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1.
Induction of triploidy in grass carp was accomplished by means of thermal shocks to eggs shortly after fertilization. Triploidy occurred most often with cold shocks at 5–7°C and at durations of 25–30 min starting 2.0–4.5 min after fertilization. Estimated percent triploid ranged from 50 to 100% on five occasions. With one exception, cold shocks of 5–7°C for less than 25 min did not induce triploidy, and cold shock durations of 30 min or longer generally resulted in 100% mortality. A heat shock of 40°C for 1 min, 4.75 min after activation, was the only heat treatment which produced triploidy (8%) with 81% surviving to the blastula stage. Fertilized eggs immersed in a solution of cytochalasin B (10 mg/l, 0.1% DMSO) for 10 min, 12 min after activation, resulted in 54% of the eggs surviving to the blastula stage with none found to be triploid. 相似文献
2.
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
3.
采用紫外线照射灭活精子,经冷休克诱导染色体加倍的方法对翘嘴鳜(Siniperca chuatsi)进行人工诱导雌核发育育种。灭活翘嘴鳜精子,紫外线照射剂量80 mJ/cm2是最佳的辐射剂量。冷休克结果表明,在0～3℃冰水下,冷休克起始时间为授精后3～7 min均有效,以5 min为最适;冷休克持续时间10-50 min均有效,以30 min最佳。综合以上两项因素,授精5 min后,休克时间持续30 min组的原肠胚存活率和孵化率均为最高,与其他处理组差异显著(P<0.05),此时雌核发育率最高,达到4.72%。 相似文献
4.
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
5.
Methods were determined for the induction of diploid gynogenesis in the sole, Solea solea (L.), in order to investigate the sex-determining mechanism. The experiments utilized gametes obtained by dissection of fish caught at sea. Activation of eggs with UV-irradiated sole sperm was not feasible but development was initiated by exposure to halibut, Hippoglossus hippoglossus (L.), sperm. These embryos displayed the typical characteristics of haploids and few survived beyond hatching. High levels (> 90%) of diploidy were induced in such eggs by subjecting them to a cold shock (2°C or 4°C) for 1-2 h about 10-15 min after activation at 12°C. The resultant embryos were indistinguishable from those developing from normally fertilized eggs and were considered to be diploid gynogenomes. Fifteen of these were reared to a length of 5-10 cm by which time gonad differentiation had been initiated. Both sexes were represented, indicating that sex in this species is not determined by a simple XX-XY system with female homogamy. 相似文献
6.
Jian-He Xu Feng You Wei Sun Bin-Lun Yan Pei-Jun Zhang Bi-Xiang Jing 《Aquaculture International》2008,16(6):623-634
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
7.
Shmuel Rothbard William L. Shelton Zeev Kulikovsky Israel Rubinshtein Yair Hagani Boaz Moav 《Aquaculture International》1997,5(1):51-64
The mollusc-eating black carp (Mylopharyngodon piceus) has economic and health-care potential for biological control of nuisance aquatic molluscs. The present study investigates
the production of gynogenetic-monosex and triploid-sterile populations of black carp. The goal was to provide a method which
would eliminate unwanted biological and environmental impacts of introducing this exotic species into areas with nuisance
mollusc infestation. Meiotic gynogenesis was induced by inseminating black carp eggs with UV-irradiated (800 Jm−2) sperm of common carp (Cyprinus carpio) or Japanese ornamental (koi) carp. Diploidy was restored through retention of the second polar body (2PB), by shocking activated
eggs at 1–8 min post-fertilization (embryological age of 0.07–0.57τ0, a parameter defined by the cell cycle duration) at 1 min intervals, with heat-shocks (41.0±1.0 °C for 1 min) or pressure-shocks
(7500–7600 psi for 1.5 min). Highest survival was found when embryos were heat-shocked 1.5–4.5 min post-fertilization (0.10–0.25τ0). The highest survival of free-swimming larvae from pressure-shocked eggs, was achieved at 7500 psi at 1–2 min post-fertilization
(0.08–0.16τ0). Triploidy was induced by retention of 2PB following normal fertilization. Batches of 30 000 eggs were fertilized with intact
sperm and pressure-shocked (6000–8500 psi for 1.5 min) 2 min post-fertilization (0.15–0.16τ0). The highest survival of triploid swim-up larvae was 5.1% in eggs shocked with 7500 psi. In random samples of individual
larvae taken from each treatment, triploidy was analysed by cytofluorometry of the cellular DNA content. In DNA analysis performed
in fingerlings (N≥15), 50% of the fish were triploids. 相似文献
8.
