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1.
In the present study, methodology of gynogenetic induction in spotted halibut were developed and optimized; the sex ratio of putative meiotic gynogenetic diploids was determined using AFLP-based molecular sexing technique; the homozygosity of gynogenetic population was assessed as opposed to cultivated population. The results showed that high percentage of meiotic gynogenetic diploids were generated when the eggs fertilized with irradiated heterologous sea perch frozen sperm (30–50 mJ cm−2) were cold shocked in sea water of −1°C for 40–75 min at 5 min after fertilization. About 15,200 diploid gynogenetic larvae were achieved and they exhibited normal morphology similar to diploid control. The gynogenetic diploids were 100% female, which first confirmed the female homogamete (XX/XY) sex determination in spotted halibut. The genetic analysis showed that the average H O was, respectively, 0.404 and 0.724 in gynogenetic population and cultivated population, indicating an increase of homozygosity in gynogenetic population.  相似文献   

2.
This study reports the results on induced meiotic diploid gynogenesis and female homogametic nature in the Indian catfish, Heteropneustes fossilis. The eggs of H. fossilis were inseminated with conspecific sperm. The sperm suspension was diluted to 1 × 107 sperm mL−1 in Hanks balanced salt solution. Sperm were irradiated under UV light, with the exposure time ranging from 15 to 360 s (7500 ergs mm−2 for 60 s). The genetic inactivation of paternal chromosomes was confirmed by chromosome counting from the larval cells and the larvae also had a characteristic haploid syndrome. A typical ‘Hertwig effect’ in the yield of hatched larvae was observed with doses of UV exposure >75 s (9375 ergs mm2). Larvae resulting from sperm UV irradiated above 120 s (15 000 ergs mm2) were 100% haploids. Application of heat shock to the activated eggs was effective in suppressing the release of the second polar body (meiotic gynogenesis) and resulted in diploid gynogenetic larvae morphologically identical to those of the control. The best yield of diploid gynogens (49.3% with respect to the control) was found to be at 6 min after egg activation and the heat shock at 41 °C for a 1-min duration, at an ambient water temperature of 27 °C. A total of 113 diploid gynogenetic fry from seven different female fish were reared and subjected to sexing. All gynogenetic fish were female in contrast to the control, which had a mean sex ratio of 56.7% females (which was not significantly different from 50% female). From these results, the sex determination mechanism in H. fossilis was presumed to be female homogamety.  相似文献   

3.
Mass production of fast‐growing, all‐female muskellunge Esox rnasquinongy by gynogenesis requires optimized techniques of preventing second polar body extrusion. Heat, cold, and pressure shocks were evaluated for their efficiency of doubling the maternal genome. Muskellunge eggs (20–40 g) were activated with 1 mL ultraviolet (UV)‐irradiated (1,248 J/ m2) heterologous sperm of yellow perch Perca flavescens. Survival and ploidy (by flow cytometry) were determined during the eyed‐stage. Cold shocks of 1.3 × 1 C were applied at 5 or 20 min after gamete activation with water (time of initiation, TI) for a duration of 150 min and pressure shocks of 48,263 or 55,158 kPa (7,000 or 8,000 psi, respectively) at a TI of 4 rnin for 12 min. These shock treatments resulted in 43.7–95.0% diploid gynogens with corresponding yield of diploid gynogen (percent diploid gynogens × total percent survival) of 2.6–11.I%. Cold shocks applied at TI of 5 or 20 min after activation resulted in statistically similar percent survival, percent diploid gynogens, and yield of diploid gynogens. Heat shocks of 31 × 0.1C applied at a TI of 5 to 15 min for a duration of 5 min resulted in 4.8–21.1% diploid gynogens with yields of 0.1–0.4%. Cold and pressure shocks have better potential than heat shock for preventing the second polar body extrusion. Muskellunge eggs activated with UV‐irradiated yellow perch sperm, but not exposed to shock, resulted in 100% haploids with survival of 2.3–5.8%. No viable embryos were produced from the hybrid cross between muskellunge and yellow perch, thus, all diploids produced after the shock treatments were unambiguous meiotic gynogens. Muskellunge eggs fertilized with fresh muskellunge sperm (controls) showed 60.4–64.0% survival to the eyedstage and 100% diploidy. Considering that the sex‐determining mechanism in muskellunge follows the WZ female, ZZ male system, future efforts should be directed to test the efficiency of cold and pressure shocks for mass‐producing gynogenetic super female (WW) muskellunge.  相似文献   

