To evaluate the resistance of hulless barley to barley yellow dwarf virus （BYDV）， 245 hulless barley genotypes were tested by inoculation with viruliferous aphids under field conditions in three calendar years. The results showed that the genotype 08-1280 was highly resistant to BYDV， and six genotypes （ZYM1289， Beiqing 6， ZDM4409， Ganqing 2， Zangqing 3000， ZYM1853） were resistant. 47， 173 and 18 genotypes showed moderately resistance， susceptibility and highly susceptibility to BYDV， respectively. The genotype 08-1280 was found carrying the resistance gene Yd2 based on genotyping using molecular marker. Moreover， 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 content between resistant genotype 08-1280 and susceptible genotype Kangqing 3 were investigated. At 10 days after inoculation， the contents of total phenol and proline were elevated in 08-1280， higher than that of Kangqing 3. At 30 days after inoculation， a higher increase of total phenol in resistant genotype， if compared to that of susceptible genotype， was detected， while an opposite result for the soluble sugar was observed； the soluble protein decreased dramatically in 08-1280， whereas increased in Kangqing 3. Collectively， this study provided elite germplasm resources and theoretical reference in breeding of resistant varieties and analysis of resistant mechanisms in hulless barley.