本研究采用紫外线灭活的真鲷(Pagrus major)精子激活牙鲆(Paralichthys olivaceus)卵子,经过静水压机处理诱导有丝分裂雌核发育二倍体。优选施压起始时间、持续时间和压力大小3个方面参数,获得最佳参数组合为：起始时间60 min、持续时间6 min、施加压力650 kg/cm2。在此条件下的受精率最高为67.80%,孵化率54.23%,与其他处理组间达显著性差异(P<0.05)。用流式细胞仪检测未经静水压处理的胚胎,其 DNA 含量约为普通二倍体的1/2,即单倍体；检测有丝分裂雌核发育胚胎,其DNA含量与普通二倍体大体一致。结果表明,应用该方法可成功诱导获得牙鲆有丝分裂雌核发育二倍体。%Mitotic gynogenetic diploid Japanese flounder, Paralichthys olivaceus were produced by activating eggs with UV irradiated sperm of red sea bream (Pagrus major), followed by hydrostatic pressure treatment to block the first mitotic division. The initiation time, duration time and pressure value of hydrostatic pressure were opti-mized in this study. The results showed that the treatment with hydrostatic pressure (650 kg/cm2) for 6 min starting 60 min after fertilization was the optimal parameters combination. Under this condition the fertilization rate and hatching rate at this scenario were 67.80%and 54.23%, respectively, which were significantly higher than that of all the other treatment groups(P<0.05). The embryo not treated by hydrostatic pressure (haploid) were identified via flow cytometer, and its relative DNA content was about half that of normal diploid. The mitotic gynogenetic embryo had the same DNA relative content with normal diploid. The analysis results showed that mitotic gynoge-netic diploids could be successfully prepared.
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