Abstract

Presently, genetic modification of wheat is carried out through gene gun andAgrobacterium mediated transformation that involves callus phase and is restricted to the tissue culture responses of genotypes. Therefore, a protocol for genetic transformation of wheat without involving tissue culture procedures was established. Imbibed seeds of wheat were sterilized, incised through apical meristem, wounded and inoculated with Agrobacterium tumefaciens containingGUS and NPT-Il genes in its plasmid. Treated seeds were co-cultivated for three days followed by washing and placement of seeds in Petri dishes for regeneration. The regenerating seeds were subjected to kanamycin selection for a period of 4 weeks. Two hundred and eight kanamycin resistant plants were produced out of 1850 inoculated seeds. Immature seeds of kanamycin resistant plants were assessed for GUS expression and 27% of the total plants were found with positiveGUS activity. Polymerase chain reaction (PCR) amplification of NPT-II gene using specific primers confirmed integration of transgene in 26% of the kanamycin resistant plants. The developed protocol is cheap, efficient and genotype independent without involvement of any tissue culture procedure.

Key words: Wheat, in planta transformation, apical meristem, GUS.

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