Zhaobin Song

Assessing genetic diversity of wild populations of Prenantȁ9s schizothoracin, Schizothorax prenanti, using AFLP markers

Prenantȁ9s schizothoracin, Schizothorax prenanti, an endemic fish to China, has undergone a dramatic decline in numbers due to human impacts. We studied its genetic diversity in three tributaries of the Yangtze River: the Qingyi River, which has many hydropower dams, and the Dadu River and Muli River where many hydropower dams are being proposed. Using amplified fragment length polymorphism (AFLP), 621 loci were amplified with seven AFLP primer combinations in 45 individuals. The loci were highly polymorphic and heterozygous (87% polymorphism, 30% heterozygosity). The genetic distances within populations were large. The analysis of molecular variance demonstrated that most variation occurred within populations. The estimated fixation index (Φst) value averaged over all polymorphic loci across the three rivers was 0.0837, indicating a moderate genetic differentiation. The differentiations between populations were significant, and population structure was strong. The results suggested that China had wild populations of Prenantȁ9s schizothoracin with considerable genetic diversity in the Muli, Dadu and Qingyi rivers. The proposals to dam these rivers should take into account the importance of conserving their genetic quality.

Population genetic diversity of Prenant’s schizothoracin, Schizothorax prenanti , inferred from the mitochondrial DNA control region

Wild populations of Prenant’s schizothoracin, Schizothorax prenanti, were collected from the Qingyi, Dadu, and Muli rivers in upper reaches of the Yangtze River. Genetic variation and population structure were examined based on analysis of the mitochondrial DNA control region. Nucleotide sequence analysis defined 30 haplotypes in 41 individuals. The nucleotide and haplotype diversities were very high, and genetic distances between the Dadu and Qingyi populations were much smaller than between either of them and the Muli population. The analysis of molecular variance demonstrated that most variation occurred within samples, and the differentiation between Muli and Dadu or Qingyi was significant. Estimates of gene flow indicated reproductive isolation between Muli and the other populations, and this divergence might be related to geographical location. The results are mostly consistent with the findings from AFLP analysis and also suggested considerable genetic diversity of the populations in the Muli, Dadu, and Qingyi rivers.

Complete mitochondrial genome of the stone loach, Triplophysa stoliczkae (Teleostei: Cypriniformes: Balitoridae).

The complete mitochondrial genome of the stone loach Triplophysa stoliczkae is 16,571 bp in size, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and a noncoding control region. As in other vertebrates, most mitochondrial genes are encoded on the heavy strand, except for nd6 and eight tRNA genes which are encoded on the light strand. The overall base composition of the heavy strand of the T. stoliczkae mitochondrial genome is A: 28.1%, T: 29.0%, C: 25.0%, and G: 17.9%. The alignment of the Triplophysa species control regions exhibited high genetic variability and rich A+T content.

Complete mitochondrial genome of a new vole Proedromys liangshanensis (Rodentia: Cricetidae) and phylogenetic analysis with related species: are there implications for the validity of the genus Proedromys?

Aim. The complete mitochondrial genome sequence of a newly discovered vole, Proedromys liangshanensis (Rodentia: Cricetidae: Arvicolinae), was determined. Results. The mitogenome of P. liangshanensis is 16,296 bp in length. As with most other mammals, it contains the same gene order and an identical number of genes or regions, including 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one putative control region. The A+T content of the control region is 58.2%, the lowest proportion detected so far in Myomorpha. To confirm the phylogenetic position of P. liangshanensis, we carried out phylogenetic analyses based on complete mitochondrial genomic data using Bayesian, maximum parsimony, and maximum likelihood methods. Conclusion. All results revealed that P. liangshanenis is sister to Microtus. Although the results do not bear light on the validity of the genus Proedromys, based on the morphological characters, we suggest that Proedromys is an independent genus of equal rank to the genus Microtus.

The complete mitochondrial genome of the taimen, Hucho taimen, and its unusual features in the control region

The whole mitochondrial genome of Hucho taimen was firstly sequenced and characterized. The genome is 16,833 bp in length and contains 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a noncoding control region. Twelve protein-coding genes on the heavy strand showed that the content of A+T was higher than that of G+C, whereas the nd6 protein-coding gene on the light strand displayed an opposite pattern. We described the secondary structure of the origin of light strand (oriL) replication and found that the conserved 5′-GCCGG-3′ sequence motif is variable in H. taimen and some other salmonids. We conclude that the control region is variable in length and represents the high A+T content, compared with other mitochondrial control regions available in Salmonidae and other non-salmonids. Additionally, another interesting feature of H. taimen mitogenome is that a T-type mononucleotide microsatellite and an 82 bp tandem repeat were identified in the control region.

