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1.
There is an increasing requirement for traceability of aquaculture products, both for consumer protection and for food safety. There are high error rates in the conventional traceability systems depending on physical labels. Genetic traceability technique depending on DNA-based tracking system can overcome this problem. Genealogy information is essential for genetic traceability, and microsatellite DNA marker is a good choice for pedigree analysis. As increasing genotyping throughput of microsatellites, microsatellite multiplex PCR has become a fast and cost-effective technique. As a commercially important cultured aquatic species, Pacific oyster Crassostrea gigas has the highest global production. The objective of this study was to develop microsatellite multiplex PCR panels with dye-labeled universal primer for pedigree analysis in C. gigas, and these multiplex PCRs were validated using 12 full-sib families with known pedigrees. Here we developed six informative multiplex PCRs using 18 genomic microsatellites in C. gigas. Each multiplex panel contained a single universal primer M13(?21) used as a tail on each locus-specific forward primer and a single universal primer M13(?21) labeled with fluorophores. The polymorphisms of the markers were moderate, with an average of 10.3 alleles per locus and average polymorphic information content of 0.740. The observed heterozygosity per locus ranged from 0.492 to 0.822. Cervus simulations revealed that the six panels would still be of great value when massive families were analysed. Pedigree analysis of real offspring demonstrated that 100% of the offspring were unambiguously allocated to their parents when two multiplex PCRs were used. The six sets of multiplex PCRs can be an important tool for tracing cultured individuals, population genetic analysis, and selective breeding program in C. gigas. 相似文献
2.
Iwagaki oyster,Crassostrea nippona,widely distributes along the seashore of Eastern Asia,and has been considered to be a potential breeding species due to its delicious taste and high commercial value.In order to study its genetic background and population structure,we developed 46 novel polymorphic microsatellite markers using next-generation sequencing technique and characterized them in 30 individuals.The number of alleles ranged from 3 to 22,while the observed and expected heterozygosities varied from 0.133 to 1.000 and 0.455 to 0.949,respectively.Fifteen microsatellite markers were selected and grouped into five highly informative multiplex PCRs for C.nippona.We evaluated and validated these multiplex PCRs in a cultured population including 173 candidate parents and 486 offspring.In actual parentage analysis,80%of the offspring were correctly assigned to their parental pairs using three multiplex PCRs.Furthermore,the success rate of parentage assignment reached 96%when the other two multiplex PCRs were added.These 46 microsatellite loci with high variability and the five multiplex PCRs described here provide a powerful tool for pedigree reconstruction,resource conservation and selective breeding program of C.nippona. 相似文献
3.
《中国海洋大学学报(英文版)》2016,(2)
Scapharca broughtonii is a commercially important and over-exploited species.In order to investigate its genetic diversity and population structure,43 novel polymorphic microsatellites were isolated and characterized.The number of alleles per locus ranged from 3 to 22 with an average of 6.93,and the observed and expected heterozygosities varied between 0.233 and 1.000,and 0.250 and 0.953,with an average of 0.614 and 0.707,respectively.Three highly informative multiplex PCRs were developed from nine of those microsatellites for S.broughtonii.We evaluated and validated these multiplex PCRs in 8 full-sib families.The average polymorphism information content(PIC) was 0.539.The frequency of null alleles was estimated as 3.13% of all the alleles segregation based on a within-family analysis of Mendelian segregation patterns.Parentage analysis of real offspring demonstrated that 100% of all offspring were unambiguously allocated to a pair of parents based on 3 multiplex sets.Those 43 microsatellite loci with high variability will be helpful for the analysis of population genetics and conservation of wild stock of S.broughtonii.The 3 sets of multiplex PCRs could be an important tool of pedigree reconstruction,population genetic analysis and brood stock management. 相似文献
4.
