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1.
Gene–centromere mapping using half‐tetrad analysis is a powerful tool for understanding chromosomal behaviour and determining the position of centromeres in relation to genes or markers in fish. In this study, eight gynogenetic diploid families induced by inhibiting of the second meiotic division were genotyped at 66 microsatellite loci for microsatellite–centromere (M‐C) mapping in bighead carp ( Aristichthys nobilis). The absence of paternal alleles verified the success of gynogenetic development in all gynogenetic families. All loci were consistent with Mendelian segregation in control families. The estimated recombination frequency (y) ranged from 0.057 to 0.875 with an average of 0.477 ± 0.222. Seventeen loci (25.76%) showed high M‐C recombination frequencies of over 0.667. M‐C distances ranged from 2.85 to 43.75 cM under the assumption of complete interference. Thus, these loci are distributed from the centromeres to the telomeres of their respective chromosomes, while mainly in the intermediate region. Information on centromere mapping could serve as a starting point to consolidate the genetic linkage groups in bighead carp.  相似文献   

2.
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   

3.
To develop an effective system for parentage analysis in gynogenetic and clonal progeny of Nile tilapia, Oreochromis niloticus L., polymorphic microsatellite loci and amplified fragment length polymorphisms (AFLPs) were investigated in several gynogenetic families and clonal lines. Six microsatellite loci were screened in two meiotic gynogenetic families to look for loci with high gene–centromere recombination rates, which can be used to discriminate meiotic from mitotic gynogenetics. Microsatellite loci UNH189 and UNH211 showed 96.7% and 92.0% heterozygosity, respectively, in these families, while other loci showed lower recombination frequencies. Scoring both UNH189 and UNH211 would give a very low probability of an individual meiotic gynogenetic being homozygous for both loci. Multiplex polymerase chain reaction of microsatellite loci was used to verify parentage in four families of mitotic gynogenetics and five fully inbred clonal lines. The genotype of each clonal line should serve as a unique identifier. Twelve AFLP primers were also investigated and 26 diagnostic AFLP bands were identified to follow inheritance in mitotic gynogenetic individuals. Amplified fragment length polymorphisms were found to be effective for this purpose but microsatellites were more appropriate since they are co‐dominant, while AFLPs are dominant markers. A multiplex of the microsatellite loci used in this study would be useful for general parental assignment as well as for the analysis of the products of chromosome set manipulations.  相似文献   

4.
A fast and cost‐effective protocol to develop candidate microsatellite markers from sea perch, Lateolabrax japonicus, was described here. Ten suites of codominant bands that contained seven microsatellites were discovered in this marine fish, in which no microsatellite development was reported previously. All the seven microsatellites were found to be polymorphic among tested 20 individuals of sea perch. Five of the above seven loci conformed to Hardy–Weinberg equilibrium (HWE) as determined by using the Markov‐Chain method implemented, the other two loci significantly deviated from HWE, both of them showed a large heterozygote excess (HE). Out of 21 possible pair‐wise comparisons among the seven loci applied to sea perch, none showed significant linkage disequilibrium (LD) (P > 0.008). Five additional fish species assessed for cross‐species amplification revealed that some of the loci appear to be applicable in close‐related species.  相似文献   

5.
The turbot (Scophthalmus maximus) is a flatfish species of great commercial value for aquaculture. In this study we describe the isolation and characterization of 30 novel highly polymorphic microsatellite markers in this species obtained from a genomic library enriched for seven short tandem repeated motifs. Much higher polymorphism (mean number of alleles: 13.37; mean expected heterozygosity: 0.869) and potential for parentage assignment than previously reported for microsatellites in turbot were found after the analysis of 24 wild individuals. Most loci conformed to Hardy–Weinberg expectations, excluding Sma‐USC20 and Sma‐USC28, which showed a high heterozygote deficit probably due to the presence of null alleles. No significant genotypic disequilibrium was observed between any pair of loci, suggesting no close linkage between them. These loci are potentially useful for kinship and population analysis in turbot.  相似文献   

