Complete mitochondrial genome of sterlet (Acipenser ruthenus).

The complete mitochondrial genome of sterlet (Acipenser ruthenus) was determined in this study. The mitogenome is 16,790 bp in length and contains 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions (the control region and the putative origin of the light strand replication) with a typical vertebrate mitochondrial gene arrangement. The overall base composition of the heavy strand is 30.26% for A, 29.00% for C, 16.23% for G and 24.51% for T, with a slight AT bias of 54.77%.

Complete mitochondrial genome of Amur sturgeon (Acipenser schrenckii).

The complete mitochondrial genome of Amur sturgeon (Acipenser schrenckii) was determined in this study. The mitogenome is 16,684 bp in length and contains 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 2 non-coding regions (the control region and the putative origin of the light strand replication) with a typical vertebrate mitochondrial gene arrangement. The overall base composition of the heavy strand is 30.07% for A, 29.36% for C, 16.44% for G and 24.13% for T, with a slight AT bias of 54.20%.

Characterization of a novel nm23 gene and its potential roles in gametogenesis in the prawn Macrobrachium rosenbergii (de Man, 1879) (Crustacea: Decapoda).

Nm23 is a family of genes encoding the nucleoside diphosphate (NDP) kinase, which functions in a wide variety of biological processes, including growth, development, differentiation and tumor metastasis. In this study, a novel nm23 gene, designated as Mrnm23, was identified from the freshwater giant prawn Macrobrachium rosenbergii. The full-length cDNA was 776bp in length, encoding for a protein of 176 amino acids with one typical NDP kinase domain that harbored all the crucial residues for nucleotide binding and enzymatic activity. Like human novel nm23-H1B, the putative protein contained a unique 21-amino-acid NH2-terminal extension as compared to human nm23 (nm23-H1) homologs. Further, 3 extra amino acid residues prolonged the COOH-terminus. The Mrnm23 was ubiquitously expressed in all tissues examined, including androgenic gland, gill, heart, liver, muscle, ovary, and testis. In situ hybridization to gonad sections indicated that the Mrnm23 mRNA was localized in the cytoplasm of cup-base of differentiating spermatids, in the spike of the umbrella-shaped spermatozoa and in the cytoplasm of the early previtellogenic oocytes, suggesting that the Mrnm23 has potential roles in spermiogenesis and early differentiation of oocyte.

[Microsatellite markers for parentage identification in Jian Carp (Cyprinus carpio var. Jian)].

Using 16 microsatellite loci we estimated the parentage of 647 progeny in 10 Jian Carp full-sib families. Cervus 3.0 analysis showed that mean PIC value of 16 microsatellites, mean number of allele, and mean expected heterozygosity were 0.7025, 6.63, and 0.7405, respectively. The combined probability of exclusion was 0.99922456 when both parents were unknown and the combined probability of exclusion was 0.99999557 when only one of the parental genotype was known, with the confidence level of 95%. Further simulations based on allele frequencies suggested that to achieve the requirements of paternity test usually took 8 to 12 microsatellite loci when both parents were unknown and 5 to 8 microsatellite loci when one parent was known. Out of 647 progenies, 94.6% were assigned to their parental pairs without the information of both parents in parentage analysis, which were lower than the theoretical assignment rates predicted by the Cervus simulations. This could be explained by the relationship between the candidate parents or existence of null and by typing errors. The identification of 9 families was useful for linkage analysis of Jian Carp and QTL location, also for marker assisted selection for economical traits.

[DNA barcoding and species and subspecies classification within genus Carassius].

The classification of Carassius has not been well established due to its great variability and wide distribution. Usually, Carassius is identified as three species: C. carassius, C. cuvieri and C. auratus, the latter including several subspecies, such as goldfish. Out of these subspecies, C. auratus gibelio have recently been thought of as a valid species of Carassius. In this study we collected the 5'end 651 bp segments of the mitochondrial cytochrome c oxidase I (COI) gene from 128 specimens, including C. carassius, C. cuvieri, C. auratus auratus, C. auratus gibelio and C. auratus langsdorfii. All three species of Carassius (C. carassius, C. cuvieri, C. auratus) were found to be valid, meanwhile genetic differentiation between the Eurasian C. auratus and Japanese C. auratus has reached a high level. However, several haplotypes were shared between C. auratus auratus and C. auratus gibelio. Consequently, C. auratus gibelio should be regarded as a subspecies of C. auratus rather than a valid species. Moreover, because both diploids and triploids exist in C. auratus auratus and C. auratus gibelio, ploidy level should not be used as criteria for the classification of species or subspecies in Carassius.