Scapharca broughtonii is a commercially important and over-exploited species. In order to investigate its genetic diversity and population structure, 43 novel polymorphic microsatellites were isolated and characterized. The number of alleles per locus ranged from 3 to 22 with an average of 6.93, and the observed and expected heterozygosities varied between 0.233 and 1.000, and 0.250 and 0.953, with an average of 0.614 and 0.707, respectively. Three highly informative multiplex PCRs were developed from nine of those microsatellites for S. broughtonii. We evaluated and validated these multiplex PCRs in 8 full-sib families. The average polymorphism information content (PIC) was 0.539. The frequency of null alleles was estimated as 3.13% of all the alleles segregation based on a within-family analysis of Mendelian segregation patterns. Parentage analysis of real offspring demonstrated that 100% of all offspring were unambiguously allocated to a pair of parents based on 3 multiplex sets. Those 43 microsatellite loci with high variability will be helpful for the analysis of population genetics and conservation of wild stock of S. broughtonii. The 3 sets of multiplex PCRs could be an important tool of pedigree reconstruction, population genetic analysis and brood stock management. 相似文献
5.
Five full-sib families of the Pacific oyster(Crassostrea gigas) larvae were used to study the mode of inheritance at eight microsatellite loci,and the feasibility of these markers for kinship estimate was also examined.All eight microsatellite loci were compatible with Mendelian inheritance.Neither evidence of sex-linked barriers to transmission nor evidence of major barriers to fertilization between gametes from the parents was shown.Three of the eight loci showed the presence of null alleles in four families,demonstrating the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies.Although the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance,offspring from five full-sib families were unambiguously discriminated in the neighbor-joining dendrogram.This result indicates that the microsatellite markers may be capable of discriminating between related and unrelated oyster larvae in the absence of pedigree information,and is applicable to the investigation of the effective number of parents contributing to the hatchery population of the Pacific oyster. 相似文献
6.
The inheritance mode of seven microsatellite markers was investigated in Patinopecten yessoensis larvae from four controlled crosses, and the feasibility of using these markers for kinship estimation was also examined.
All the seven microsatellite loci were compatible with Mendelian inheritance. Neither sex-linked barriers to transmission
nor major barriers to fertilization between gametes from the parents were evident. Two of the seven loci showed the presence
of null alleles in two families, suggesting the need to conduct comprehensive species-specific inheritance studies for microsatellite
loci used in population genetic studies. However, even if the null allele heterozygotes were considered as homozygotes in
the calculation of genetic distance, offspring from four families were all unambiguously discriminated in the neighbor-joining
dendrogram. This result indicates that the microsatellite markers used may be capable of discriminating between related and
unrelated scallop larvae in the absence of pedigree information, and of investigating the effective number of parents contributing
to the hatchery population of the Japanese scallop. 相似文献
7.
Five full-sib families of the Pacific oyster （Crassostrea gigas） larvae were used to study the mode of inheritance at eight microsatellite loci, and the feasibility of these markers for kinship estimate was also examined. All eight microsatellite loci were compatible with Mendelian inheritance. Neither evidence of sex-linked barriers to transmission nor evidence of major barriers to fertilization between gametes from the parents was shown. Three of the eight loci showed the presence of null alleles in four families, demonstrating the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies. Although the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance, offspring from five full-sib families were unambiguously discriminated in the neighbor-joining dendrogram. This result indicates that the microsatellite markers may be capable of discriminating between related and unrelated oyster larvae in the absence of pedigree information, and is applicable to the investigation of the effective number of parents contributing to the hatchery population of the Pacific oyster. 相似文献
8.
