Zhenhua Luo

Evolution by selection, recombination, and gene duplication in MHC class I genes of two Rhacophoridae species

BackgroundComparison of major histocompatibility complex (MHC) genes across vertebrate species can reveal molecular mechanisms underlying the evolution of adaptive immunity-related proteins. As the first terrestrial tetrapods, amphibians deserve special attention because of their exposure to probably increased spectrum of microorganisms compared with ancestral aquatic fishes. Knowledge regarding the evolutionary patterns and mechanisms associated with amphibian MHC genes remains limited. The goal of the present study was to isolate MHC class I genes from two Rhacophoridae species (Rhacophorus omeimontis and Polypedates megacephalus) and examine their evolution.ResultsWe identified 27 MHC class I alleles spanning the region from exon 2 to 4 in 38 tree frogs. The available evidence suggests that these 27 sequences all belong to classical MHC class I (MHC Ia) genes. Although several anuran species only display one MHC class Ia locus, at least two or three loci were observed in P. megacephalus and R. omeimontis, indicating that the number of MHC class Ia loci varies among anuran species. Recombination events, which mainly involve the entire exons, played an important role in shaping the genetic diversity of the 27 MHC class Ia alleles. In addition, signals of positive selection were found in Rhacophoridae MHC class Ia genes. Amino acid sites strongly suggested by program to be under positive selection basically accorded with the putative antigen binding sites deduced from crystal structure of human HLA. Phylogenetic relationships among MHC class I alleles revealed the presence of trans-species polymorphisms.ConclusionsIn the two Rhacophoridae species (1) there are two or three MHC class Ia loci; (2) recombination mainly occurs between the entire exons of MHC class Ia genes; (3) balancing selection, gene duplication and recombination all contribute to the diversity of MHC class Ia genes. These findings broaden our knowledge on the evolution of amphibian MHC systems.

Comparative transcriptome analyses of seven anurans reveal functions and adaptations of amphibian skin

Animal skin, which is the tissue that directly contacts the external surroundings, has evolved diverse functions to adapt to various environments. Amphibians represent the transitional taxon from aquatic to terrestrial life. Exploring the molecular basis of their skin function and adaptation is important to understand the survival and evolutionary mechanisms of vertebrates. However, comprehensive studies on the molecular mechanisms of skin functions in amphibians are scarce. In this study, we sequenced the skin transcriptomes of seven anurans belonging to three families and compared the similarities and differences in expressed genes and proteins. Unigenes and pathways related to basic biological processes and special functions, such as defense, immunity, and respiration, were enriched in functional annotations. A total of 108 antimicrobial peptides were identified. The highly expressed genes were similar in species of the same family but were different among families. Additionally, the positively selected orthologous groups were involved in biosynthesis, metabolism, immunity, and defense processes. This study is the first to generate extensive transcriptome data for the skin of seven anurans and provides unigenes and pathway candidates for further studies on amphibian skin function and adaptation.

Conservation of Terrestrial Vertebrates in a Global Hotspot of Karst Area in Southwestern China.

The karst area of southwest China (KASC) is the largest piece of karst landscape on the earth and a global biodiversity hot-spot with high concentrations of endemic species. Although a number of nature reserves (NRs) have been established across the region, the representativeness of biodiversity of the NR system is still unknown. Based on comprehensive literature and field surveys, and intensive consultations with zoologists and wildlife managers, we compiled distributions of 1,204 terrestrial vertebrate species and 271 NRs in KASC. We found Jinxiu, Mengla, Hekou, and Jinghong have the richest amphibian species; Jinxiu has the highest species richness of reptiles; Jinghong, Menghai, and Mengla have the largest numbers of avian species; whereas, Mengla, Longzhou, and Ningming have the greatest mammalian diversity in the region. Gap analysis among NR system, species richness pattern, and five biogeographic indicators found insufficient representation of the NR system on territorial vertebrate diversity. The conservation effectiveness in Guizhou Province was much lower than that in Guangxi and Yunnan Provinces. Under-representation and over-representation simultaneously occurred in many of the ecoregions, elevation classes, vegetation types, landcover categories, and human disturbance intensity gradients. For conservation of terrestrial vertebrates in KASC, several suggestions were presented in this study.

