人工雌核发育诱导太平洋牡蛎四倍体的研究

The induce on tetraploid of Pacific oyster by artifical gynogenesis

通过人工雌核发育诱导太平洋牡蛎(Crassostrea gigas)四倍体。精子遗传失活采用紫外线照射法,紫外线照射时间1 min、紫外线照射剂量1 500-W.cm-2。诱导雌核发育四倍体用6-二甲基氨基嘌呤(6-DMAP),选用L9(34)设计,进行三因素三水平的正交试验。诱导持续时间(A)设35,45,55 min;处理起始时间(B)10,15,20 min;6-DMAP浓度(C)设300,450,600μmol.L-1等三水平,试验重复二次。解剖法获取精卵,人工授精。染色体倍性检查采用染色体计数法。根据正交试验直观分析结果,得出诱导太平洋牡蛎四倍体各因素的最优水平组合:处理起始时间15 min、6-DMAP浓度450μmo.L-1、诱导持续时间45 min;三因素主次顺序:6-DMAP浓度→处理起始时间→诱导持续时间。

It was reported that the methods of artificial gynogenesis induced tetraploid in the Pacific oysten （Crassostrea gigas）. Ultraviolet light-irradiated was 1 500 μW·cm^-2 and the time was 1 min. 6-dimethyla minopurine （6-DMAP）was used to restrain the release of the first and second polar body during meiosis Ⅰ and meiosis Ⅱ. L9 （3^4） design was selected to engage in orthogonal experiment of three factors and three levels： the three levels of inducing duration （A） were 35, 45, 55 min, respectively; the beginning of inducing occasions（the time after sperms and eggs mixed）（B）were 10, 15, 20 min, respectively; the concentration of 6-DMAP （C） were 300, 450, 600μmol·L^-1, respectively. The experiments were repeated twice. The method of chromosome counts was used to examine chromosome ploidy. The result showed the optimum conditions： inducing duration was 45 min, the beginning of inducing occasion was 15 min, 6-DMAP concentration was 450μmol·L^-1. The sequence of decisive factors was 6-DMAP concentration→the beginning of inducing occasions→inducing duration.