石鲽、牙鲆精子冷冻保存研究及其在人工杂交中的应用

Cryopreservation of stone flounder and Japanese flounder sperms and its application to the breeding

对石鲽(Kareius bicoloratus )和牙鲆(Paralichthys olivaceus)精子冷冻保存技术进行了研究,筛选到一个优良的稀释液MPRS,用MPRS冷冻保存石鲽、牙鲆精子,冻后成活率在70%以上。在石鲽冻精和牙鲆卵的杂交实验中,杂交组受精率是 28 4%±4 55(n=3),孵化率是 42 7%±7 35(n=3),胚胎发育正常;5 000尾杂交鱼苗有102尾成功度过变态期。在牙鲆冻精和大菱鲆卵的杂交实验中,杂交组的受精率只有2 7%,部分受精卵虽能孵化出膜,但仔鱼全为畸形。这两个杂交实验,特别是石鲽冻精和牙鲆卵的杂交实验,证实了冷冻保存的精子完全可以应用在杂交育种工作中。

In the present study, cryopreservation of Kareius bicoloratus and Paralichthys olivaceus sperms was first investigated. Effect of extenders on the motility score of frozen-thawed sperm was examined. A good extender MPRS (modified plaice ringer solution) was selected and motility of frozen-thawed sperm was more than 70%. In order to confirm that the frozen stone flounder sperm has the fertilization ability with the Japanese flounder egg and the frozen stone flounder can be applied in the hybridization breeding, the hybridization trial (Japanese flounder ♀× stone flounder ♂) was done. The Japanese flounder ♀× stone flounder ♂ hybridized embryo hatched 70~75 h after fertilization and no significant difference was found between hybridized group and control (Japanese flounder ♀×Japanese flounder♂).In order to get a hybridization species,the hybridization trial between turbot egg and Japanese flounder frozen sperm was performed. But the hatched hybridized fry were all abnormal because of the chromosome differences.