The purpose of this study is to measure of growth and chlorophyl in barley exposed by X-ray. Barley seed was soaked 24h duration in water, then was classified into two group; pre-seed germination group (Pre-G) or post-seed germination group (Post-G). Also, divided as control subgroup and experimental subgroup(10Gy, 20Gy, 30Gy) in each group. Experimental subgroups were exposed by X-ray using linear accelerator (Clinac IS, VERIAN, USA). Expose condition was 6 MV X-ray, SSD 100 cm, 18×10 cm, 600 MU/min. Length was measured every day for 10 days and 10th day for weight. Chlorophyl was analyzed using spectrophotometer(uv-1800, shimadzu, japan) in l0th day. Data analysis was performed using SPSS ver 22.0(Chicago, IL, USA), ANOVA test (Dunnett_T3) between control subgroup and experimental subgroup in group and Independent T-test between Pre-G and Post-G in subgroup. In Pre-G, length of barley was significantly difference between control and 30Gy in 4th day (4.3 vs. 1.5, p= 0.011). Length of 30Gy was statistical difference with control(10th day; 14.4 vs. 6.3, p < 0.01), and was not in 10Gy or 20Gy in all day. In experimental subgroup, length was shorter as increasing radiation dose. In Post-G, length of barley was not difference statistically between control and experimental subgroup in first day, but more difference between two subgroup with increasing duration after exposing. Length of experimental subgroup was shorter significantly compared with control in 10th day, and no significant difference between experimental subgroup. Density of chlorophyl was increasing with increasing radiation dose in Pre-G and Post-G. Chlorophyl density of control was lower than 30Gy; 0.26ppm in Pre-G, 0.29ppm in Post-G). Growth and chlorophyl of barley was effected by X-ray. It is expected to be used as basic data for future radiobiological research.