4.
Turbot Scophthalmus maximus exhibits sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. In this paper, gynogenetic diploids of turbot were induced by activating egg development with ultraviolet (UV)-irradiated left-eyed flounder Paralichthys olivaceus sperm combined with cold shock to prevent extrusion of the second polar body. The results of UV irradiation experiments showed that survival, motility, and duration of activity of P. olivaceus sperm generally decreased with increase in UV dose. The typical Hertwig’s effect was observed after fertilized turbot eggs with UV-irradiated P. olivaceus sperm and the optimal UV dose for gynogenetic haploid production was 36,000 erg mm−2. At 15°C, appropriate timing of cold shock for retention of the second polar body in turbot eggs was at 6 min after fertilization. Results of different combinations of two shock temperatures (1 or 3°C) and four shock durations (15, 25, 35 or 45 min) at 6 min after fertilization demonstrated that shock of 25 min at 1°C gave the highest production of diploid gynogens (39.58% relative to its diploid control). The results of this study reveal that the use of UV-irradiated P. olivaceus sperm for activation of turbot eggs and cold shock for polar body retention is an effective method to produce gynogenetic offspring.  相似文献   

5.
A range of phenotypic characters were measured and their mean and coefficient of variation (CV) calculated in meiotic and mitotic gynogenetic individuals and normal diploids produced from the same full-sib family of Oreochromis niloticus (L.). The traits studied in all three groups were weight, length, various fin ray and scale counts, gonadosomatic index (GSI) and reproductive response. The study revealed two main trends in the majority of the traits studied in the gynogenetic fish: a decrease in the mean of each trait in the ranked order normal diploid > meiotic gynogenetic > mitotic gynogenetic, and an expansion of its phenotypic variation (CV) in the order normal diploid < meiotic gynogenetic < mitotic gynogenetic. This could be related to the levels of genetic homozygosity and a possible reduction in developmental homeostasis in the gynogenetics. The utility and potential of gynogenetic individuals in research and the use of the technique in the improvement of fish strains for aquaculture are discussed.  相似文献   

6.
为了解黄姑鱼(Nibea albiflora)异质雌核发育子代的基因纯合情况,利用微卫星标记(SSR)和扩增片段长度多态性标记(AFLP)对黄姑鱼异质雌核发育家系进行遗传鉴定和分析。结果显示:(1)雌核发育家系在4个SSR位点和5对AFLP引物组合扩增出的位点均未发现父本特异性等位条带,表明雌核发育体比率为100%。(2)用于遗传分析的7个SSR位点在雌核发育家系和正常交配家系中均未见完全纯合的情况,雌核发育家系7个SSR位点的平均纯合度为0.382,是正常交配家系(0.161)的2.37倍。雌核发育家系各个体的纯合位点数为0~6个,纯合位点所占比例为0~85.7%。(3)5对AFLP引物共扩增出182条清晰的扩增条带,其中有21条父本特异性条带和16条母本特异性条带。16条母本特异性条带中有7个条带在雌核发育家系中显著偏分离(P<0.05)。雌核发育家系和正常交配家系多态性条带比例分别为14.7%和20.3%。(4)雌核发育家系与母本的遗传相似度高于与正常交配家系的遗传相似度,正常交配家系同父本和母本的遗传距离大致相同。研究结果表明,黄姑鱼异质雌核发育二倍体家系的遗传纯合度显著高于正常交配家系,人工诱导雌核发育是促进基因纯合的一个有效途径,它不仅可以加速有利基因的纯合固定,还可以加速有害基因的淘汰,从而有效提高育种效率。  相似文献   