[Quantitative trait locus analysis of standard length, body depth and body thickness in mirror carp (Cyprinus carpio L.)].

Based on a full-sib family, the genetic linkage map was constructed with 246 microsatellite and 306 SNP markers, which was used to detect the QTLs for standard length (SL), body depth (H), body thickness (BT), and the ratio of standard length and body depth (SLH) in mirror carp by GridQTL software. The results indicated that a total of 14 related QTLs distributed on the 7 linkage groups were obtained. Seven QTLs were related to standard length, of which the linkage groups of LG6, LG17, LG21, LG23, and LG35 were at 5% significant level, and linkage group LG1 and LG28 were at 1% significant level, which explained 6.6%-12.6% of the phenotypic variance. Three QTLs were identified for body depth on the linkage groups of LG17, LG23 and LG28 (P amp; 0.01), accounting for 11.6%, 12.7%, and 15.6% of the phenotypic variance, respectively. Two QTLs were associated with body thickness on the linkage of LG23 and LG28 (P amp; 0.05), which explained 8.6% and 7.2% of the phenotypic variation, respectively. Two QTLs were responsible for the ratio of standard length and body depth on the linkage of LG21 and LG35 (P amp; 0.05), both of which explained 8.2% of the phenotypic variance. The results provide a useful reference for further candidate gene research and molecular marker assisted selection in mirror carp.

Rates and patterns of microsatellite mutations in common carp (Cyprinus carpio L.).

During genotyping 150 microsatellites in a F1 family of common carp, six mutations were found at five microsatellite loci. The overall mutation rate of common carp microsatellites was 2.53 X 10(-4) per locus per generation. At five loci, mutations increased the length of alleles by at least one repeat unit, suggesting mutations at microsatellite loci in common carp do not follow strict stepwise mutation model. The data on mutation rates and patterns can facilitate population genetics studies, and provide useful parameters for estimating a long-term effective population size of common carp.

[Mapping QTLs related to head length, eye diameter and eye cross of common carp (Cyprinus carpio L.)].

A common carp gynogenetic line including 44 individuals derived from the cross Barbless carp x Hebao-cold tolerance red carp was used to construct a linkage map using 445 markers (265 AFLP markers, 127 SSR markers, 37 EST-SSR markers and 16 RAPD markers). Quantitative traits loci (QTLs) associated with head length, eye diameter, and eye cross were identified by composite interval mapping of the software WinQTLCart2.5. Five QTLs were identified for head length on the linkage group of LG2, LG3, LG40, and LG4, which explained 12.39% to 34.29% of the total variation of the head length. All of their additive effects were negative. Two QTLs were associated with eye diameter on the linkage groups of LG39 (qED-39-1) and LG40 (qED-40-1), which explained 9.77% and 5.62% of the total variation of the eye diameter, respectively. The additive effect of qED-39-1 was positive and that of qED-40-1 was negative. Two QTLs were responsible for eye cross on the linkage group of LG28 and LG20, explaining 8.88% and 8.29% of the total variation of the eye cross, respectively. The additive effect of qEC-28-1 was negative and that of qEC-20-1 was positive.

[Studies on quantitative trait loci related to activity of lactate dehydrogenase in common carp (Cyprinus carpio)].