Mary Ann Garcia-Abiado Jacques Rinchard Konrad Dabrowski 《Journal of the World Aquaculture Society》2001,32(2):195-201
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
9.
Aquarium-held bigmouth buffalo Ictiobus cyprinellus (Valenciennes) were fed the blue-green alga Spirulina platensis (Nordstedt) at a daily rate of 29 g dry weight/kg body weight for 28 days. Daily growth was 14 g/kg body weight, and food conversion was 2.0. Tilapia aurea (Steindachner) also was fed this blue-green alga; growth and food conversion were similar to those found for bigmouth buffalo. Fresh Spirogyra sp. fed to grass carp Ctenopharyngodon idella Valenciennes resulted in poor growth, and food conversion was about 10. 相似文献
10.
雌核发育草鱼群体及两个普通草鱼群体的微卫星遗传分析 总被引：1，自引：1，他引：0
以草鱼(Ctenopharyngodon idellus)养殖群体(YZ)为母本,以紫外线灭活的鲤鱼精子激活草鱼卵子,冷休克抑制第二极体排出的方法诱导获得异精雌核发育草鱼群体(CH)。利用12对微卫星引物对YZ群体、YS (扬州广陵长江系家鱼原种场引进草鱼群体)群体、CH群体进行PCR扩增并分析,共检测出194个等位基因,其中75.8个有效等位基因。YZ群体、YS群体、CH群体的平均等位基因数依次为13.0、12.6、4.7;平均有效等位基因数依次是7.7、6.6、2.3;平均期望杂合度依次为0.87、0.82、0.56;平均多态信息含量依次为0.84、0.79、0.49。从每个个体在微卫星位点的纯合率看,YZ群体中个体的纯合度在0.00～0.33, YS群体r中个体的纯合度在0.00～0.42, CH群体中个体的纯合度在0.42～0.92。这表明与YZ群体和YS群体相比,CH群体的遗传多样性显著下降,并且在每个位点的纯合率CH群体均高于普通草鱼群体,表明人工诱导减数雌核发育可加速草鱼大多数基因位点的纯合,是快速建立高纯品系的有效手段。同时,本研究筛选并利用微卫星位点组合建立了雌核发育草鱼子代不同家系及其母本亲缘关系的简易、高效鉴别技术,旨在为雌核发育草鱼标记辅助育种打下基础。 相似文献
11.
雌核发育草鱼的遗传结构分析和微卫星鉴别方法的建立 总被引：2，自引：0，他引：2
采用微卫星标记检测草鱼群体的遗传多样性,并根据纯合度的变化建立了雌核发育草鱼的鉴别技术。结果显示,8个位点共扩增出33个等位基因;在普通草鱼中,平均纯合度和PIC分别为0.203 1和0.552 8;在雌核发育群体中,则为0.716 1和0.357 2;2个群体间遗传相似度为0.873 3。其中,5个位点在雌核发育草鱼中纯合度明显提高,雌核发育草鱼在这5个位点的扩增总条带数为5~7个,普通草鱼则为8~10个,由此可100%区分2个草鱼群体。通过概率计算,理论鉴别概率达到99.92%。研究表明,雌核发育技术对草鱼的群体遗传结构改变较大,是快速建立纯系、固定优良性状的有效手段;根据群体遗传纯合度的改变、扩增条带数目差异,应用多态性微卫星分子标记可以简单、有效地区分雌核发育群体(或与之相似的高度近交群体)与普通群体。 相似文献
12.
Reciprocal hybrids between tench Tinca tinca (L.) and carp Cyprinus carpio L., and tench and bream Abramis brama (L.) were produced artificially. The survival of all these hybrids during embryogenesis was quite high. The highest survival
rate (over 60%) at the eyed stage was observed for tench and bream hybrids (both sex combinations). The hatching rates of
these hybrids were also over 60%. The number of larvae with some abnormalities (i.e. deformed body) was low. In contrast,
the hatching rates of tench and carp hybrids were very low (0.2%). From over 1000 fertilized eggs, only three specimens started
swimming, and only one specimen survived to juvenile stage. Embryos of hybrids and their parental species differ in morphological
features. These differences were also visible in the juveniles. Body parameters of juvenile hybrids produced from three species
had intermediate values in comparison to parental fish. 相似文献
13.