7.
The sex ratios and sex determination mechanism of gynogenetic diploids of ship sturgeon, Acipenser nudiventris, have been investigated to verify the possibilities of sex control by chromosome manipulation in this species. Meiotic gynogenesis was induced in a female ship sturgeon using cold shock after egg activation with UV-irradiated sperm of a male Siberian sturgeon. Microsatellite DNA analysis was applied for verification of uniparental inheritance in the gynogenetic diploid group of fish. All the analyzed gynogenetic diploids possessed only maternal genotype in the examined experimental group of fish. In this study, a minimum of two distinctly selected diagnostic loci in the offspring was used to confirm exclusively maternal contribution. Also, these fish were analyzed for sex diagnostic. Histological analysis of gonads from gynogenetic diploids, obtained from one family, showed 73.3 % of females and 27.7 % of males. The observed sex ratio has suggested that the ship sturgeon have a female heterogametic sex determination system. Gynogenesis in this species with female heterogametic sex determination system will have important role in breeding program and reclamation of its natural population to produce both female and male progeny, while this species has been introduced in the red list of IUCN (International Union for Conservation of Nature and Natural Resources).  相似文献   

8.
Abstract Optimum conditions for hydrostatic pressure treatment for duplication of chromosome set in gynogenetically activated fighting fish, Betta splendens (Regan), eggs were identified. Maximum survival of heterozygous gynogens was 50%, when 2·5-min-old eggs, after insemination with UV irradiated tilapia sperm, were pressure shocked at 7000psi for 6 min. The frequency (21%) of homozygous gynogenetic fry was high, when the 34min (post-insemination) old eggs, which were inseminated with tilapia sperm, were pressure shocked for 5 min. Sex ratio of gynogenetic progeny suggested that the mechanism of sex determination in this fish is homogametic female and heterogametic male.  相似文献   

9.
The bagrid catfish, Pseudobagrus ussuriensis, exhibits significant sexual dimorphism in growth rate and body size with males growing faster than females. Therefore, an all‐male culture can dramatically increase the output and profitability of fishery products. According to the monosex breeding route, super‐male individuals’ acquirement by XY male sex reversal and artificial gynogenesis is the key step. An effective protocol to induce meiotic gynogenesis using homologous sperms has been developed in this study. The most effective UV irradiation for sperm genetic inactivation was found to be at a distance of 20 cm with 66 μW/cm2 light intensity for 25 min. Optimal treatment for cold shock was 5 min post‐fertilization at 0‐4°C for 30 min, which gave the best survival rate of 13.65 ± 2.87%. The sex ratio in the meiotic gynogens showed a significant female‐biased deviation (< .05); thirty meiogynogens and their parents were further analysed using a male‐specific AFLP marker, of which only the male parent produced a male‐specific DNA band of 412 bp. These results indicated the female homogametic XX/XY sex determination system in P. ussuriensis. The optimization of a protocol for the successful induction of meiogynogenesis in the bagrid catfish lays the basis for all‐male production and is useful in ascertaining the genetic sex determination system in this promising aquaculture species.  相似文献   

10.
The honmoroko has been inferred to have an XX/XY sex determination system, but the parental genome can also affect the sex ratio of the offspring. The extent of parental effects on sex determination was examined by checking the sex ratios of F1 and F2 gynogenetic diploids and control diploids. Eleven gynogenetic broods from different females consisted of all or nearly all females, but eight broods showed a variable proportion of males (<50 %). One second-generation brood of gynogenetic diploids consisted wholly of females, but others produced some males. In crosses with a control diploid female, four males from a high-percentage male brood of gynogenetic diploids produced offspring with a balanced sex ratio. Sib-mating between a gynogenetic female and three gynogenetic males from the brood produced predominantly male progeny. These results suggest that there are at least four possible genotypes: genotypic female (XX), phenotypic female carrying a silent Y chromosome, genotypic male (XY), and genotypic supermale (YY). These inferences suggest that this fish has an XY system but a relatively high proportion of females possess a mutated, silent Y chromosome which does not lead to testis formation.  相似文献   

