Clean DNA transformation is the technology of introducing exotic gene expression cassette into plant genome via particle bombardment, which can eliminate the disadvantageous impact of vector backbone sequence on transgenic plant fundamentally. Gene 2mG2epsps is an important glyphosate herbicide resistance gene with important breeding value. The effect of glyphosate on rice callus growth and differentiation was studied using japonica rice Nipponbare as material, and 2mG2epsps gene cassette was transformed into rice by clean DNA transformation. Results showed: 1) The growth and differentiation of rice callus can be notably inhibited by glyphosate. The regeneration frequency of green plantlet decreased significantly to 18.97%, comparing with that of the control 71.67%, at 2 mmol/L glyphosate; 2) During rice transformation with 2mG2epsps gene expression cassette via particle bombardment, removal of the screening agent from regeneration medium is beneficial to the differentiation of the glyphosateresistant calli, which were screened out under glyphosate selection with transformation frequency at 17.20%. Southern blot analysis revealed that the 2mG2epsps gene cassette was all integrated into rice genome in single copy and 52.17%（12/23)transgenic lines can render 12-50 mmol/L glyphosate. The present research provided the foundation for breeding appliance of the glyphosate resistant transgenic rices.
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