John Samuel Godwin Christopher Arunachalam Ganesan Murugesan Natarajapillai Sukumaran 《Aquaculture Research》2010,42(1):129-138
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
14.
Some species of fish are more tolerant of rotenone, a commonly used non-specific piscicide, than others. This species-specific tolerance to rotenone has been thought to be associated with the uptake and the efficiency at which the chemical is detoxified. However, rotenone stimulates oxidative stress and superoxides, which are also toxic. Understanding the modes in which fish physiologically respond to rotenone is important in developing improved protocols for its application in controlling aquatic nuisance species. Using a molecular approach, we investigated the physiological and molecular mechanisms of rotenone resistance. Species-specific responses were observed when rotenone-sensitive silver, Hypophthalmichthys molitrix, and both rotenone-resistant bighead carp, Hypophthalmichthys nobilis, and bigmouth buffalo, Ictiobus cyprinellus, were exposed to rotenone. Rotenone levels in plasma were highest 90?min after exposure in both silver carp and bigmouth buffalo, but bigmouth buffalo tolerated over twice the burden (ng?mL(-1)?g(-1)) than silver carp. Expression of genes related with detoxification (cyp1a and gst) increased in silver carp, but either decreased or remained the same in bighead carp. Genes linked with oxidative stress in the cytosol (gpx, cat and sod1) and hsp70 increased only in silver carp after a 6-h exposure. Expression of genes associated with oxidative stress in the mitochondria (sod2 and ucp2) differed between silver carp and bighead carp. Expression of sod2 changed minimally in bighead carp, but expression of ucp2 linearly increased to nearly 85-fold of the level prior to exposure. Expression of sod2 and ucp2 did not change until 6?h in silver carp. Use of sod1 and sod2 to combat oxidative stress results in hydrogen peroxide production, while use of ucp2 produces nitric oxide, a chemical known to inhibit apoptosis. We conclude that the mechanism at which a fish handles oxidative stress plays an important role in the tolerance to rotenone. 相似文献
15.
Abstract. The use of a large-meshed seine net (aperture 2–5 cm) in enclosed compounds was an effective method for the capture of large grass carp, Ctenopha-ryngodon idella (Val.), and tench, Tinea tinea L., with mean fork lengths of 42–9 cm and 38–3 cm and mean weights of 1–53 kg and 1–10 kg respectively. Calculated population estimates using a constant-effort removal technique showed a high degree of accuracy for grass carp, when the absolute index of population size was checked after completion of netting with an application of a commercially available rotenone formulation. It is suggested that this was so because of the large proportion (>90%) of the total population netted. Large tench may be unsusceptible to rotenone at the low concentration of 0–5 ppm used, so no check on the accuracy of the population estimation was possible. 相似文献
16.
星斑川鲽雌核发育二倍体、单倍体与普通二倍体及杂交胚胎发育的比较 总被引：1，自引：0，他引：1
通过紫外线(UV)对冷冻的鲈(Lateolabrax japonicus)精子进行灭活,利用冷休克和压力休克方法诱导星斑川鲽(Platichthys stellatus)雌核发育二倍体,同时利用灭活鲈精子制备单倍体胚胎,未灭活鲈精子受精制备杂交胚胎,星斑川鲽精子受精制备正常发育胚胎。对以上几种胚胎发育时序、发育生物学特征进行了观察比较。结果表明,卵裂期单倍体、杂交二倍体和雌核发育二倍体胚胎发育速度与普通二倍体胚胎没有明显差异,从低囊胚期开始各实验组胚胎发育速度均慢于普通二倍体胚胎;杂交胚胎在胚体形成期基本死亡,单倍体胚胎在尾芽期停止发育死亡,均不能正常孵出。雌核发育二倍体与普通二倍体具有相似的发育时序,普通二倍体100 h 10 min孵化出膜,而雌核发育二倍体104 h 50 min孵化出膜。雌核发育胚体畸形率(53.59±0.36)%,孵化率(0.11±0.01)%;普通二倍体胚体畸形率(35.11±6.19)%,孵化率(58.01±5.30)%;与普通二倍体相比,雌核发育二倍体胚体畸形率高,孵化率低,但孵化鱼苗能够正常发育,获得了雌核发育群体。本研究为星斑川鲽雌核发育提供了技术方法,同时为单倍体、杂交和雌核发育胚胎的发育生物学研究提供了细胞生物学证据。 相似文献
17.
