Jinzhong Fu

Genetic variability among endangered Chinese giant salamanders, Andrias davidianus.

The endangered Chinese giant salamander (Andrias davidianus) is endemic to mainland China. Genetic divergence among six populations of the species was investigated by means of isozyme electrophoresis and mitochondrial DNA (mtDNA) sequences. Forty allozyme loci were resolved for all populations; the amount of genetic divergence among populations was comparable to that in other amphibians. mtDNA sequences showed a similar level of divergence. The population from Huangshan is distinct from other populations, indicating the existence of localized divergence. Both allozyme and mtDNA data failed to associate the populations into a pattern corresponding to the three Chinese river systems, which may be the consequence of human relocation. Conservation policies should emphasize the protection of localized populations and cessation of human‐facilitated introductions. Future studies should focus on investigating the divergence among localized populations from isolated mountain regions, particularly using more fine‐grained techniques such as microsatellite DNA.

Speciation in the Rana chensinensis species complex and its relationship to the uplift of the Qinghai–Tibetan Plateau

Speciation remains a fundamental issue in biology. Herein, we report an investigation into speciation in the Rana chensinensis species complex using DNA sequence data from one mitochondrial and five nuclear genes. A phylogenetic analysis of the data revealed four major clades in the complex, and each of them was found to likely represent a species, including one cryptic species. Ecological niche models were generated from 19 climatic variables for three of the four major clades, which were represented by widespread sampling, including R. chensinensis, Rana kukunoris and the potential cryptic species. Each clade is associated with a unique ecological unit, and this indicates that ecological divergence probably drove speciation. Ecological divergence is likely related to the late Cenozoic orogenesis of the Qinghai–Tibetan Plateau. In addition, gene flow between species was detected but only in peripheral portions of the ranges of the four major clades, thus likely had little influence on the speciation processes. Discordances between mitochondrial and nuclear genes were also found; the nominal species, R. chensinensis, contains multiple maternal clades, suggesting potential mitochondrial introgression between R. chensinensis and R. kukunoris.

Toward understanding the genetic basis of adaptation to high-elevation life in poikilothermic species: A comparative transcriptomic analysis of two ranid frogs, Rana chensinensis and R. kukunoris

BackgroundUnderstanding how organisms adapt to high-elevation environments at a genome scale provides novel insights into the process of adaptive evolution. Previous studies have mainly focused on endothermic organisms, while poikilothermic species may have evolved different mechanisms to cope with high-elevation environments. In this context, we sequenced transcriptomes of a pair of closely related anuran species, Rana chensinensis and R. kukunoris, which inhabit respective low- and high-elevation habitats. By comparing the two transcriptomes, we identified candidate genes that may be involved in high-elevation adaption in poikilothermic species.ResultsOver 66 million sequence reads from each transcriptome were generated. A total of 41,858 and 39,293 transcripts for each species were obtained by de novo assembly. By comparing the orthologous transcripts, we identified 125 protein-coding genes that have likely experienced strong positive selection (Ka/Ks>1). In addition, 335 genes that may bear a signature of positive selection (1≥Ka/Ks>0.5) were also recognized. By considering their functions, fourteen candidate genes were determined to be likely involved in high-elevation adaptation, including two CYP genes, USP-1, and several others.ConclusionsWe identified a set of candidate genes that may have promoted adaptation of R. kukunoris to its high-elevation environment. These include several genes that have previously been associated with oxygen binding, response to UV radiation, and repair of free radical injury. Detailed molecular, physiological, and phenotypic studies are required to better understand the roles of these genes in improving the performance of R. kukunoris in a high-elevation environment. We have evidence for both convergent and divergent evolution between endothermic and poikilothemic species, but additional research across a wide range of organisms will be necessary to comprehend the complexity of high-elevation adaptation.

Frequent mitochondrial gene introgression among high elevation Tibetan megophryid frogs revealed by conflicting gene genealogies