11.
Insemination of oocytes from six different salmon with irradiated heterospecific sperm combined with thermal shocks 10 min after activation successfully induced gynogenesis in pink salmon, Oncorhynchus gorbuscha (Walbaum). Biochemical genetic analysis and morphology of larvae con firmed their gynogenetic parentage, Thermal shocks of 26oC beginning 10 min after fertilization and lasting 15-30 min gave high yields of gynogenetic salmon (32% of normal diploid hatching, average of six females; 70% in oocytes from one female treated 20 min): shocks of 28oC lasting 10-16 min also gave high yields (2 5%), Shocks of 30oC lasting 6-12 min gave lesser yields (10%), Female donors varied significantly in the susceptibility of their oocytes to treatment (15-70% range of yield to average best treatment) but females did not interact significantly with heat shock treatment: different heat shocks had relatively the same effect on all females.  相似文献   

12.
Large yellow croaker, Pseudosciaena crocea, exhibit sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. We have established a protocol to produce all-female croaker P. crocea through induction of meiotic gynogenesis with homologous sperm. The first set of experiments investigated the ultra-violet (UV) irradiation on sperm motility and duration of sperm activity to determine the optimal UV dosage for genetic inactivation of sperm, yet retaining adequate motility for activation of eggs. Milt from several males was diluted 1:100 with Ringer’s solution and UV irradiated with doses ranging from 0–150 J cm−2. The results indicated that motility and duration of activity generally decreased with increased UV doses. At UV doses greater than 105 J cm−2, after fertilization, motility was <10% and fertilization rates were significantly lower. Highest hatching rate was obtained at 75 J cm−2. A second set of experiments was carried out to determine appropriate conditions of cold shock for retention of the 2nd polar body in P. crocea eggs after fertilization with UV-inactivated sperm by altering the timing, temperature and duration of shock. At 20°C, shock applied at 3 min after fertilization resulted in higher survival rate of larvae at 6 h after hatching. Results of different combinations of three shock temperatures (2°C, 3°C or 4°C) and five shock durations (4 min, 8 min, 12 min, 16 min or 20 min) at 3 min after fertilization demonstrated that shocks of 12 min gave highest production of diploid gynogens. Statistical analysis revealed that maximum production of diploid gynogens (44.55 ± 2.99%) were obtained at 3°C. The results of this study indicate that the use of UV-irradiated homologous sperm for activation of P. crocea eggs and cold shock for polar body retention is an effective method for producing gynogenetic offspring.  相似文献   

13.
Gynogenesis was induced using heterologous sperms in large-scale loach, Paramisgurnus dabryanus (Sauvage), in which a ZW/ZZ sex determination was previously proposed. Three microsatellite loci were used to monitor exclusive maternal inheritance of gynogenetic progenies. The results showed that high percentages of meiogynogens were produced at 4 min post-fertilization and mitogynogens were produced at 18 min post-fertilization by heat shocks, while meiotic gynogenesis was induced by cold shocks within a wide period and high heterozygosity was even observed in gynogens produced at 24 min post-fertilization. The sex ratios of the F1 progenies in three gynogenetic families were significantly deviated from 1:1 expectation with a female bias in two families and a male bias in one family (P < 0.05), and the other four gynogenetic families showed approximate 1:1 sex ratios. Moreover, the self-mating between gynogenetic F1 progenies and mating between gynogenetic F1 progenies and normal individuals produced all-female progenies or identical proportions of females and males. The data of sex ratios generally confirmed that the sex determination in large-scale loach was determined by the putative ZW/ZZ system, and the possible reasons causing the biased sex ratios are discussed.  相似文献   