Transcriptome analysis of the plateau fish (Triplophysa dalaica): Implications for adaptation to hypoxia in fishes

Triplophysa dalaica, endemic species of Qinghai-Tibetan Plateau, is informative for understanding the genetic basis of adaptation to hypoxic conditions of high altitude habitats. Here, a comprehensive gene repertoire for this plateau fish was generated using the Illumina deep paired-end high-throughput sequencing technology. De novo assembly yielded 145, 256 unigenes with an average length of 1632 bp. Blast searches against GenBank non-redundant database annotated 74,594 (51.4%) unigenes encoding for 30,047 gene descriptions in T. dalaica. Functional annotation and classification of assembled sequences were performed using Gene Ontology (GO), clusters of euKaryotic Orthologous Groups (KOG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. After comparison with other fish transcriptomes, including silver carp (Hypophthalmichthys molitrix) and mud loach (Misgurnus anguillicaudatus), 2621 high-quality orthologous gene alignments were constructed among these species. 61 (2.3%) of the genes were identified as having undergone positive selection in the T. dalaica lineage. Within the positively selected genes, 13 genes were involved in hypoxia response, of which 11 were listed in HypoxiaDB. Furthermore, duplicated hif-α (hif-1αA/B and hif-2αA/B), EGLN1 and PPARA candidate genes involved in adaptation to hypoxia were identified in T. dalaica transcriptome. Branch-site model in PAML validated that hif-1αB and hif-2αA genes have undergone positive selection in T.dalaica. Finally, 37,501 simple sequence repeats (SSRs) and 19,497 high-quality single nucleotide polymorphisms (SNPs) were identified in T. dalaica. The identified SSR and SNP markers will facilitate the genetic structure, population geography and ecological studies of Triplophysa fishes.

Complete mitochondrial genomes of two lenoks, Brachymystax lenok and Brachymystax lenok tsinlingensis

The circular mitochondrial genomes of Brachymystax lenok and B. lenok tsinlingensis are 16,832 and 16,669 bp in length, respectively. The mitogenomes of two lenoks shared common features with those of other teleosts in terms of gene arrangement, base composition, and transfer RNA structures. The two genomes have an overall nucleotide sequence identity of 98.3%. Through the comparisons between the two lenoks, we found their gene arrangement, composition, and sizes are the same, and the A+T content is identical. As with other freshwater salmonids, a T-type mononucleotide microsatellite and various tandem repeats were identified in the control regions of the lenoks. The low pairwise distance (2.1%) inferred from 12 mitochondrial protein-coding genes on heavy strand showed close proximity of B. lenok and B. lenok tsinlingensis.

Isolation and characterization of nine polymorphic microsatellite loci in the rock carp, Procypris rabaudi (Tchang).

Nine highly polymorphic microsatellite loci were isolated from AC‐ and GATA‐repeat microsatellite enrichment DNA libraries in the rock carp, Procypris rabaudi (Tchang). The number of alleles for these loci ranged from eight to 18 in tested individuals. Polymorphism information content ranged from 0.712 to 0.908 with an average of 0.837. Average observed and expected heterozygosities were 0.719 and 0.870, respectively. These molecular markers will be useful for the assessment of genetic diversity and analysis of population structure in wild rock carp.

Taxonomic implications from phylogenetic relationships of subspecies of Schizopygopsis malacanthus (Pisces: Cyprinidae) based on sequence analysis of cytochrome b and mitochondrial DNA control region

Phylogenetic relationships of three subspecies of Schizopygopsis malacanthus, S. m. malacanthus, S. m. chengi, and S. m. baoxingensis, were investigated based on the mitochondrial DNA control region and the cytochrome b gene. Phylogenetic analysis indicated that the three subspecies did not cluster as one monophyletic group; S. m. malacanthus clustered into one clade, while S. m. chengi and S. m. baoxingensis clustered into another. Genetic distances between S. m. malacanthus and the other two subspecies were either very close to or larger than those between S. m. malacanthus and some other species of Schizopygopsis. There was very small genetic distance between S. m. chengi and S. m. baoxingensis. The results suggested S. m. chengi should be split from S. malacanthus into a separate species, Schizopygopsis chengi (Fang); S. m. baoxingensis should be regarded as a subspecies, S. c. baoxingensis (Fu, Ding et Ye), of Schizopygopsis chengi.

High intra-population genetic variability and inter-population differentiation in a plateau specialized fish, Triplophysa orientalis

Triplophysa orientalis (Herzenstein) is one of the Nemacheilinae (Cypriniformes: Balitoridae) fish species distributed in the Tibetan Plateau area. In order to understand the impact of plateau uplift on population history and the isolation effect of plateau lakes on T. orientalis, we examined its genetic structure and phylogenetic relationships. A total of 98 individuals from five wild populations, three from plateau lakes and two from branch rivers in upper reaches of the Yangtze River, in the eastern peripheral of the Tibetan Plateau were sampled. An 848 base pair fragment from the mitochondrial DNA (mtDNA) control region was sequenced for analyses. Overall, very high intra-population genetic variability was found in all populations except for one lake population (Rannicuo); nucleotide diversity ranged from 0.0025 to 0.0159 and haplotype diversity ranged from 0.641 to 0.879. Furthermore, the genetic distance between river populations (0.0326) was much higher than that among lake populations (Rannicuo and Barencuo 0.0035, Bannicuo and Yibicuo 0.0038, Rannicuo and Yibicuo 0.0049). Additionally, the analysis of molecular variance demonstrated that most of the observed genetic variability occurred among populations, accompanied with significant Fst values except for that between the Yibicuo and Barencuo populations. This evidence suggested a strong population structure of the species and a lack of inter-population connection. Lastly, the rate of migration indicated there were large historic gene flows among lake populations. Demographic analysis also indicated there were bottlenecks or expansions in three lake populations, suggesting a potential isolation effect of plateau lakes on population differentiation. Molecular dating of intra-specific divergence showed the plateau uplift has shaped the genetic structure of T. orientalis.