6.
微卫星标记在长牡蛎三倍体鉴定中的应用   总被引：1，自引：1，他引：0  
姜群  李琪  于红  孔令锋 《水产学报》2014,38(12):1970-1975
为开发一种简单有效的倍性检测方法对长牡蛎三倍体诱导结果进行准确评估,本实验采用细胞松弛素B抑制第二极体释放诱导产生长牡蛎三倍体,选用7个微卫星位点扩增基因组DNA,通过亲子代基因分型进行倍性检测,检测结果采用流式细胞仪加以验证,以评估微卫星标记倍性检测的准确性。另外,本研究探讨了准确鉴定倍性所需的微卫星标记数量与微卫星—着丝粒重组率(y)之间的关系,以期为其他物种采用分子标记进行倍性检测提供理论依据。结果显示,细胞松弛素B诱导产生的115个长牡蛎子代经7个微卫星位点鉴定得到40个三倍体,与流式细胞仪检测结果一致,准确率达到100%,7个微卫星位点的(1-y)的乘积为0.005。随机挑选6个位点,也可鉴定出所有三倍体,(1-y)的乘积为0.005~0.042。研究表明,本研究中开发的微卫星标记可以简单高效地鉴定长牡蛎三倍体,微卫星位点的(1-y)的乘积小于0.005,倍性检测的准确率可达100%,采用微卫星标记进行倍性检测对于加速长牡蛎三倍体育种进程具有十分重要的意义。  相似文献   

7.
Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.  相似文献   

8.
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9.
This study evaluated the properties of nine Scylla paramamosain microsatellite loci screened by us previously for inclusion in a parentage assignment marker suite. These nine highly polymorphic markers (mean He=0.847 and PIC=0.830) were determined as being suitable for parentage assignment. Simulations based on allele frequency data from 15 known maternal families (165 individuals) demonstrated that at least four loci were required to assign >95% of offspring to maternal parents with 95% confidence. In actual parentage assignments, all progenies were assigned to the maternal parents with six or more loci, which was similar to the simulation predictions. Our results suggest that this set of microsatellites provide a powerful and efficient tool for identifying pedigree information for selective breeding programmes of S. paramamosain.  相似文献   

10.
In order to develop a microsatellite typing system for Lepeophtheirus salmonis (Krøyer), a DNA preparation method for individual sea lice suitable for analysis by polymerase chain reaction (PCR) was designed, and the DNA sequences of 50 L. salmonis microsatellite elements were determined. The microsatellites were composed of 60% perfect, 25% imperfect, and 15% compound repeats. Based on the flanking DNA sequences, four microsatellite‐PCR assays were optimized and used in a pilot study to analyse L. salmonis samples collected in Ireland, Norway and Scotland. Two of the microsatellite‐PCR assays targeted polymorphic loci amplifying seven and 10 alleles respectively. The results showed that microsatellite‐PCR typing could detect genetic variation both within and between the L. salmonis groups, and also was capable of amplifying group‐specific alleles.  相似文献   

11.
Parentage analysis in aquaculture determines genealogical relationships between broodstock and progeny when the parents are unknown. Thus, parentage analysis is a useful tool to establish pedigree reports in molecular‐assisted selection programs. Here, we evaluated 10 heterologous microsatellite markers for parentage assignment in abalone hybrids produced from 43 abalone broodstocks of red abalone (Haliotis rufescens) and Japanese abalone (H. discus hannai). The allele frequencies, exclusion probabilities and broodstock contributions were calculated using CERVUS, PAPA and GERUD software. The polymorphic information content (PIC) values showed that most of the microsatellite loci were highly informative (>0.7) and more than 90% of parentage assignment was possible with a minimum of 5–6 microsatellite markers. Parentage assignment for hybrid and pure‐red progeny showed a better performance than pure‐Japanese progeny. This result could be due to the high level of allele loss in the parental genotypes. In addition, results indicated that only two sires contributed over 80% and 90% of red and hybrid progenies, respectively. This study gives a new molecular tool to support marker‐assisted selection in abalone hybrids produced in Chile.  相似文献   

12.
以乌江流域中华倒刺鲃(Spinibarbus sinensis)为材料,构建了中华倒刺鲃微卫星富集文库。以乌江彭水电站所采集的32个个体的基因组DNA为模板,进行PCR扩增,结果筛选出8个多态性微卫星位点,其等位基因数4~10,多态信息含量0.6072~0.8515;观测杂合度0.5625~1.0000,期望杂合度0.6791~0.8795,Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡偏离指数-0.2009~0.3805;卡方检验结果表明,3个位点不同程度地偏离了Hardy中华倒刺鲃微卫星标记筛选及特征分析Weinberg平衡(P<0.05);各位点两两之间不存在显著的连锁不平衡现象。  相似文献   

