Artificial induction of mito-gynogenetic diploids in large yellow croaker (Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic (endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea. In haploid control groups, the development of eggs was activated with ultraviolet radiated semen. All fry presented typical haploid syndrome in the haploid control groups, and were verified as haploids using cytometry. After hydrostatic pressure treatment, morphologically normal fry reappeared at different frequencies according to the intensity and time of pressure shock. Fry with normal appearance in the pressure treated groups were verified as gynogenetic double haploids (GDHs), containing only one allele from the female parent at all four diagnostic microsatellite loci. For a fixed duration of 3 min, the optimal intensity of blocking the first mitosis was determined to be 40 Mpa, which was similar to that of blocking the second meiosis. There was a “window” of starting time, from 36.1 min to 38.1 min post-insemination at 25.0±1.0°C, within which the production of GDHs was not significantly different. Maximum production of morphologically normal fries, 9.36%±2.97% of developed eggs, was found when the eggs were shocked with hydrostatic pressure at 40 Mpa for 3 min, starting from 38.1 min post insemination at 25.0±1.0°C.     

棘头梅童鱼的细胞遗传学特征和X1X1X2X2/X1X2Y性染色体系统

我们通过荧光染色、自身基因组原位杂交（Self-GISH）和多色荧光原位杂交（FISH），首次研究了棘头梅童鱼（Richardson，1844）的核型特征。雌性核型有24对端部着丝粒染色体（2n=48a，NF=48），而雄性核型包含22对端部着丝粒染色体，2条端部着丝粒染色体单体和1条中间着丝粒染色体（2n=1m+46a，NF=48）。雌性和雄性核型之间的差异表明，棘头梅童鱼的性染色体系统为X₁X₁X₂X₂/X₁X₂Y型，其中Y为雄性中特有的中间着丝粒染色体。三色FISH结果显示，5S rDNA和18S rDNA位点定位在最大的端着丝粒染色体（X₁）以及Y染色体的短臂；X₁染色体上有一个特异的臂间端粒信号（ITS），与5S rDNA位点部分重叠。Self-GISH结果显示，在推定的性染色体DNA重复序列聚集。根据实验结果我们提出关于棘头梅童鱼Y染色体起源的假说：Y染色体起源于祖先核型（2n=48a）中的两条端部着丝染色体融合，并且在此过程中伴随着片段缺失。本研究首次在石首鱼科中描述了异形的性染色体，将为其他石首鱼的性染色体研究提供线索。     

Molecular cytogenetic of the Amoy croaker,A rgyrosomus amoyensis(Teleostei,Sciaenidae)

The family Sciaenidae is remarkable for its species richness and economic importance. However, the cytogenetic data available in this fish group are still limited, especially those obtained using fl uorescence in situ hybridization(FISH). In the present study, the chromosome characteristics of a sciaenid species, Argyrosomus amoyensis, were examined with several cytogenetic methods, including dual-FISH with 18 S and 5 S rDNA probes, and a self-genomic in situ hybridization procedure(Self-GISH). The karyotype of A. amoyensis comprised 2 n=48 acrocentric chromosomes. A single pair of nucleolar organizer regions(NORs) was located at the proximal position of chromosome 1, which was positive for silver nitrate impregnation(AgNO_3) staining and denaturation-propidium iodide(DPI) staining but negative for Giemsa staining and 4',6-diamidino-2-phenylindole(DAPI) staining, and was confi rmed by FISH with 18 S rDNA probes. The 5 S rDNA sites were located at the centromeric region of chromosome 3. Telomeric FISH signals were detected at all chromosome ends with dif ferent intensities, but internal telomeric sequences(ITSs) were not found. Self-GISH resulted in strong signals distributed at the centromeric regions of all chromosomes. C-banding revealed not only centromeric heterochromatin, but also heterochromatin that located on NORs, in interstitial and distal telomeric regions of specifi c chromosomes. These results suggest that the karyotype of Amoy croaker was relatively conserved and primitive. By comparison with the reported cytogenetic data of other sciaenids, it can be deduced that although the karyotypic macrostructure and chromosomal localization of 18 S rDNA are conserved, the distribution of 5 S rDNA varies dynamically among sciaenid species. Thus, the 5 S rDNA sites may have different evolutionary dynamics in relation to other chromosomal regions, and have the potential to be ef fective cytotaxonomic markers in Sciaenidae.     