Geologic events coupled with Pleistocene climatic oscillations drove genetic variation of Omei treefrog (Rhacophorus omeimontis) in southern China

BackgroundPleistocene climatic oscillations and historical geological events may both influence current patterns of genetic variation, and the species in southern China that faced unique climatic and topographical events have complex evolutionary histories. However, the relative contributions of climatic oscillations and geographical events to the genetic variation of these species remain undetermined. To investigate patterns of genetic variation and to test the hypotheses about the factors that shaped the distribution of this genetic variation in species of southern China, mitochondrial genes (cytochrome b and NADH dehydrogenase subunit 2) and nine microsatellite loci of the Omei tree frog (Rhacophorus omeimontis) were amplified in this study.ResultsThe genetic diversity in the populations of R. omeimontis was high. The phylogenetic trees reconstructed from the mitochondrial DNA (mtDNA) haplotypes and the Bayesian genetic clustering analysis based on microsatellite data both revealed that all populations were divided into three lineages (SC, HG and YN). The two most recent splitting events among the lineages coincided with recent geological events (including the intense uplift of the Qinghai-Tibet Plateau, QTP and the subsequent movements of the Yun-Gui Plateau, YGP) and the Pleistocene glaciations. Significant expansion signals were not detected in mismatch analyses or neutrality tests. And the effective population size of each lineage was stable during the Pleistocene.ConclusionsBased on the results of this study, complex geological events (the recent dramatic uplift of the QTP and the subsequent movements of the YGP) and the Pleistocene glaciations were apparent drivers of the rapid divergence of the R. omeimontis lineages. Each diverged lineages survived in situ with limited gene exchanges, and the stable demographics of lineages indicate that the Pleistocene climatic oscillations were inconsequential for this species. The analysis of genetic variation in populations of R. omeimontis contributes to the understanding of the effects of changes in climate and of geographical events on the dynamic development of contemporary patterns of genetic variation in the species of southern China.

Impacts of climate change on the distribution of Sichuan snub-nosed monkeys (Rhinopithecus roxellana) in Shennongjia area, China.

Understanding the effects of climate change on primate ranging patterns is crucial for conservation planning. Rhinopithecus roxellana is an endangered primate species distributed in mountainous forests at the elevation of 1500–3500 m a.s.l. in China. Our study site, the Shennongjia National Nature Reserve, represents the eastern‐most distribution of this species. This area has experienced significant habitat loss and fragmentation because of human population growth, increased farming and logging, and climate change. To estimate how changes in temperature and rainfall will affect the presumed future distribution of this species, we examined eco‐geographic factors including bioclimate, habitat (vegetation type, landcover, etc.), topography, and human impact (human population, gross domestic product, etc.), and provide suggestions for management and conservation. We used a maximum entropy approach to predict the location and distribution of habitats suitable for R. roxellana in the present, 2020, 2050, and 2080 based on 33 environmental parameters, three general circulation models, three emissions scenarios, and two dispersal hypotheses. According to the ensemble modeling, we found range reductions of almost 30% by 2020, 70% by 2050, and over 80% by 2080. Although no obvious differences were found in distribution change based on full and zero dispersal assumptions, our results revealed range reductions in response to elevational, latitudinal, and longitudinal gradients, with the monkeys forced to migrate to higher elevations over time. Bioclimte factors, such as temperature, precipitation, evapo‐transpiration, and aridity condition, were dominant contributors to range shifting. As habitat loss due to human influence and climate change is likely to be even more severe in the future, we considered three conservation hot‐spots in the Shennongjia area and recommended: (i) securing existing reserves and establishing new reserves, (ii) re‐designing management systems to include the Shenongjia reserve and the surrounding reserves and highlighting ecosystem protection at higher elevations, and (iii) using finer‐scale research to guide the conservation planning and education in order to enhance protection and awareness in the local community. National and provincial conservation policies should integrate projections of climate change in making effective conservation strategies. Am. J. Primatol. 77:135–151, 2015.

Male-male competition drives sexual selection and group spawning in the Omei treefrog, Rhacophorus omeimontis