The reciprocal intergeneric hybrids between common wheat and Roegneria kamoji were successfully obtained by means of embryo culture. Morphology, chromosome pairing behavior at meiosis, fertility, and resistance to scab of the hybrid F1 and BC1 were studied. The results showed that the morphology of the reciprocal intergeneric hybrids F1 between R. kamoji and T. aestivum cv. Chinese Spring were intermediate type between the two parental species. The chromosome configuration at metaphase I (MI) of pollen mother cell (PMC) in reciprocal F1 was 40.33I + 0.78II + 0.03III and 40.40I + 0.79II, respectively. All of the F1 plants showed complete male sterility, and the seeds of BC1 were obtained by backcrossing with Chinese Spring pollen. The somatic chromosome numbers in BC1 plants of (R. kamoji x Chinese Spring) F1 x Chinese Spring ranged from 55 to 63. Many univalents were observed at MI of PMC, which resulted in the sterility of BC1 plants. Similarly, the chromosome numbers in BC1 plants of (Chinese Spring xR. kamoji) F1 x Chinese Spring also ranged from 55 to 63; however, many bivalents at MI of PMC and fertile pollen were observed resulting in partial fruitfulness in some BC1 plants by self-crossing. A plant (2n=63) with 42 wheat chromosomes and 21 R. kamoji chromosomes was obtained from R. kamoji x Chinese Spring cross, which had a chromosome configuration at MI of 26.40I + 18.30II. Because many univalents existed, this plant showed complete male sterility, and BC1 plants were obtained by back-crossing with Chinese Spring as the pollen parent. The chromosome numbers of BC1 ranged from 40 to 59, which contained less alien chromosomes. Although the morphology of the spike in BC1 plants was similar to that of Chinese Spring, these BC1 plants were still sterile. All F1 and most of the BC1 plants showed high resistance to Fusarium graminearum, which indicated that the resistance to scab from R. kmoji can be transferred into wheat.Microsatellite markers were used to make marker regression analysis on activity of lactate dehydrogenase based on double pseudo-testcross strategy using Windows Map Manager2.0 software. The parents that came from the cross between progenies of Hebao-cold tolerance red carp and Barbless carp and F2 progenies were used as segregating populations. For maker regression, a total of 12 markers associated with activity of lactate dehydrogenase were significant at P<0.05 and HLJE222 was significant at P<0.01. The variance explained by these loci, ranged from 4.00% to 10.00%. Locus HLJE222 was closely linked to the gene related to activity of lactate dehydrogenase of common carp. For further identification, EST-SSR markers were used to screen the protein and nucleotide database using bioinformatics tools in order to find the homologies. High sequence similarities of HLJE222 marker were observed with the nucleotide sequence of DAZ associated protein 1mRNA of zebrafish(94%), and protein sequence of DAZ associated protein 1(97%). DAZ protein is one of the short chain dehydrogenases, which is an important enzyme in the process of glucose metabolism in the organisms. This family contains a wide variety of dehydrogenases. This indicates that locus HLJE222 was closely linked to the gene associated with activity of lactate dehydrogenase of common carp.

[Correlation analysis of microsatellite DNA markers with body weight, length and height of common carp (Cyprinus carpio L.)].

Forty-seven microsatellite markers were selected to analyze the genomic DNA of 92 progenies derived from the recombinant inbred lines (RIL) of common carp, which came from the cross between Barbless carp and Hebao-cold tolerance red carp. The results showed that a total of 162 different alleles were found, and the number of alleles in each locus was 2 to 6. The DNA fragment length was 100 bp to 444 bp, and the number of mean valid alleles was 1.3069 to 4.2288. The value of heterozygosity was 0.2361 to 0.7677, and the mean polymorphism information content (PIC) was 0.5368. A GLM procedure was used to analyze the effects of these 47 microsatellites on body weight, length and height. Results uncovered HLJ695, HLJ716, HLJ739, HLJ759, HLJ774 and K16 had a significant impact on body weight, length and height, and HLJ776 had a significant impact on height. In addition, the genotypes of these correlative loci were determined.

[Mutation rate and pattern of microsatellites in gynogenetic silver crucian carp (Carassius autatus gibeblio)].

The natural gynogenetic triploid silver crucian carp (Carassius autatus gibeblio Bloch) provides a good system for studying evolutional genetics of the unisexual and polyploidy vertebrate. Microsatellites are abundant across genomes and show high levels of polymorphism and mutational rate, so they have been widely used for studying evolutional biology. In this study, the mutation rate and pattern at 33 microsatellite loci of silver crucian carp were investigated. As a result, it was found that the only one of 22 offspring had 18 mutant alleles at 15 microsatellite loci. The overall mutation rate of the 33 loci was 1.16x10(-2)/locus/generation (95% confidence interval 6.87x10(-3) and 1.83x10(-2)). The mutation rate in the gynogenetic triploid silver crucian carp was obviously higher than other fish, which was closely related to the transitional phase of parthenogenesis and gamogenesis in the natural gynogenetic fish. The repeat numbers had more than 10 times at 13 loci of the mutant alleles, and there was no obviously different in the mutant rate between the 11 compound microsatellite loci (1.31x10(-2) )and the 21 perfect microsatellite loci(1.00x10(-2)) (P = 0.67). The mutant rate had affinity with repeat numbers instead of repeat types and GC content in flanking sequences of microsatellite. The mutation pattern of silver crucian carp was very complexional, as well as some loci did not follow the Stepwise Mutation Model.

[Analysis of genetic structure of mirror carp population and correlation of microsatellite markers and economic traits].

