RESUMO
The present work was to study the genetic variability between the major carps Labeo rohita and Cirrhinus mrigala and their hybrids of L. rohita (maleâ) and C. mrigala (femaleâ). Genetic variability was studied by employing RAPD molecular markers. 25 samples of each target species having different sizes with the same age group for the determination of interspecific variation were collected. The morphometric parameters such as body weight, total length, tail length, and lengths of dorsal and anal fins of each individual were recorded and results showed that wet body weight, total length, dorsal fin, anal fin, and tail fin length are positively correlated and then the DNA was extracted using the inorganic salt-based method and conformed by Gel electrophoresis. Twenty-four arbitrary decamer primers were used to get species-specific RAPD analysis Distinct and highly reproducible RAPD profiles with significant genetic variability was detected among species. Only five primers showed amplification. The RAPAD primer OPB-05 produced a total of seven bands out of these 5 monomorphic and 2 polymorphic, so in this case, the percentage polymorphism was 28.57%. The Hybrid show more than a 50% difference from the Labeo rohita. This shows that the Hybrid more resembles C.mrigala. Phylogenetic analysis demonstrated that hybrid (L. rohita â X Cirrhinus mrigala â) is the closest to C. mrigala and the farthest from L. rohita. Overall data are presented concerning the applications of RAPD markers for hybrid identification, genetic diversity assessment, and studying taxonomic relationships at a molecular level.
Assuntos
Carpas , Cyprinidae , Animais , Filogenia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Cyprinidae/genética , Polimorfismo Genético , Variação GenéticaRESUMO
This study examined the artificial breeding and embryonic development of a mountainous fish species Garra cambodgiensis (Tirant, 1883) found in Promlok waterfall in Khaoluang National park, Phromkhiri district, Nakhon Si Thammarat province. The fishes were collected from June 2017- January 2018 and kept in aquaria. Afterward, the brood males and females were selected and injected with buserelin (LHRHa) (10 µg/kg body weight) and domperidone (10 mg/kgbody weight). After the injections, both females and males were kept together in the water at a proportion of 3 females: 1male. The fertilization of eggs started after 4 hours and 30minutes. The fertilized eggs were greenish-grey and semi-buoyant. After fertilization, one blastodisc turned into 2 equalsized blastomeres, and then each cell divided into 4, 8, 16, 32,64 cells respectively. The morula stage went to the blastula stage in about 3hr and 28 min, to gastrula stage in about 5 hr and 11 min, and to the somite stage in about 7 hr and 7 min. The optical vesicles and auditory vesicles developed after approximately 8 hr and 27 min, and 10 hr and 30 min, respectively. After approximately 12 hours and 58 minutes of fertilization, hatching of eggs occurred. Nowadays, the numbers of G. Cambodgienesis are declining rapidly in southern Thailand due to several environmental and anthropogenic reasons. Therefore, it is very important to conserve the populations of G. Cambodgienesis. Artificial breeding could be an effective way to conserve and restore this fish in their natural habitat in southern Thailand.
Assuntos
Masculino , Feminino , Animais , Gravidez , Cyprinidae/crescimento & desenvolvimento , Cyprinidae/embriologia , Cyprinidae/genética , Domperidona/administração & dosagem , Técnicas de Reprodução Assistida/veterináriaRESUMO
This study examined the artificial breeding and embryonic development of a mountainous fish species Garra cambodgiensis (Tirant, 1883) found in Promlok waterfall in Khaoluang National park, Phromkhiri district, Nakhon Si Thammarat province. The fishes were collected from June 2017- January 2018 and kept in aquaria. Afterward, the brood males and females were selected and injected with buserelin (LHRHa) (10 µg/kg body weight) and domperidone (10 mg/kgbody weight). After the injections, both females and males were kept together in the water at a proportion of 3 females: 1male. The fertilization of eggs started after 4 hours and 30minutes. The fertilized eggs were greenish-grey and semi-buoyant. After fertilization, one blastodisc turned into 2 equalsized blastomeres, and then each cell divided into 4, 8, 16, 32,64 cells respectively. The morula stage went to the blastula stage in about 3hr and 28 min, to gastrula stage in about 5 hr and 11 min, and to the somite stage in about 7 hr and 7 min. The optical vesicles and auditory vesicles developed after approximately 8 hr and 27 min, and 10 hr and 30 min, respectively. After approximately 12 hours and 58 minutes of fertilization, hatching of eggs occurred. Nowadays, the numbers of G. Cambodgienesis are declining rapidly in southern Thailand due to several environmental and anthropogenic reasons. Therefore, it is very important to conserve the populations of G. Cambodgienesis. Artificial breeding could be an effective way to conserve and restore this fish in their natural habitat in southern Thailand.(AU)
Assuntos
Animais , Masculino , Feminino , Gravidez , Domperidona/administração & dosagem , Cyprinidae/embriologia , Cyprinidae/crescimento & desenvolvimento , Cyprinidae/genética , Técnicas de Reprodução Assistida/veterináriaRESUMO
Salamanders and lungfishes are the only sarcopterygians (lobe-finned vertebrates) capable of paired appendage regeneration, regardless of the amputation level. Among actinopterygians (ray-finned fishes), regeneration after amputation at the fin endoskeleton has only been demonstrated in polypterid fishes (Cladistia). Whether this ability evolved independently in sarcopterygians and actinopterygians or has a common origin remains unknown. Here we combine fin regeneration assays and comparative RNA-sequencing (RNA-seq) analysis of Polypterus and axolotl blastemas to provide support for a common origin of paired appendage regeneration in Osteichthyes (bony vertebrates). We show that, in addition to polypterids, regeneration after fin endoskeleton amputation occurs in extant representatives of 2 other nonteleost actinopterygians: the American paddlefish (Chondrostei) and the spotted gar (Holostei). Furthermore, we assessed regeneration in 4 teleost species and show that, with the exception of the blue gourami (Anabantidae), 3 species were capable of regenerating fins after endoskeleton amputation: the white convict and the oscar (Cichlidae), and the goldfish (Cyprinidae). Our comparative RNA-seq analysis of regenerating blastemas of axolotl and Polypterus reveals the activation of common genetic pathways and expression profiles, consistent with a shared genetic program of appendage regeneration. Comparison of RNA-seq data from early Polypterus blastema to single-cell RNA-seq data from axolotl limb bud and limb regeneration stages shows that Polypterus and axolotl share a regeneration-specific genetic program. Collectively, our findings support a deep evolutionary origin of paired appendage regeneration in Osteichthyes and provide an evolutionary framework for studies on the genetic basis of appendage regeneration.
