Studies on factors involved in transfer of foreign gene by pipetting Agrobacterium into spikelets of wheat

Abstract

Studies were conducted on wheat cv. UP 2338 for various factors involved in gene transfer by pipetting Agrobacterium suspension into florets using indirect pollen system. Pollen grains showed germination after 20 min of pollination that was considered as an optimum time for Agrobacterium inoculation. After 20 min of pollination 5111 of Agrobacterium (LBA4404 containing binary vector pB1121) suspension was pipetted into the florets. Gram negative staining for Agrobacterium showed presence of pink colour after 20 min of Agrobacterium inoculation and at later stages some parts of pistil and pollen took pink colour. The seeds obtained after Agrobacterium inoculation into the florets showed 5.5% putative transformation frequency when screened against 500 mgll kanamycin. Based on phenotypic observations it can be concluded that it is possible to transfer foreign gene via pollen through Agrobacterium.