杂色鲍与盘鲍杂交F1的早期生物学表现

比较研究杂色鲍(Haliotis diversicolor)(S)与盘鲍(H.discus discus)(J)正反交F1及其亲本自繁后代的卵裂速度和孵化速度,以及杂色鲍与盘鲍杂交F<,1>及杂色鲍自繁幼体的成活率.在22～23℃下,杂色鲍自繁后代的早期发育速度快于盘鲍自繁后代.杂色鲍和盘鲍2细胞期分别为授精后45.3...     

九孔鲍精子短期保存技术研究

应用单因素试验和三因素正交试验研究了温度、二甲基亚砜(DMSO)浓度及精子密度等因素对九孔鲍(Haliotis diversicolor supertextexta)精子短期保存的影响。单因素试验结果表明: DMSO、维生素C、甘油、葡萄糖等添加剂中仅5%和7. 5% DMSO有利于精子活力的保持; 3个试验温度中28℃精子活力下降速度最快，其次为1℃, 最慢是5℃; 稀释度越高精子活力下降速度越快。正交试验中, 九孔鲍精液分别在25个保存条件下保存96 h, 保存期间每隔一段时间取样观察精子活力。25 个处理的活力变化数据用生长函数拟合, 根据拟合方程计算出精子活力半衰期, 并对结果进行直观分析及方差分析。结果表明, 较理想的精子保存条件是: 温度为7℃, DMSO质量分数为3%, 精子密度为7.5×10⁸个/mL; 各因素对精子活力半衰期的影响按精子密度、温度、DMSO密度顺序递减; 精子密度对精子活力半衰期有显著影响(P<0.05) , 温度及DMSO浓度的影响不显著(P>0.05) 。     

Artificial induction of mito-gynogenetic diploids in large yellow croaker (Pseudosciaena crocea) by hydrostatic pressure

The present study investigated conditions for inducing mito-gynogenetic(endomitosis) diploids by hydrostatic pressure in the large yellow croaker Pseudosciaena crocea.In haploid control groups,the development of eggs was activated with ultraviolet radiated semen.All fry presented typical haploid syndrome in the haploid control groups,and were verified as haploids using cytometry.After hydrostatic pressure treatment,morphologically normal fry reappeared at different frequencies according to the intensity and...     

九孔鲍精子短期保存技术研究

应用单因素试验和三因素正交试验研究了温度、二甲基亚砜(DMSO)浓度及精子密度等因素对九孔鲍(Haliotis diversicolor supertexta)精子短期保存的影响。单因素试验结果表明:DMSO、维生素C、甘油、葡萄糖等添加剂中仅5%和7.5%DMSO有利于精子活力的保持;3个试验温度中28℃精子活力下降速度最快,其次为1℃,最慢是5℃;稀释度越高精子活力下降速度越快。正交试验中,九孔鲍精液分别在25个保存条件下保存96 h,保存期间每隔一段时间取样观察精子活力。25个处理的活力变化数据用生长函数拟合,根据拟合方程计算出精子活力半衰期,并对结果进行直观分析及方差分析。结果表明,较理想的精子保存条件是:温度为7℃,DMSO质量分数为3%,精子密度为7.5×108个/mL;各因素对精子活力半衰期的影响按精子密度、温度、DMSO密度顺序递减;精子密度对精子活力半衰期有显著影响(P<0.05),温度及DMSO浓度的影响不显著(P>0.05)。