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采用抑制第二极体排放的方式对15℃孵育的中华绒螯蟹自然受精卵人工诱导三倍体,通过抑制第一次卵裂对18℃孵育的自然受精卵人工诱导四倍体,应用流式细胞仪进行倍性鉴定.分别使用CB, 6-DMAP和KCl三种试剂对受精卵处理,发育至囊胚期检测,获得的最高三倍体诱导率依次是49 1%, 51 7%和77 5%,获得的最高四倍体诱导率依次是50 3%, 54 9%和79 8%.使用KCl试剂对抱卵蟹诱导处理,孵出状幼体检测,最高三倍体诱导率为85 3%,最高四倍体诱导率为27 3%.克服了以往三倍体诱导中离体培养的困难,首次获得了中华绒螯蟹三倍体状幼体,并极大提高了状幼体阶段四倍体诱导率.
Triploid Chinese mitten crab (Eriocheir sinensis) natural fertilized eggs incubated at 15 ℃ were artificially induced by means of inhibition of the discharge of the second polar body. The tetraploid was induced by inhibiting the first cleavage of natural fertilized eggs incubated at 18 ℃. Cytometry was used to identify the ploidy.The fertilized eggs were treated with CB, 6-DMAP and KCl, respectively, and developed to the blastocyst stage. The highest triploid inducibility rates were 49 1%, 51 7% and The highest rate of tetraploid induction was 50 3%, 54 9% and 79 8%, respectively.Using KCl reagent to induce oviposition, the highest triploid induction rate was 85 3%, and the highest tetraploid induction rate was 27 3% .Thirdly, it overcomes the difficulty of in vitro culture in triploid induction and obtained the first triploid trisomy of Eriocheir sinensis, Phase tetraploid induction rate.