Assuntos
Ambystoma mexicanum/genética , Evolução Biológica , Ciclídeos/genética , Cyprinidae/genética , Proteínas de Peixes/genética , Peixes/genética , Regeneração/genética , Ambystoma mexicanum/classificação , Nadadeiras de Animais/fisiologia , Animais , Ciclídeos/classificação , Cyprinidae/classificação , Extremidades/fisiologia , Proteínas de Peixes/classificação , Peixes/classificação , Ontologia Genética , Anotação de Sequência Molecular , Filogenia , TranscriptomaRESUMO
We used next-generation sequencing technology to characterize 19 genomic simple sequence repeat (SSR) markers and 11 expressed sequence tag (EST) SSR markers from Leuciscus leuciscus baicalensis, a small freshwater fish that is widely distributed in Xinjiang, China. Primers were used to test for polymorphisms in three L. leuciscus baicalensis populations in Xinjiang. There were 4-27 (average 11.3) alleles (NA), the expected heterozygosity (HE) was 0.36-0.94 (average 0.75 ± 0.14), the observed heterozygosity (HO) was 0.37-1.00 (average 0.68 ± 0.18), and the polymorphism information content (PIC) was 0.31-0.93 (average 0.71). The averages of HE and PIC for the EST-SSR markers were slightly lower than for the genomic SSR markers. Genetic analysis of the three populations showed similar results for PIC, HE, and NA. Amplifications were performed in nine other species; the top three transferability values were for Rutilus lacustris (80%), Leuciscus idus (76.7%), and Phoxinus ujmonensis (63.3%), with the following average values: PIC (0.56, 4.46, and 0.52); NA (0.40, 3.00, and 0.32); and HO (0.44, 2.74, and 0.22), respectively. L. leuciscus baicalensis is one of the most important commercial fish in Xinjiang, but in recent years, fishery resources have decreased sharply owing to water conservation projects, unreasonable utilization, and invasion by alien species. These novel SSR markers are appropriate for studies involving fingerprinting, gene flow, genetic diversity, population structure, and molecular-assisted breeding, and could contribute to the conservation of L. leuciscus baicalensis.
Assuntos
Cyprinidae/genética , Transferência Genética Horizontal , Repetições de Microssatélites , Polimorfismo Genético , Alelos , Animais , Etiquetas de Sequências Expressas , HeterozigotoRESUMO
To assess the genetic diversity, structure, and population dynamics of Schizopygopsis pylzovi, we examined the changes in mitochondrial DNA sequences (the mtDNA control region and the Cyt b gene; 1835 bp) in 304 individuals from nine populations. The samples were segregated into 112 haplotypes, with high haplotype diversity and low nucleotide diversity. The haplotype diversity was highest in the Minhe (HS) range of Huangshui River and lowest in the Weiyuan (WY) range of Weihe River. Analysis of molecular variance showed that 69.64% of the total genetic variance was contributed by within-the-group variation and 30.36% was contributed by among-the-group variation. Pairwise FST revealed significant divergence between the populations. The FST between the MT and WY was highest, and that between the YZ and YJ was lowest. The neighbor-joining phylogenetic tree demonstrated that all geographic populations were not monophyletic, but overlapped each other, indicating that the duration of geographical isolation was not long enough or the populations had not yet reached significant genetic isolation or differentiation at the monophyletic level. Tajima's D and Fu's Fs were negative and statistically significant, indicating that S. pylzovi had experienced certain population expansion events, which is consistent with the hypothesis that the headwater area of the Yellow River was dramatically affected by the geological and climatic upheaval during the Quaternary ice age. Our analysis indicated that the management units corresponding to the WY population should be managed and conserved first. In situ conservation is first recommended to protect the original habitat from further destruction.