《中国海洋大学学报(英文版)》2017,(1)
Glycogen is important not only for the energy supplementary of oysters, but also for human consumption. High glycogen content can improve the stress survival of oyster. A key enzyme in glycogenesis is glycogen synthase that is encoded by glycogen synthase gene GYS. In this study, the relationship between single nucleotide polymorphisms(SNPs) in coding regions of Crassostrea gigas GYS(Cg-GYS) and individual glycogen content was investigated with 321 individuals from five full-sib families. Single-strand conformation polymorphism(SSCP) procedure was combined with sequencing to confirm individual SNP genotypes of Cg-GYS. Least-square analysis of variance was performed to assess the relationship of variation in glycogen content of C. gigas with single SNP genotype and SNP haplotype. As a consequence, six SNPs were found in coding regions to be significantly associated with glycogen content(P 0.01), from which we constructed four main haplotypes due to linkage disequilibrium. Furthermore, the most effective haplotype H2(GAGGAT) had extremely significant relationship with high glycogen content(P 0.0001). These findings revealed the potential influence of Cg-GYS polymorphism on the glycogen content and provided molecular biological information for the selective breeding of good quality traits of C. gigas. 相似文献
9.
The inheritance mode of seven microsatellite markers was investigated in Patinopecten yessoensis larvae from four con-trolled crosses, and the feasibility of using these markers for kinship estimation was also examined. All the seven microsatellite loci were compatible with Mendelian inheritance. Neither sex-linked barriers to transmission nor major barriers to fertilization between gametes from the parents were evident. Two of the seven loci showed the presence of null alleles in two families, suggesting the need to conduct comprehensive species-specific inheritance studies for microsatellite loci used in population genetic studies. However, even if the null allele heterozygotes were considered as homozygotes in the calculation of genetic distance, offspring from four fami-lies were all unambiguously discriminated in the neighbor-joining dendrogram. This result indicates that the microsatellite markers used may be capable of discriminating between related and unrelated scallop larvae in the absence of pedigree information, and of investigating the effective number of parents contributing to the hatchery population of the Japanese scallop. 相似文献
10.
Breeding practice for Undaria pinnatifida (Harvey) Suringar requires the screening of a large number of offspring from gametophyte crossings to obtain an elite variety for large-scale cultivation. To better understand the genetic relationships of different gametophyte cultures isolated from different sources, 20 microsatellite loci were screened and 53 gametophyte clone cultures analyzed for U. pinnatifida isolated from wild sporophytes in Vladivostok, Russia and from cultivated sporophytes from Dalian and Qingdao, China. One locus was abandoned because of poor amplification. At the sex-linked locus of Up-AC-2A8, 3 alleles were detected in 25 female gametophyte clones, with sizes ranging from 307 to 316 bp. At other loci, 3 to 7 alleles were detected with an average of 4.5 alleles per locus. The average number of alleles at each locus was 1.3 and 3.7 for Russian and Chinese gametophyte clones, respectively. The average gene diversity for Russian, Chinese, and for the combined total of gametophyte clones was 0.1, 0.4, and 0.5, respectively. Russian gametophyte clones had unique alleles at 7 out of the 19 loci. In cluster analysis, Russian and Chinese gametophyte clones were separated into two different groups according to genetic distance. Overall, high genetic diversity was detected in gametophyte clones isolated from the two countries. These gametophyte cultures were believed to be appropriate parental materials for conducting breeding programs in the future. 相似文献
11.
Genetic diversity and differentiation of the oyster Crassostreaplicatula populations from China's coast were studied basedon seven microsatellite loci.All loci showed high polymorphism for all five C.plicatula populations,with an average number of alleleper locus of 19.3-27.9 and an average expected heterozygosity of 0.889-0.952.Significant departures from Hardy-Weinberg equilib-rium and deficits of heterozygotes were observed over most populations at each locus,which were fully explained by null alleles.Microsatellite analysis revealed significant subdivision in the C.plicatula populations.According to the neighbor-joining tree con-structed on the basis of the D,A distance,the five populations fell into three regional groups,showing a relatively homogeneous geneticstructure in geographically close populations.Assignation tests correctly assigned high percentages of individuals to their originalpopulations and groups,and also confirmed the existence of genetic differentiation among C.plicatula populations.The results ob-tained in this study will facilitate the formulation of appropriate fisheries management programs,stock identification and conservationof biodiversity for the species. 相似文献
12.
