Abstract: In order to study the cryopreservation effect of dilutions containing different sugars on Black-bone sheep spermatozoa, 12 healthy adult Black-bone rams were selected for sperm collection.Semen were collected from 5 to 7 rams every time.Sperm cryopreservation was performed with Tris-citric acid-yolk diluent containing fructose（D1 diluent group）, Tris-citric acid-yolk diluent containing glucose（D2 diluent group） and lactose-yolk diluent containing lactose（D3 diluent group） by two-step dilution method. The sperm motility, viability, plasma membrane and acrosome integrity indexes before frozen and after thawing were measured, respectively.The percentage of motile sperm, viable sperm, sperm with intact plasma membrane and sperm with intact acrosome were calculated. The differences of the above indexes between fresh semen and thawed semen, as well as thawed semen containing three diluents were analyzed and compared. The results showed that the percentage of motile sperm, viable sperm, plasma membrane intact sperm and acrosome intact sperm in fresh semen were（89.23±4.54）%,（64.65±6.78）%,（72.59±9.07）% and（82.79±7.87）%, respectively.After diluting, cooling, freezing and thawing, compared with fresh semen, the percentage of motile sperm, viable sperm, plasma membrane intact sperm and acrosome intact sperm extremely significant decreased by 80.46%, 67.21%, 60.35% and 39.73%, respectively（P＜0.01）.The percentage of motile sperm after thawing in D1 diluent group（33.64±4.37%） was significantly higher than that in D2 and D3 diluent groups（9.26±2.42%, 9.42±4.63%, P＜0.05）, but there was no significant difference in the percentage of motile sperm between D2 and D3 diluent groups（P＞0.05）.The percentage of viable spermatozoa after thawing was highest in the D1 diluent group（33.09±7.56%）, which was significantly higher than that in the D2 diluent group（7.96±2.42%, P＜0.05）, but there was no significant difference between the D1 diluent group and the D3 diluent group（22.56±9.36 %,P＞0.05）).The percentage of sperm with intact plasma membrane after thawing in D1 diluent group was the highest（35.80±11.30%）, which was significantly higher than that in the D2 diluent group（17.29±7.51%, P＜0.05）, but there was no significant difference between the D1 diluent group and the D3 diluent Group（33.26±6.30%,P＞0.05）.The percentage of intact acrosome spermatozoa after thawing was the highest in the D2 diluent group（66.02±6.40%）, which was significantly higher than that in the D1（50.43±6.12%） and D3 diluent groups（33.26±7.70%,P＜0.05）.The percentage of intact acrosome spermatozoa in the D1 diluent group was significantly higher than that in the D3 diluent group（P＜0.05）.The results indicated that Tris-citric acid-yolk dilution containing fructose could better protect frozen sperm of Black-bone sheep and improve movement, survival, plasma membrane integrity rate and acrosome integrity rate of sperm after thawing.