Various procedures for artificial insemination in tench, Tinca tinca (L.) were re-examined with evaluation of fecundity of males and females among different tench strains. The objectives of
this study were to enhance fertilization and hatching rates through optimization of the activation solution, the insemination
process, the activation of gametes, and the elimination of eggs stickiness. Sperm for all experiments was collected directly
into immobilization solution of modified Kurokura solution containing 180 mM of NaCl and stored at 2 °C for 2.5–5 h prior
to the experiment. When dechlorinated tap water was used for activation a gamete ratio of 1150 spermatozoa per egg showed
the best significant fertilisation and hatching rates. Optimal ratio between eggs (weight in g) and activation solution (in
cm3) was 1:1. Different concentrations of activation solutions such as NaCl from 0 to 68 mM (0–136 mOsmol kg−1) without buffer statistically decreased fertilization and hatching rates. The activation solution containing 17 mM of NaCl,
10 mM Tris–HCl, pH 8 and 9 significantly increased fertilization and hatching rates compared to dechlorinated tap water of
pH 7 or activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 6 and 7. Adhesiveness of the eggs was successfully
removed by incubation in Alcalase and activity: 3.16 Anson units per cm3. 相似文献
18.
Juan Pablo Alcántar-Vázquez Silvie Dumas Eleonora Puente-Carreón Hugo Skyol Pliego-Cortés & Renato Peña 《Aquaculture Research》2008,39(1):59-63
Conditions for the induction of triploidy with cold shock of fertilized eggs of the spotted sand bass Paralabrax maculatofasciatus (Steindachner) were investigated. Different temperatures (12, 8 and 4 °C), timing of cold shock application (5, 10 and 15 min after fertilization) and duration of the shock (5, 10, 15 and 20 min) were tested. Triploidy was determined using flow cytometry at 12 h after larvae hatched. Triploids were produced only when the cold shock treatment was applied 5 min after fertilization. No significant difference was observed in the percentage of triploidy between temperature and the shock duration. At 8 and 4 °C, 100% triploidy was obtained at different durations of cold shock. Survival was significantly lower at 12 or 4 °C than at 8 °C. No significant difference was observed for shock duration at the temperature of 8 or 12 °C; however, at 4 °C, survival was significantly lower at longer durations. We recommend induction of triploidy by applying cold shock at 8 °C for a duration of 15–20 min starting at 5 min after fertilization, in the spotted sand bass. 相似文献
19.
异源精子诱导条斑星鲽雌核发育 总被引：1，自引：0，他引：1
用冷冻保存的花鲈精子作为异源精子,采用紫外线（UV）对精子进行照射使其遗传物质失活,然后与卵子进行授精,可以刺激条斑星鲽鱼卵进行雌核发育,在受精后一定时间采用冷休克处理“受精”卵,抑制第二极体排出成功获得了条斑星鲽雌核发育二倍体鱼苗。实验表明,花鲈精子只有经过紫外线照射遗传失活后,才能诱导产生条斑星鲽雌核发育二倍体鱼苗。经过大量实验筛选出诱导条斑星鲽雌核发育的最佳条件为花鲈冷冻精子采用80 MJ/cm2 的紫外线照射,然后与条斑星鲽卵子进行授精,在受精后7~9 min,将受精卵放在-1.0~1.5 ℃海水中进行冷休克处理,持续时间为60~90 min,在此条件下,获得的雌核发育二倍体的相对诱导率达40.68%±7.24%。由于不失活精子与条斑星鲽卵形成的杂交胚只能存活到原肠期,而染色体未被成功加倍的胚胎会由于单倍体的致死效应在孵化前后死亡,所以存活的仔鱼全部为条斑星鲽雌核发育二倍体。采用微卫星标记技术对雌核发育鱼苗进行了分析,证明了雌核发育鱼苗为雌核发育二倍体。首次报道了采用异源冷冻精子诱导条斑星鲽鱼卵进行雌核发育的技术方法,为条斑星鲽性别控制和遗传改良提供了技术手段。 相似文献
20.
Heat shock was applied to fertilized eggs of European catfish, Silurus glanis L., for the induction of triploidy. A heat shock of 40. 5°C lasting 1 min and starting 9 min after gamete activation gave the best results with 88.93% of hatched viable fry (92.41% in control group). Yields of hatched viable triploid fry reached 63.14% or 50.60%, when expressed as percentage of the absolute number of viable fry or number of living eggs in eye-bud stage, respectively (P < 0.05). 相似文献