13.
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   

14.
Aeromonas hydrophila is a pathogen infecting farmed hybrid catfish, Clarias macrocephalus (Günther, 1864) × Clarias gariepinus (Burchell, 1822) which incurs substantial economic losses in Thailand. The study aimed at a genetic tracking of A. hydrophila infection and the in vitro assessment of the efficacy of antibiotics against its virulent strains. Five clinical strains from catfishes and Nile tilapia were employed. They were 3‐passage re‐isolated through healthy hybrid catfish and the cytolytic enterotoxin gene (AHCYTOEN) of individuals was traced. Each of the re‐isolates at a dose of ~6.67 × 105 CFU/g was intraperitoneally injected into ~15 g‐healthy hybrid catfish and their pathogenicity were observed for 7 days. It was found that AHCYTOEN was carried over whereas typical signs of motile aeromonas septicaemia were found in the specimens. The bacterial strains of Nile tilapia origin did not induce mortality but those of catfish origins (80%–100% rate of mortality). The strains were susceptible to the tetracycline antibiotics, and oxytetracycline produced MIC50 and MBC as low as 0.007–0.031 μg/ml and 1–8 μg/ml respectively. As oxytetracycline specifically inhibited pathogenic A. hydrophila in vitro, it is recommended that an appropriate dosage regimen of the drug should be established.  相似文献   

15.
剑尾鱼微卫星DNA的筛选   总被引：19，自引：4，他引：19       下载免费PDF全文
以剑尾鱼(Xiphophorus helleri)为材料,经MboⅠ限制性内切酶消化基因组DNA后,选取500～2 000 bp的片段连接到经BamHⅠ酶切的pUC18载体上,转化大肠杆菌DH5α构建部分基因组文库.采用设计合成的(AC)7、(GT)7重复序列为引物,PCR筛选部分基因组文库,对其中9个重组阳性克隆进行测序,结果共获得24个微卫星序列,其中Perfect(完美型)13个,占54.2%;Imperfect(非完美型)3个,占12.5%;Compound(混合型)8个,占33.3%.表明(AC/GT)n在剑尾鱼的基因组DNA中含量非常丰富.同时,根据其中3个克隆微卫星的侧翼序列设计引物,PCR扩增剑尾鱼基因组DNA,结果均扩增到目的片段.而且,这3对引物扩增出来的微卫星片段在非选育的剑尾鱼中显示出多态性,而在近交系19代则表现为单态,为剑尾鱼的实验动物化遗传研究提供了理论依据.  相似文献   

16.
Parentage analysis is of vital importance for hatchery production and breeding programmes. Two multiplex PCR protocols including seven tropical spiny lobster (Panulirus homarus) microsatellites (Pho‐G06, Pho‐G25, Pho‐G53, Pho‐G62, Pho‐G74, Pho‐G89 and Pho‐G100) were introduced for parentage assignment. All loci were polymorphic with allele sizes from 113 to 337 base pairs (bp), observed alleles (k) from two to seven and polymorphic information content (PIC) of 0.25 to 0.73. Twenty‐four stage‐3 phyllosoma larvae (39 days posthatching) were collected for paternity exclusion study using selected microsatellites. Exclusion‐based parentage analysis unambiguously assigned 83% of fry (20 of 24) to a single female parent. Of ten putative female parents, five have contributed to the 20 allocated offspring, with one being true parent of 11. Four others were assigned to two or more potential female parents, probably due to genotyping error or the presence of null allele. The exclusion power (EP) for the seven loci varied between 13% and 54% with known genotypes of one parent (P1) and 19% and 71% for given the genotypes of both parents (P2). The theoretical combined parentage exclusion (cEP) power for all seven microsatellites was P1 = 95% and P2 = 99%. The power of discrimination for each locus varied between 0.18 and 0.86. This report presents the first study to utilize microsatellite markers for successful parentage assignment of P. homarus. The selected microsatellites provide a practical tool for parentage analysis in hatchery production of juveniles as well as in future commercial breeding programme of tropical spiny lobsters.  相似文献   