Assuntos
Cyprinidae/genética , Citocromos b/genética , DNA Mitocondrial/genética , Genoma , Filogenia , Animais , China , Conservação dos Recursos Naturais , Cyprinidae/classificação , Variação Genética , Haplótipos , Filogeografia , Dinâmica Populacional , Rios , Análise de Sequência de DNARESUMO
The slender shiner Pseudopungtungia tenuicorpa (Cypriniformes; Cyprinidae; Gobioninae) is an endangered freshwater fish species endemic to Korea. The current strategies for its conservation involve the study of population genetic characters and identification of management units. These strategies require suitable molecular markers to study genetic diversity and genetic structure. Here, we developed nine polymorphic microsatellite markers for P. tenuicorpa for the first time by applying an enrichment method from a size-selected genomic library. The developed microsatellite markers produced a total of 101 alleles (average 11.2). The observed and expected heterozygosities averaged 0.805 and 0.835, respectively. Among the nine identified markers, five markers showed successful amplification across five related Korean Gobioninae species. Thus, the microsatellite markers developed in this study will be useful to establish conservation strategies for both P. tenuicorpa and other related species.
Assuntos
Cyprinidae/genética , Genética Populacional , Repetições de Microssatélites , Alelos , Animais , Conservação dos Recursos Naturais , Cyprinidae/classificação , Espécies em Perigo de Extinção , Genoma , Biblioteca Genômica , Heterozigoto , Reação em Cadeia da Polimerase , República da Coreia , Especificidade da EspécieRESUMO
Twenty-eight polymorphic microsatellite markers were developed from the transcriptome of Ancherythoculter nigrocauda. These loci were used to characterize the genotypes of 48 individuals. The observed number of alleles per locus ranged from 5 to 11, with an average of 7.7. Expected and observed heterozygosities ranged from 0.437 to 0.978 and from 0.373 to 1.000, respectively. Four of these polymorphic microsatellite loci (HWB14, HWB18, HWB24, and HWB30) deviated significantly from the Hardy-Weinberg equilibrium after use of the sequential Bonferroni correction (P < 0.05). Twenty of the 28 loci could be successfully amplified in Culter alburnus. These novel markers will be useful for germplasm resource conservation and management of A. nigrocauda and C. alburnus.
Assuntos
Cyprinidae/genética , Repetições de Microssatélites , Alelos , Animais , Loci Gênicos , Heterozigoto , Polimorfismo Genético , TranscriptomaRESUMO
Skeletal muscle growth is regulated by both positive and negative factors, such as myogenic regulatory factors (MRFs) and myostatin (MSTN), and involves both hyperplasia and hypertrophy. In the present study, morphological changes during muscle development in Megalobrama amblycephala were characterized and gene expression levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) analysis in juvenile [60, 90, 120, and 180 days post-hatching (dph)] and adult fish. Our results show that during muscle development, the frequency of muscle fibers with a diameter <20 µm dramatically decreased in both red and white muscles, with a concomitant increase in the frequency of >30 µm fibers in red muscle and >50 µm fibers in white muscle. At 90-120 dph, the ratio of hyperplastic to hypertrophic areas in red and white muscles increased, but later decreased at 120-180 dph. The effect of hypertrophy was significantly larger than hyperplasia during these phases. qRT-PCR indicated MRF and MSTN (MSTNa and MSTNb) genes had similar expression patterns that peaked at 120 dph, with the exception of MSTNa. This new information on the molecular regulation of muscle growth and rapid growth phases will be of value to the cultivation of M. amblycephala.
Assuntos
Cyprinidae/crescimento & desenvolvimento , Cyprinidae/genética , Regulação da Expressão Gênica no Desenvolvimento , Desenvolvimento Muscular/genética , Músculos/anatomia & histologia , Envelhecimento , Animais , Estatura , Peso Corporal , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Humanos , Hipertrofia , Fibras Musculares Esqueléticas/metabolismoRESUMO
Intermuscular bones, ossified from tendons within the myosepta, occur only in teleost fish. Current understanding of the homology and origins of intermuscular bones in fishes is based mainly on morphological data. To date, there is no published data regarding molecular mechanisms of intermuscular bone formation. In this study, we cloned the gene muscle segment homeobox C (MsxC). MsxC potentially plays a role in intermuscular bone development of Hemibarbus labeo, an important species of cyprinid fish in the Chinese aquaculture industry. Sequence analysis of MsxC revealed motifs characteristic of the homeobox domain family. Whole-mount in situ hybridization showed that MsxC was primarily expressed in the myosepta and brain. MsxC was expressed in the myosepta from 26 to 41 days after hatching (DAH); this coincided with the onset of intermuscular bone ossification, which occurred between 35 and 62 DAH. Evidence for localization of MsxC expression by in situ hybridization correlated with its detection by quantitative real-time PCR. In vertebrates, MsxC plays a role in the regulation of mesenchymal cell differentiation during bone formation. We therefore conclude that MsxC may have a role in epithelium-mesenchyme interactions during intermuscular bone formation in H. labeo.