With the development of diverse agricultures worldwide, biofortified rice noted for its preferable marketability and palatability plays an important role in the world's agricultural economics and rice breeding programs. In this report, several M5 of T-DNA inserted lines derived from the donor cultivars, 'Hwayong' and 'Dongjin', were selected for high or low protein, high lipid and low amylose content, respectively. The coefficients and ranges of variation for the chemical constituents between M4 and M5 T-DNA inserted lines were evaluated in comparison with those of the donor varieties. Results indicated that T-DNA insertion might be an effective way to generate useful variations for chemical composition of rice grains which could be used for the development of biofortified rice cultivars.