14.
利用紫外线照射对七彩鲑(Salvelinus fontinalis)精子进行灭活处理,使其遗传物质失活,作为同源精子诱导源,与七彩鲑卵子进行受精,采用冷休克方法诱导雌核发育。结果表明:同源精子可以诱导七彩鲑的雌核发育,紫外照射强度为72 m J/cm2;距离为8 cm;冷休克温度为20℃。七彩鲑减数分裂雌核发育倍性测定以鸡(Gallus sp.)红细胞DNA含量(2.5 pg/N)为标准、七彩鲑普通育苗子一代为参照,采用流式细胞术和微卫星标记技术对30尾七彩鲑减数分裂雌核发育鱼苗进行分析,证明了雌核发育鱼苗为雌核发育二倍体。  相似文献   

15.
为了分析连锁群上不同位置的微卫星标记对鉴定牙鲆二倍体遗传特征的作用,实验以牙鲆选育基础群体为亲本,选择性腺发育良好的个体制备普通二倍体(ND),减数分裂雌核发育二倍体(MGD-1)、连续两代减数分裂雌核发育二倍体(MGD-2),并利用MGD-1发育达性成熟的雌鱼与同时诱导的性反转伪雄鱼交配制备近交二倍体(MGD1H)。从牙鲆遗传连锁图谱选择均匀分布于24个连锁群的72个微卫星标记,用4个MGD-1家系估计微卫星标记与着丝粒之间的相对距离。在假设无交叉干涉的情况下,17个标记位于着丝粒区域,19个标记位于连锁群中部,36个标记位于远着丝粒区域。分别选择上述区域的微卫星标记鉴定牙鲆4种二倍体的遗传特征。结果显示,4种二倍体的等位基因数(A)和多态信息含量(PIC)在不同区域变化范围较小,ND的A和PIC均为最高,MGD1H则表现为最低。随着标记与着丝粒之间距离的增加,4种二倍体的观测杂合度逐渐升高,纯合度逐渐降低。纯合个体比例在着丝粒区域最高,为8.8%~29.1%;在远着丝粒区域最低,为2.4%~23.2%。其中,MGD-1和MGD-2的变化幅度显著高于其余二倍体。结果表明,选择连锁群上不同位置的微卫星标记对鉴定牙鲆不同二倍体的遗传特征具有显著影响。  相似文献   

16.
The mandarin fish Siniperca chuatsi is a historically important aquaculture species in China and exhibits sexually dimorphic growth. However, sex determination of this fish remains unclear so far. In this study, we induced meiotic gynogenesis in S. chuatsi using irradiated heterologous sperm from spotted mandarin fish (Siniperca scherzeri) to uncover its mechanism of sex determination. Up to 7.52% diploid progeny were obtained among three gynogenetic families in this study. Molecular analysis of female and male donors and sampled young gynogens by seven microsatellite loci further confirmed no genetic contributions from the ‘father’ S. scherzeri. After 8 months of culture, external morphology of adult fish showed that all gynogens were cloned from their mothers. Gonads of the gynogenetic progeny were examined by histological observations and the sexing results showed that they were almost 100% females, strongly supporting an assumption of female homogamety in mandarin fish. By this study, we obtained pure lines of S. chuatsi and elucidated its genetic mechanism of sex determination, providing a basis for possible sex control breeding in this species.  相似文献   

17.
The Japanese ornamental (koi) carp is a popular decorative fish all over the world. In koi, clones have not yet been obtained, although production of fish with identical colour patterns could be of commercial interest. Mitotic gynogenetic progenies are essential for subsequent production of clones in fish. However, resulting late‐shocked progenies may be contaminated with meiotic gynogens from spontaneous suppression of the second meiotic division in eggs. In this study, microsatellite DNA markers were used to confirm mitotic gynogenetic origin of obtained late‐shocked progenies. Recombination rate (y) and mapping distance relative to centromere (M‐C) of 10 microsatellite loci were determined based on percentage of heterozygotes in meiotic gynogenetic progenies. The range of y varied from 0.01 to 0.96 and the M‐C map ranged from 0.5 to 48 cM. The mean value of y over the 10 loci was 0.481. Six loci, which had y 0.47 and higher, were used as markers in two late‐shocked gynogenetic progenies. Complete homozygosity was revealed at all six microsatellite loci indicating mitotic gynogenetic origin of analysed progenies.  相似文献   