《中国海洋大学学报(英文版)》2021,(4)
Shell color is regarded as an economic trait in many breeding programs of bivalve mollusks, but the relationship between shell color and production traits remains controversial. In our breeding program of Crassostrea gigas, solid white, black, and orange shell lines were obtained, and second-generation(F2) and testcross families with segregating shell colors were constructed by crossing these three lines. These segregating families provided ideal samples for investigating the relationship between shell color and production traits in C. gigas. The growth and survival of 7-month-old oysters with different shell colors sampled within the same families were compared in seven F2 families and 13 testcrosses. In addition, the growth and survival of oysters from the three shell color lines were compared at 4 and 16 months of age. The growth and survival rates of the orange shell line were significantly lower than those of the white and black shell lines. However, no significant difference in growth between oysters with different shell colors was observed within segregating families, except the testcrosses produced by crossing orange-and white-shelled grandparents, and no significant difference in survival was observed in any family. Overall, no significant correlation was observed between shell color and production traits in C. gigas. These results suggest that shell color cannot be used as a marker to guide the selection of growth and survival traits. Thus shell color and production traits should be selected independently in oyster breeding programs. 相似文献
13.
The annual life cycle of the brown seaweed Undaria pinnatifi da(Harvey)Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage.In 2011,an unusual zoospore-derived monoecious gametophyte isolate(designated as line 10-5-3)of U.pinnatifi da was observed.To understand this phenomenon,a comprehensive screening of eighty-two previously identifi ed male gametophyte cultures,isolated from three randomly selected cultivars(lines 10,7,and 5)was performed.Thirty-six of the isolates developed both antheridia and oogonia on the same fi lamentous fragment in a standard gametogenesis test(SGT: 18°C,60 μmol photons/(m 2·s)).Selfi ng of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring.However,crossing resulted in a much higher fertilization rate(89.7%).The hybrid and selfed sporophytic offspring were grown to maturity in fl ow tanks at an ambient temperature of 10–18°C over a period of 69 days.Active zoospores were released from both types of mature sporophylls.The majority of these developed into male gametophytes,while 15%–20% developed into the observed monoecious structures on the same fi lament.Using PCR amplifi cation it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfi ng and crossing lacked the femalelinked microsatellite sequence(a part of the locus Up-AC-2A8,GenBank accession No.AY738602.1),indicating their male nature.U.pinnatifi da is an invasive species in some regions and the implications of the above fi ndings for this species in nature are briefl y discussed. 相似文献
14.
15.
Genetic diversity and differentiation of the oyster Crassostrea plicatula populations from China’s coast were studied based on seven microsatellite loci. All loci showed high polymorphism for all
five C. plicatula populations, with an average number of allele per locus of 19.3–27.9 and an average expected heterozygosity of 0.889–0.952.
Significant departures from Hardy-Weinberg equilibrium and deficits of heterozygotes were observed over most populations at
each locus, which were fully explained by null alleles. Microsatellite analysis revealed significant subdivision in the C. plicatula populations. According to the neighbor-joining tree constructed on the basis of the D
A distance, the five populations fell into three regional groups, showing a relatively homogeneous genetic structure in geographically
close populations. Assignation tests correctly assigned high percentages of individuals to their original populations and
groups, and also confirmed the existence of genetic differentiation among C. plicatula populations. The results obtained in this study will facilitate the formulation of appropriate fisheries management programs,
stock identification and conservation of biodiversity for the species. 相似文献
16.