Male-male competition and female mate choice based on ornamentation and genetic traits are the main drivers of animal sexual selection and group spawning. If male-male competition is intense, males with specific phenotypes should have advantages in breeding success or occupying superior mating positions. If female choice is important, females should have preferences for mate relatedness or males with good genes or optimal genetic compatibility against themselves. To detect the intensities of male-male competition and female choice and test the good genes and genetic compatibility hypotheses, we observed breeding behaviors, measured individual body lengths and breeding success indicators, and calculated male genetic heterozygosity, male–female relatedness, and genetic dissimilarity in an Omei treefrog (Rhacophorus omeimontis) population in Badagongshan, China. Our analyses showed that larger males obtained larger mates, had more mating opportunities, occupied better amplectant positions, and produced more offspring. However, females showed no inbreeding/outbreeding bias in mate choice, and the good genes and genetic compatibility hypotheses were not supported in female selection on mates and amplectant positions. We considered male-male competition as the main driver of sexual selection and group spawning in this prolonged mating species because the cost of choosing mates with specific genetic traits may be high for females.Significance statementMale-male competition and female mate choice are the main drivers of animal sexual selection and mating behaviors. However, their impacts on the evolution of mating system are not yet quite clear. By studying the Omei treefrog, a Chinese endemic anuran species with common group spawning behaviors, this research posed the importance of male-male competition and female mate choice and discussed the mechanisms of sexual selection and multiple mating, which are the most essential and debated issues in evolutionary biology and behavioral ecology. We found that, in this prolonged mating and lek-patterned species, male-male competition is the main driver of sexual selection and group spawning. Larger males can get larger females, have more breeding opportunities, occupy better amplectant positions, and thus, obtain greater numbers of offspring. Whereas, females have no significant preferences on ornamentation and genetic traits of their males or mating positions, and the inferior males “make the best of a bad lot” by joining mating pairs to produce mating groups. Our study provides empirical evidence of reproductive mechanisms of amphibian species and could advance the understandings on the evolution of animals’ sexual selection and mating system.

Assessment of Habitat Fragmentation and Corridors for an Isolated Subspecies of the Sichuan Golden Snub-Nosed Monkey, Rhinopithecus roxellana hubeiensis

Understanding habitat quality and landscape connectivity and exploring corridors connecting habitat patches are crucial for conservation, particularly for species distributed among isolated populations. The Sichuan golden snub-nosed monkey, Rhinopithecus roxellana, is an Endangered primate species endemic to mountainous forests in China. Its easternmost distribution lies in the Shennongjia area, which harbors an isolated subspecies, R. roxellana hubeiensis. Unfortunately, it has experienced significant habitat loss, fragmentation, and dramatic population decline in recent decades, primarily due to increased human disturbance. To quantify habitat quality, identify suitable habitat patches, and detect possible linkages among these patches for R. roxellana hubeiensis, we conducted habitat suitability assessments and landscape connectivity analyses in the Shennongjia area based on a set of environmental factors. We created a habitat quality model and a movement cost surface for the Shennongjia area based on a habitat suitability index, graph theory, expert knowledge, field experience, and information from the literature. Our results show that suitable habitat for R. roxellana hubeiensis in Shennongjia is fragmented and limited, and that this is particularly true for highly suitable habitats. We detected six core habitat patches and six least-cost paths and corridors. Our study does not provide accurate distributions of the monkeys and their habitat use. However, it identifies the most feasible and traversable habitats and corridors, which should be conservation priorities for this subspecies, and provides valuable guidance for reevaluating habitat conservation plans.

High genetic diversity in the endangered and narrowly distributed amphibian species Leptobrachium leishanense.

Threatened species typically have a small or declining population size, which make them highly susceptible to loss of genetic diversity through genetic drift and inbreeding. Genetic diversity determines the evolutionary potential of a species; therefore, maintaining the genetic diversity of threatened species is essential for their conservation. In this study, we assessed the genetic diversity of the adaptive major histocompatibility complex (MHC) genes in an endangered and narrowly distributed amphibian species, Leptobrachium leishanense in Southwest China. We compared the genetic variation of MHC class I genes with that observed in neutral markers (5 microsatellite loci and cytochrome b gene) to elucidate the relative roles of genetic drift and natural selection in shaping the current MHC polymorphism in this species. We found a high level of genetic diversity in this population at both MHC and neutral markers compared with other threatened amphibian species. Historical positive selection was evident in the MHC class I genes. The higher allelic richness in MHC markers compared with that of microsatellite loci suggests that selection rather than genetic drift plays a prominent role in shaping the MHC variation pattern, as drift can affect all the genome in a similar way but selection directly targets MHC genes. Although demographic analysis revealed no recent bottleneck events in L. leishanense, additional population decline will accelerate the dangerous status for this species. We suggest that the conservation management of L. leishanense should concentrate on maximizing the retention of genetic diversity through preventing their continuous population decline. Protecting their living habitats and forbidding illegal hunting are the most important measures for conservation of L. leishanense.