18.
Muskellunge, Esox masquinongy, is an important recreational freshwater fish native to North America. Since muskellunge populations are often maintained through stocking efforts, advances in muskellunge reproductive technologies are of direct relevance to fishery enhancement. We evaluated the efficiency of inbreeding through induced meiotic diploid gynogenesis. Eggs from six female muskellunge were manually stripped and activated using ultra violet‐irradiated yellow perch, Perca flavescens, sperm. Hydrostatic pressure shocking regimes (48 263 kPa) were then applied to the eggs to prevent second polar body expulsion producing unambiguous meiotic gynogens. Six female dams and samples of 12–20 of their gynogenetic progeny were genotyped at seven polymorphic microsatellite loci. Chromosomal recombination frequencies of microsatellite loci based on retention of heterozygosity among gynogens ranged from 0.043 to 0.839 (0.576 ± 0.237). There were no statistical differences in recombination frequency among females at any of the loci. The average inbreeding coefficient (F‐value) ranged from 0.581 to 0.979, equivalent to three to fourteen generations of full‐sibling crosses respectively. The average F‐value overall was 0.712, equivalent to between five and six generations of full‐sibling crosses. Centromere map distances of the seven microsatellite loci ranged from 2.15 to 41.95 cM and meiotic gynogenesis was useful in eliminating heterozygosity at loci proximal to the centromere, but not distal. Since the age at maturity of female muskellunge is approximately 5 years, gynogenesis may pose an expeditious alternative to traditional breeding strategies for creation of homozygous pedigrees for some loci that may be outcrossed to introduce positive heterozygosity effects in the offspring.  相似文献   

19.
Gynogenetic diploids were induced in the loach, Misgurnus anguillicaudatus, by applying cold shock to the egg after fertilization with spermatozoa genetically inactivated by ultraviolet-ray irradiation, to examine their survival, growth and fertility. Eggs were obtained from orange phenotype loach (homozygous for a recessive demelanogenesis gene). Spermatozoa were obtained from black phenotype loach (homozygous for a dominant melanogenesis gene), and from common carp, Cyprinus carpio. All hybrids between loach and carp were inviably abnormal.Eggs fertilized with loach or carp spermatozoa irradiated at doses of 6000 and 12000 erg/mm 2 and then treated with cold water (1°C for one hour 4 and 5 min after fertilization developed into embryos with 61–78% survival. About 70–87% of newly-hatched fry were normal in appearance. Gynogenetic fish induced by both loach and carp spermatozoa showed lower rates of survival and growth than did the normal diploid controls. All gynogenetic fish from carp spermatozoa were female and 94.9% of the gynogenetic fish from loach spermatozoa were female. The second gynogenetic offsprings were produced from females selected from both gynogenetic groups.  相似文献   

20.
半滑舌鳎有效群体大小估计   总被引:1,自引:0,他引:1  
为了解半滑舌鳎当前的群体遗传结构,探究半滑舌鳎有效群体大小(Ne)发展趋势及现状,实验通过性别特异性分子标记随机鉴定了800尾半滑舌鳎的遗传性别,选取297个遗传雌性半滑舌鳎样本进行简化基因组测序(2b-RAD),获得了64 416个可用SNP标记,利用这些标记进行全基因组范围的连锁不平衡分析得到各染色体上连锁不平衡分布;根据标记间不同物理间距进行半滑舌鳎有效群体大小的估计,初步了解各历史世代下有效群体规模;通过选择0.1、0.2、0.5、1.0、2.0、5.0、10.0 Mb等7个不同染色体片段大小反映出半滑舌鳎群体遗传结构经过自然与人工共同选择的历史发展趋势。结果显示,半滑舌鳎有效群体大小随其连锁不平衡程度衰减而呈连续下降趋势,至2世代前其有效群体大小仅为29尾左右。  相似文献   

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