虾夷扇贝Patinopecten yessoensis (Jay)和仿刺参Apostichopus japonicus (Seienka)多倍体诱导研究

IOCAS-IR > 海洋环流与波动重点实验室

	虾夷扇贝Patinopecten yessoensis (Jay)和仿刺参Apostichopus japonicus (Seienka)多倍体诱导研究
	常亚青
学位类型	博士
	2000
学位授予单位	中国科学院海洋研究所
学位授予地点	中国科学院海洋研究所
学位专业	海洋生物学
关键词	虾夷扇贝仿刺参化学诱导药物多倍体苗种
摘要	本文采用化学方法诱导了虾夷扇贝Patinopecten yessoensis (Jay)和仿刺参Apostichopus japonicus (Selenka)的多倍体。采用6－二甲基氨基嘌呤（6－dimethylaminopurine）抑制虾夷扇贝极体释放，研究了诱导三倍体的实验条件、技术工艺以及三倍体虾夷扇贝幼虫的生长和培育方法。结果表明，使用6－二甲基氨基嘌呤（6－DMAP）可诱导17.3%-100.0%的三倍体虾夷扇贝面盘幼虫。筛选并优化了生产中操作简便、高效、成活率高的诱导条件，并研究出在扇贝三倍体诱导中批量获得受精卵、处理及培育的技术工艺，对扇贝三倍体的规模化诱导及培育等生产性应用具有较高的参考价值。报道了采用细胞松弛素B（cytochalasin B）、秋水仙素（colchicine）、6－DMAP以及咖啡因（coffeine）等药物抑制虾夷扇贝第一极体（PB_1）释放、PB_1和第二极体（PB_2）释放以及抑制第一次卵裂等方法诱导四倍体的结果。结果表明，CB、6－DMAP和秋水仙素抑制扇贝第一次有丝分裂诱发四倍体的比例低于25%，而采用CB抑制PB_1可有效地诱导产生四倍体，从授精后42min提前到15-22min开始处理，抑制PB_1的放出有助于提高四倍体的比例，在12 ℃，处理开始和终上时间分别在授精后20－22min和60－62min时，面盘幼虫四倍体率最高，为56.5%。对CB处理抑制受精卵PB_1释放的处理组胚胎的染色体分离状况进行了观察研究。对照组受精卵具有19条四分体染色体，经过减数分裂I期（meiosis I）和减数分裂II期（meiosis II），放出PB_1和PB_2，受精卵的发育具有不同步性。处理组受精卵在第二次减数分裂中出现了“三级分离“、“联合二级分离“和“独立二级分离“等特殊的分离类型，初步分析了CB抑制第一极体放出的机理。对三倍体虾夷扇贝的繁殖潜力和卵子的大小进行了观察研究，三倍体扇贝具一定的繁殖能力，三倍体雌贝平均产卵3.26 * 10~6个，而相同壳长二倍体贝为1.45 * 10~7，三倍体产卵量仅为二倍体产卵量的22.5%。三倍体卵子产出后，形状不规则，卵子平均直径为87.25μm，比二倍体大11.7%，卵子体积比二倍体大39.4%。利用CB抑制第一极体释放诱导了虾夷扇贝的四倍体，诱导率为41.5%。首次报道了仿刺参Apostichopus japanicus (Selenka)多倍体诱导的结果。采用紫外线照射的海水成功地使海参分别单独产卵、排精，从而准确地控制了海参的人工授精后处理的时间。采用0.2、0.4mg/L CB 抑制受精卵第一极体释放以及10、20、30和40mg/L 的6－DMAP 分别抑制PB_1、PB_2放出的方法诱导了仿刺参的多倍体。研究了诱导的药物浓度、处理时间以及处理开始时间等，同时对幼体的成活率等进行了探讨。结果表明，2种药物均可诱导仿刺参产生三倍体和四倍体，从效果上看，采用CB抑制PB_1诱导，到达小耳幼体时，可产生9.7%-21.3%的四倍体。6－DMAP抑制PB_2放出诱导三倍体，三倍体诱导率介于7.5%-25.8%之间。
其他摘要	Polyploidy Ezo scallop Patinopecten yessoensis (Jay) and Japanese sea cucumber Apostichopus japonicus (Selenka), which are the mainly cultured scallop and echinopermata species in China, were induced with chemicals. Triploid were induced by blocking the releasing of polar body with 6-dimethylaminopurine (6-DMAP) on the scallop. The induction methods and larvae cultivation, veliger growth were also reported. It indicated that 6-DMAP is the proper chemical inducer on this scallop triploid induction, and the triploid juvenile ratio were 17.3%～100.0%. For the large scale ploidy manipulation in scallop, it also give the methods how to get mass zygotes and the triploidy larvae cultivation. Tetraploid induction also reported in this scallop. Tetraploid were induced with the inhibition of polar body I, and both polar body I and polar body II extrusion, and the first cell cleavage, by the using of cytochalasin-B(CB), colchocine, 6-dimethylaminopurine (6-DMAP) and coffeine respectively or combined. The results shows that the tetraploidy percentage of scallop larvae were low than 25% with the first cell cleavage inhibiting by CB, 6-DMAP and colchicine. Tetraploid can be carried out by the first polar body inhibiting with CB. The yield of tetraploid increased with the shocking initial time from 42 minutes postfertilization to 15-22. The highest tetraploidy percentage was 56.5% at veliger larvae shocked begin at 20-22 minutes postfertilization to 60-62 minute. Fertilized eggs of scallop was treated with cytochalasin B to block polar body I, and the chromosomes segregation of control and shocked groups were analysis. The results shows that the fertilized egg in the control group has 4 suits of 19 chromosomes and release polar body I and polar body II after meiosis I and II. The development of fertilized egg was asynchronous. The several special mode of chromosomes segregation, tripolar segregation, united bipolar segregation and separated bipolar segregation were observed in the treated groups during meiosis II. The mechanism of polar body I inhibition with CB were also analyzed preliminarily. The triploidy individual spawning capability and eggs size were reported. The induced spawning number of one triploidy scallop was 3.26 * 10~6, it was 22.5% of the control. The eggs shape spawned by triploid were anomaly, it's diameter is 87.25μm, 11.7% larger in diameter and 39.4% in volume than control, respectively. The highest yield ratio of tetraploid by CB shocking was 41.5%. The polyploidy sea cucumber Apostichopus japonicus (Selenka) were induced. It give a method with UV irradiated sea water to get the eggs and sperm separately, in order to control the beginning time of shocking. The shocked chemicals were CB and 6-*DMAP, inhibit polar body I with 0.2 and 0.4mg/L CB density, and inhibit the polar body I and II respectively with 10,20,30,40mg/L 6-DMAP. It shows that polyploid can be induced by the two chemicals, with 9.7%-21.3% tetraploid percentage shocked by CB and 7.5%-25.8% triploid percent shoked by 6-DMAP.
页数	92
语种	中文
文献类型	学位论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/1499
专题	海洋环流与波动重点实验室
推荐引用方式 GB/T 7714	常亚青. 虾夷扇贝Patinopecten yessoensis (Jay)和仿刺参Apostichopus japonicus (Seienka)多倍体诱导研究[D]. 中国科学院海洋研究所. 中国科学院海洋研究所,2000.

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