Inheritance of 15 microsatellites in the Pacific oyster Crassostrea gigas： segregation and null allele identification for linkage analysis 总被引：2，自引：0，他引：2
Microsatellites were screened in a backcross family of the Pacific oyster, Crassostrea gigas. Fifteen microsatellite loci were distinguishable and polymorphic with 6 types of allele-combinations. Null alleles were detected in 46.7% of loci, accounting for 11.7% of the total alleles. Four loci did not segregate in Mendelian Ratios. Three linkage groups were identified among 7 of the 15 segregating loci. Fluorescence-based automated capillary electrophoresis (ABI 310 Genetic Analyzer) that used to detect the microsatellite loci, has been proved a fast, precise, and reliable method in microsatellite genotyping. 相似文献
17.
18.
军曹鱼全人工繁殖群体遗传特征的SSR分析 总被引：2，自引：1，他引：1
利用8个微卫星DNA位点分析南海海域5个军曹鱼全人工繁育群体(HN1、HN2、ZJ、FJ和LS)子代的遗传多样性特征和群体间遗传分化。结果显示,军曹鱼养殖群体与天然群体的遗传结构特征基本一致:1)平均有效等位基因数为3.910±0.440,观测杂合度为0.595±0.049,分子方差分析(AMOVA)表明,个体内分化占主导(46%),军曹鱼养殖群体整体遗传多样性较高;2)群体间基因流明显(N_m=2.5959,F_(st)=0.0878),整体分化程度较低。各养殖群体表现出不同于天然大群体的特征:1)绝大部分位点均明显偏离哈温平衡,杂合子缺失或过剩现象普遍存在;2)聚类和群体分配分析等表明HN2与另四个养殖群体(HN1、ZJ、FJ和LS)分化明显。 相似文献
19.
《中国海洋大学学报(英文版)》2016,(3)
Gene-centromere mapping is an essential prerequisite for understanding the composition and structure of genomes. Half-tetrad analysis is a powerful tool for mapping genes and understanding chromosomal behavior during meiosis. The Japanese scallop(Patinopecten yessoensis), a cold-tolerant species inhabiting the northwestern Pacific coast, is a commercially important marine bivalve in Asian countries. In this study, inheritance of 32 informative microsatellite loci was examined in 70-h D-shaped larvae of three induced meiogynogenetic diploid families of P. yessoensis for centromere mapping using half-tetrad analysis. The ratio of gynogenetic diploids was proven to be 100%, 100% and 96% in the three families, respectively. Inheritance analysis in the control crosses showed that 51 of the 53 genotypic ratios observed were in accordance with Mendelian expectations at the 5% level after Bonferroni correction. Seven of the 32 microsatellite loci showed the existence of null alleles in control crosses. The second division segregation frequency(y) of the microsatellite loci ranged from 0.07 to 0.85 with a mean of 0.38, suggesting the existence of positive interference after a single chiasma formation in some chromosomes in the scallop. Microsatellite-centromere distances ranged from 4 c M to 42 c M under the assumption of complete interference. Information on the positions of centromeres in relation to the microsatellite loci will represent a contribution towards the assembly of genetic maps in the commercially important scallop species. 相似文献
20.
The sea cucumber Apostichopus japonicus is a commercially and ecologically important species in China. A total of 3056 potential unigenes were generated after assembling 7597 A. japonicus expressed sequence tags (ESTs) downloaded from Gen-Bank. Two hundred and fifty microsatellite-containing ESTs (8.18%) and 299 simple sequence repeats (SSRs) were detected. The average density of SSRs was 1 per 7.403 kb of EST after redundancy elimination. Di-nucleotide repeat motifs appeared to be the most abundant type with a percentage of 69.90%. Of the 126 primer pairs designed, 90 amplified the expected products and 43 showed polymorphism in 30 individuals tested. The number of alleles per locus ranged from 2 to 26 with an average of 7.0 alleles, and the observed and expected heterozygosities varied from 0.067 to 1.000 and from 0.066 to 0.959, respectively. These new EST-derived microsatellite markers would provide sufficient polymorphism for population genetic studies and genome mapping of this sea cucumber species. 相似文献