Objectives: To verify the reliability of screening positive seedlings of transgenic rice by using the agar solid medium with hygromycin and determine the optimal concentration of hygromycin.Methods: A sort of transgenic plant with hpt and GUS selection marker gene drived by 35S promoter was chosen as experimental material in the study.The transgenic seeds were sowed after sterilized on the 0.5%agar solid medium with a concentration gradient hygromycin of 0、25、50、75、100 mg/L and the growth situation of seedlings were observed,especially roots condition.Meanwhile,PCR verification of hpt gene was operated after extracting the total DNA,GUS histochemical staining was analyzed with the white roots,and the hygromycin resistance detection of detached leaves was also carried out by dipping in the solution with 100 mg/L hygromycin.The wild type plant was regarded as the experiment control.Results: The transgenic seedlings grown on the hygromycin solid medium with normal root system could obtain the accurate electrophoretic band in PCR verification of hpt gene,roots could been stained by X-Gluc,leaves could also remain viridian,but wide type and negative seedlings couldnt.Eventually,the study showed that using the agar solid medium with 25 mg/L hygromycin was enough to screen the positive seedlings of transgenic rice rapidly.Conclusions:The study verified and provided a rapid handy economical method to screen the positive transgenic seedlings with hygromycin resistance.It was useful for sifting transgenic plants in researching the function of genes by genetic transformation.