Historical mitochondrial introgression causes differences between a species’ mitochondrial gene genealogy and its nuclear gene genealogy, making tree‐based species delineation ambiguous. Using sequence data from one mitochondrial gene (cytochrome b) and three nuclear genes (introns), we examined the evolutionary history of four high elevation Tibetan megophryid frog species, Scutiger boulengeri, Scutiger glandulatus, Scutiger mammatus and Scutiger tuberculatus. The three nuclear genes shared a similar history but the mitochondrial gene tree suggested a drastically different evolutionary scenario. The conflicts between them were explained by multiple episodes of mitochondrial introgression events via historical interspecific hybridization. ‘Foreign’ mitochondrial genomes might have been fixed in populations and extended through a large portion of the species’ distribution. Some hybridization events were probably as old as 10 Myr, while others were recent. An F1 hybrid was also identified. Historical hybridization events among the four species appeared to be persistent and were not restricted to the period of Pleistocene glaciation, as in several other well‐studied cases. Furthermore, hybridization involved several species and occurred in multiple directions, and there was no indication of one mitochondrial genome being superior to others. In addition, incomplete lineage sorting resulting from budding speciation may have also explained some discrepancies between the mitochondrial DNA and nuclear gene trees. Combining all evidences, the former ‘Scutiger mammatus’ appeared to be two species, including a new species. With the availability of a wide range of highly variable nuclear gene markers, we recommend using a combination of mitochondrial gene and multiple nuclear genes to reveal a complete species history.

Divergence of the Cytochrome b Gene in the Lacerta raddei Complex and Its Parthenogenetic Daughter Species: Evidence for Recent Multiple Origins

Abstract Questions concerning the origin of parthenogenesis in Caucasian Rock Lizards and genetic divergence among bisexual lizards of the Lacerta raddei complex were examined using sequences from the mitochondrial cytochrome b gene. The maternal parent of the parthenogenetic L. uzzelli, L. sapphirina, and L. bendimahiensis was confirmed to be L. raddei. Although substantial variation was revealed among bisexual populations of L. raddei and L. nairensis, very low or no variation was found among the parthenogenetic species. A phylogenetic tree including 11 populations of L. raddei and L. nairensis, as well as 10 populations of its five daughter parthenogens, was constructed. Because of paraphyletic relationships, L. nairensis is considered conspecific with L. raddei. Evaluation of the parthenogenetic species suggests that separate hybridization events between L. raddei and L. valentini might have occurred at least twice. One resulted in L. sapphirina and L. bendimahiensis, and the other one (or more) resulted in L. unisexualis and L. uzzelli. The females involved were distantly related, Lacerta unisexualis and L. uzzelli likely had separate origins, but the females involved were closely related.

An unexpected recent ancestor of unisexual Ambystoma.

Previous research has shown that members of the unisexual hybrid complex of the genus Ambystoma possess a mitochondrial genome that is unrelated to their nuclear parental species, but the origin of this mitochondrion has remained unclear. We used a 744‐bp fragment of the mitochondrial gene cytochrome b within a comparative phylogenetic framework to infer the maternal ancestor of this unisexual lineage. By examining a broader range of species than has previously been compared, we were able to uncover a recent maternal ancestor to this complex. Unexpectedly, Ambystoma barbouri, a species whose nuclear DNA has not been identified in the unisexuals, was found to be the recent maternal ancestor of the individuals examined through the discovery of a shared mtDNA haplotype between the unisexuals and A. barbouri. Based on a combination of sequence data and glacial patterning, we estimate that the unisexual lineage probably originated less than 25 000 years ago. In addition, all unisexuals examined showed extremely similar mtDNA sequences and the resultant phylogeny was consistent with a single origin for this lineage. These results confirm previous suggestions that the unisexual Ambystoma complex was formed from a hybridization event in which the nuclear DNA of the original maternal species was subsequently lost.

Does life history shape sexual size dimorphism in anurans? A comparative analysis

BackgroundThe evolution of sexual size dimorphism (SSD) is likely constrained by life history. Using phylogenetic comparative methods, we examined correlations between SSD among anurans and their life history traits, including egg size, clutch size, mating combat, and parental care behaviour. We used sexual dimorphism index (SDI = Body-sizefemale /Body-sizemale –1) as the measurement for SSD. Body size, life history and phylogenetic data were collected from published literature. Data were analysed at two levels: all anuran species and within individual families.ResultsFemale-biased SSD is the predominant form in anurans. SSD decreases along with the body size increase, following the prediction of Rensch’s rule, but the magnitude of decrease is very small. More importantly, female body size is positively correlated with both fecundity related traits, egg size and clutch size, and SDI is also positively correlated with clutch size, suggesting fecundity advantage may have driven the evolution of female body size and consequently leads to the evolution of female-biased SSD. Furthermore, the presence of parental care, male parental care in particular, is negatively correlated with SDI, indicating that species with parental care tend to have a smaller SDI. A negative correlation between clutch size and parental care further suggests that parental care likely reduces the fecundity selection pressure on female body size. On the other hand, there is a general lack of significant correlation between SDI and the presence of male combat behaviour, which is surprising and contradictory to previous studies.ConclusionsWe find clear evidence to support the ‘fecundity advantage hypothesis’ and the ‘parental care hypothesis’ in shaping SSD in anurans. Nevertheless, the relationships of both parental care and combat behaviour to the evolution of SSD are complex in anurans and the extreme diversity of life history traits may have masked some potential interesting relationships. Our study represents the most comprehensive study of SSD in anurans to date.

