中华绒螯蟹多倍体诱导技术改进
Improvement of polyploidy induction in Eriocheir sinensis
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摘要: 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,孵出潘状幼体检测,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潘状幼体,并极大提高了潘状幼体阶段四倍体诱导率.Abstract: A incubating temperature of 15℃ was used to induce triploidy in Eriocheir sisnensis through inhibition of the release of polar bodyⅡ,and 18℃ was used to induce tetraploidy through inhibition of the first cleavage.Flow cytometry was used to identify the ploidy in different developmental stages.For induction of triploidy in fertilized eggs in vitro,the highest induction rate observed in blastulaby cytochalasin B,6-DMAP and KCl was 49.1%,51.7% and 77.5%,respectively.In KCl treatment of pregnant crabs with fertilized eggs,the highest triploid induction rate observed in the zoea was 85.3%.For induction of tetraploidy,the highest induction rate observed in the blastula by cytochaslasin B,6-DMAP and KCl were 50.3%,54.9% and 79.8% respectively.In the KCl treamtent of pregnant crabs with fertilized eggs,the highest induction rate in zoea was 27.3%.Throughth is study such difficulty as in vitro culture is overcome.Triploid zoea Eriocheir sisnensis was developed for the first tmie.The induction rate of tetraploid zoea was also greatly miproved.
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Key words:
- Eriocheir sinensis /
- triploid /
- tetraploid /
- zoea
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