17.
This study was designed to determine the effect of ovarian fluid pH of stripped unfertilized channel catfish, Ictalurus punctatus, eggs on fertilization and hatch rate of channel catfish ♀ x blue catfish, Ictalurus furcatus♂, hybrid catfish eggs. A significant correlation was established between ovarian fluid pH of stripped channel catfish eggs and hybrid embryo hatch rate (R2 = 0.75, P = 0.01), suggesting ovarian fluid pH of stripped catfish eggs prior to fertilization can be predictive of the hatching success of hybrid catfish embryos. These data were used to categorize pH of stripped eggs: pH <7.0 as “low pH eggs,” pH 7.0–7.4 as “medium pH eggs,” or pH >7.4 as “high pH eggs” quality eggs. The range in percent hatch rate for these pH categories was <15%, 15–30%, and >30%, respectively. Higher calcium concentrations during incubation do not appear to improve hatching success of “low pH eggs.” The predictive model presented herein describes a quick method for evaluating stripped catfish eggs for hybrid fry production in catfish hatcheries.  相似文献   

18.
This study describes the genetic structure of five naturalized populations of brown trout in southern Chile using allozyme and microsatellite markers to establish levels of intra‐ and interpopulation genetic variability and divergence. Fourteen enzymatic systems were used comprising 20 loci and three microsatellite loci specific to brown trout. The genetic variability values (allozymes, P=20–35%, average=27%, HO=0.118–0.160, average=0.141; microsatellites, P=33.3–100%, average=66.66%, HO=0.202–0.274, average=0.229) are similar to values described in other naturalized populations of brown trout present in Chile, but higher than those observed in European populations of this species. Values of total genetic diversity (HT) (allozymes=0.1216 and microsatellites=0.3504) and relative genetic divergence (GST) (allozymes=9.5% and microsatellites=15%) were also similar to the results obtained in previous studies of Chilean populations of brown trout. These values, when compared with those obtained in Europe, proved to be similar for HT but lower for GST. The low interpopulational genetic differentiation was in accordance with the small genetic distance observed between the populations analysed (D Nei=0.004–0.025). On the other hand, the high frequency of one of the two alternative alleles of the phylogeographic marker locus LDH‐5* in the populations analysed (LDH‐5*90>0.84) would indicate a European origin, in particular Atlantic as opposed to Mediterranean, for the brown trout introduced into Chile. The high levels of genetic variability suggest a mixed origin for the naturalized brown trout in Chile, which could have originated either before or during the introduction process. Nevertheless, the low level of genetic differentiation between populations could reflect the short lapse of time in evolutionary terms, during which populations introduced into Chile have been exposed to different evolutionary forces, and which has not been sufficiently long to produce greater genetic differentiation between populations.  相似文献   

19.
云斑尖塘鳢微卫星分子标记的筛选与检测   总被引：1，自引：0，他引：1  
用磁珠富集法分离云斑尖塘鳢的微卫星序列,由所获得的1032个克隆中筛选出146个阳性克隆,经测序81个含有微卫星序列,52个为完美型,27个为非完美型,2个为复合型,其中43个微卫星序列重复次数在10以上。所获得的云斑尖塘鳢微卫星序列中除探针中使用的CA重复单元和GA重复单元外,还有TAC等其他类型的重复单元。设计合成38对微卫星引物,其中29对引物可稳定扩增出条带,使用这些引物对云斑尖塘鳢48个个体进行检测显示:观测杂合度平均值为0.63,期望杂合度平均值为0.43。29对引物中1对引物表现为单态,7对表现为高度多态,14对表现为中度多态,7对表现为低度多态,多态性较为丰富,说明本研究开发的绝大部分微卫星分子标记较为理想。  相似文献   

20.
The demographics (e.g. hatch time, growth and mortality) and habitat (macrohabitat and mesohabitat) use of age‐0 blue catfish Ictalurus furcatus (Lesueur) were evaluated in the un‐impounded reach of the middle Mississippi River during 2003–2010. Each year, island complexes and adjacent main river habitats were trawled, and 7,373 age‐0 blue catfish (10–144 mm total length) were collected. Blue catfish hatch timing varied slightly across years and generally began in June and ended in August. Age‐0 blue catfish growth rates and instantaneous mortality rates differed among years. Catch rates of age‐0 blue catfish were highest in side channels and around islands, whereas main channel habitats were infrequently occupied. Regarding mesohabitat use, age‐0 blue catfish frequently occupied low velocity areas (i.e. 0.3–0.4 m/s) of shallow depth (i.e. 1–4 m) and sand substratum. This study has provided insight into age‐0 blue catfish population dynamics and has highlighted the importance of specific habitat characteristics in large rivers that may aid in recruitment of this species.  相似文献   

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