The genetic structure of one breeding population of mirror carp(Cyprinus carpio L.) was analysed using thirty-five polymorphic microsatellite markers. The effective number of alleles (Ae), observed heterozygosity (Ho), expected heterozygosity (He) and polymorphism information content(PIC) were all determined. The Hardy-Weinberg equilibrium was checked by chi-square test. For each locus, 2-7 alleles were detected, with a total of 118 alleles for 35 loci . The value of Ae, Ho, He and PIC showed that the genetic variation of the population was not high .The average effective number of alleles and the mean polymorphism information content were 2.16 and 0.42, respectively, and the observed and expected heterozygosity were 0.431 and 0.4736 , respectively.The probability value of chi-square test showed that more than half of the thirty-five loci have significantly (Plt;0.01) deviated from Hardy-Weinberg equilibrium. The correlation of genotype of each locus and individual phenotype data was analysed , and 4 loci seemed associated with body weight and body size . When compared with the result of QTL mapping of common carp, HLJ319 locus was consistent with the mapping result approximately, which was significantly correlated with body size . Several significantly deviated loci were also analysed, and the possible cause of the kind of deviation was discussed .

Development of 10 tri- and tetranucleotide microsatellite loci for population studies in the common carp (Cyprinus carpio L.).

Ten tri- and tetranucleotide microsatellite DNA markers were isolated and characterized from common carp (Cyprinus carpio L.) to estimate genetic potential. These markers were tested in the samples from two closely related carp populations (Cyprinus carpio var. xingguonensis and Cyprinus carpio var. wananensis). The number of the alleles ranged from three to nine, and observed and expected hererozygosities varied from 0.207 to 1.000 and from 0.499 to 0.900 in each population, respectively. No evidence for linkage disequilibrium was found, indicating that these markers will be useful for population studies.

[Genetic potential analysis of Germany mirror carp (Cyprinus carpio L.) using microsatellite markers].

Using 30 microsatellite markers and combining quantity characters such as body weight, body size and body width, we evaluated the genetic potential of 3 Germany mirror carp populations. Number of effective alleles (Ae), observed (Ho) and expected (He) heterozygosity values and polymorphic information contents (PIC) were all calculated. 287 alleles and 559 genotypes were detected. The DNA fragment length was 109-400 bp. The Hardy-Weinberg Equilibrium was checked and phenomenon of some disequilibrium were studied according to the test of c2. The result showed that the level of genetic variability was moderate, but genetic potential of Shuanglai population was much lower than that Huanxin and Songpu breeding populations. Polymorphic Information Contents (PIC) of the 3 populations of Germany mirror carp were between 0.08787 and 0.5377, both of highly and moderately polymorphic markers were 13. The number of effective alleles (Ae) were between 1.1014 and 6.4665. The observed (Ho) and expected (He) heterozygosity values were 0.0968-0.9892 and 0.0926-0.8554 respectively. The linkage correlation was analyzed using data of body weight, body size and body width and 30 loci. The result shown that there existed 2 loci, HLJ319 and HLJ693 associated with body size. The HLJ693 locus significantly correlated with body weight character. The HLJ677 locus linked with body width. And then the result were verified in Recombinant Inbred Lines (RIL) of common carp. It shown that the HLJ319 locus was significantly linked with body size, the same as the result of common carp QTL location.

[Mapping and genetic effect analysis of quantitative trait loci related to body size in common carp (Cyprinus carpio L.)].

The common carp recombinant inbred lines (RIL) derived from the cross Barbless carp x Hebao-cold tolerance red carp were used as experimental materials in this study. Based on the linkage map constructed with simple sequence repeat (SSR) markers using this RIL population, marker regression and complexity interval mapping were analyzed by Windows Map Manager2.0 software. A P-value of 0.01 was the threshold value of single-marker. A linkage group-wide permutation test (1 000 replicates) determined the significance of the maximum LOD value over the various intervals analyzed for each linkage group. The results are as follows: 1) For maker regression, a total of 7 makers related body size, were at 1% significant level, of which 3 were at 0.1% significant level. The variance explained by these loci, ranged 14.00% to 27.00%. Loci HLJ534, HLJ319, and HLJ370 at 0.1% significant level closely linked to the gene related to body size of common carp; 2) six significant QTLs related to body size were at the 5% linkage group-wide level on these linkage groups, and two of them were at 1% level. The variances explained by these QTLs ranged from 11.33% to 23.12% and their additive effects were not identical. HLJ190-HLJ497 and HLJ479-HLJ483 were major QTLs associated to body size of common carp.