration response of some of the varieties was improved by alterations in the composition of media. Efficient transformation protocol was developed for the variety GSL-1 with a binary vector containing nptll and bar genes conferring resistance to kanamycin and herbicide phosphinothricin as selection marker and a gus-Int gene as a reporter. A new construct that contained Ds border, bar gene as integration marker and a mutant csr-1 gene conferring resistance to chlorsulfuron as an excision marker was developed for transposon-mediated gene tagging. This Ds-construct was introduced into B. napus genetic stock (ISN706 GSL-1 Rft/-, Rfp/-) harboring fertility restorer (Rf) genes for two different CMS systems namely ‘tour’ (Rft) and ‘polima’ (Rfp).