A phylogeographic analysis of populations of the wood turtle (Glyptemys insculpta) throughout its range.

We investigated the phylogeography of wood turtles, Glyptemys insculpta, in North America using 750 bp of the mitochondrial control region from 117 individuals sampled at 29 localities across the species’ range. A total of 21 haplotypes were identified and little genetic variation was found. The highest pairwise difference was 2%. From nested clade analysis (NCA), one main postglacial dispersal route was inferred along the east coast, with subsequent westward dispersal. NCA further revealed some patterns of restricted gene flow/dispersal. We propose that wood turtles experienced a combined effect of bottleneck during the Pleistocene as well as of selective sweep, which produced the low level of variation observed. Following the Pleistocene, wood turtles would have undergone a rapid northward expansion from a common southern refugium as glaciers retreated. These findings shed light on where to direct conservation priorities, on conservation strategies needed, and on the potential effects of interpopulation transfers for this vulnerable species.

How many species are in the genus Batrachuperus? A phylogeographical analysis of the stream salamanders (family Hynobiidae) from southwestern China

Phylogeographical analysis of DNA sequence data has been routinely used to test species boundaries using the monophyly criterion; however, a complementary criterion, reproductive isolation, is often ignored. We used a combination of phylogenetic and population genetic approaches to determine species boundaries among stream salamanders in the genus Batrachuperus. First, cytochrome b sequence data from 174 Batrachuperus individuals, sampled from 78 populations, were used to reconstruct historical relationships within the genus. Second, allozyme data for 14 presumptive nuclear loci, from 463 individuals sampled from 60 populations, were collected and analysed to assess population similarity or disparity, as well as potential reproductive isolation. The DNA sequence data grouped all populations into seven major monophyletic groups, and the allozyme data provided evidence for reproductive isolation among four of the seven groups, thereby supporting the species status of these groups. The allozyme data suggested that two of the other groups share the same gene pool, and therefore belong to a single species. Finally, the allozyme data revealed two reproductively isolated units within the seventh group, which we suggest represents a case of ‘budding speciation’ based on the DNA gene tree. In total, seven species of the genus Batrachuperus were defined, two of which were previously unknown. The phylogeographical analysis also revealed that vicariance events might have dominated the evolutionary history of this group, but the speciation events might precede the formation of the existing mountain topology. This study demonstrates the importance of including frequency data from multiple nuclear gene loci in determining species boundaries.

Past and present: Phylogeography of the Bufo gargarizans species complex inferred from multi-loci allele sequence and frequency data

Using multi-locus DNA sequence and frequency data, we examined the phylogeographic patterns of the Asian toad Bufo gargarizans species complex. A total of 166 individuals from 56 sites were genotyped for one mitochondrial locus (516 base pairs, ND2 gene) and five nuclear intron loci (Sox9-2, Rho-3, CCNB2-3, UCH-2, DBI-2; 250-350 base pairs each). We conducted a phylogenetic analysis of the allele sequence data to construct gene trees for each individual locus using Bayesian inference, a multi-locus phylogeny based on all five nuclear loci using POFAD, and a phylogenetic network using the NeighborNet algorithm. Furthermore, we used population-genetic analysis of the allele frequency data to reveal ongoing processes, including an analysis of molecular variance (AMOVA), a Bayesian assignment analysis, and a non-equilibrium Bayesian method for estimating recent migration rate. Our phylogenetic analyses showed that the observed divergence in the B. gargarizans species complex likely dated back to 7-8 million years ago. Repeated range expansions during the inter-glacial periods of Pleistocene likely established the current distribution of genetic diversity, although historical vicariant events were still evidenced. Both the west and the southeast regions may have served as refugia during the glaciation, and the range expansion was in general from west to northeast. Additionally, we detected strong ongoing migration both from west to the other regions and from south to north, which likely represents a meta-population dynamic that has emerged over the past ∼10,000 years. The morphologically identified species Bufo tibetanus is not genetically diagnosable, and therefore should be considered a junior synonym of B. gargarizans.