To evaluate the resistance of hulless barley to barley yellow dwarf virus (BYDV) , 245 hulless barley materials were tested three?times by inoculation of viruliferous aphids under field conditions. The results showed 08-1280 possessed highly resistance to BYDV, six materials（ZYM1289, Beiqing 6, ZDM4409, Ganqing 2, Zangqing 3000, ZYM1853）were resistant. 47,173, and 18 materials showed moderately resistance, susceptibility and highly susceptibility to BYDV, respectively. Molecular marker detection indicated that 08-1280 carried Yd2 resistant gene.To gain a better understanding of physiological mechanism of hulless barley against BYDV infection, the differences in total phenol, proline, soluble sugar and soluble protein changes between resistant material ‘08-1280’ and susceptible material ‘Kangqing 3’ were investigated. We found that the physiological response to BYDV varied with resistance and time after inoculation in hulless barley. 10 days after inoculation, the total phenol and proline of high resistant material showed increases significantly higher than high susceptible one. 30 days after inoculation, the increase of total phenol of high resistant material were significantly higher and the increase of soluble sugar was significantly lower as compared with high susceptible material. Compared with the control, soluble protein decreased dramatically in high resistant material, but increased in high susceptible one at 30 days after inoculation. This study provided excellent resources and theoretical reference for breeding?resistant?varieties and clarifying?resistant?mechanisms in hulless barley. This study provided excellent resources and theoretical reference for breeding?resistant?varieties and clarifying?resistant?mechanisms in hulless barley.