Evolutionary Conservation Genomics Reveals Recent Speciation and Local Adaptation in Threatened Takins.

Incorrect species delimitation will lead to inappropriate conservation decisions, especially for threatened species. The takin (Budorcas taxicolor) is a large artiodactyl endemic to the Himalayan-Hengduan-Qinling Mountains and is well known for its threatened status and peculiar appearance. However, the speciation, intraspecies taxonomy, evolutionary history, and adaptive evolution of this species still remain unclear, which greatly hampers its scientific conservation. Here, we de novo assembled a high-quality chromosome-level genome of takin and resequenced the genomes of 75 wild takins. Phylogenomics revealed that takin was positioned at the root of Caprinae. Population genomics based on the autosome, X chromosome, and Y chromosome SNPs and mitochondrial genomes consistently revealed the existence of two phylogenetic species and recent speciation in takins: the Himalayan takin (B. taxicolor) and the Chinese takin (B. tibetana), with the support of morphological evidence. Two genetically divergent subspecies were identified in both takin species, rejecting three previously proposed taxonomical viewpoints. Furthermore, their distribution boundaries were determined, suggesting that large rivers play important roles in shaping the genetic partition. Compared with the other subspecies, the Qinling subspecies presented the lowest genomic diversity, higher linkage disequilibrium, inbreeding, and genetic load, thus is in urgent need of genetic management and protection. Moreover, coat color gene (PMEL) variation may be responsible for the adaptive coat color difference between the two species following Gloger's rule. Our findings provide novel insights into the recent speciation, local adaptation, scientific conservation of takins, and biogeography of the Himalaya-Hengduan biodiversity hotspot.

Genomic Signatures of Coevolution between Nonmodel Mammals and Parasitic Roundworms.

Antagonistic coevolution between host and parasite drives species evolution. However, most of the studies only focus on parasitism adaptation and do not explore the coevolution mechanisms from the perspective of both host and parasite. Here, through the de novo sequencing and assembly of the genomes of giant panda roundworm, red panda roundworm, and lion roundworm parasitic on tiger, we investigated the genomic mechanisms of coevolution between nonmodel mammals and their parasitic roundworms and those of roundworm parasitism in general. The genome-wide phylogeny revealed that these parasitic roundworms have not phylogenetically coevolved with their hosts. The CTSZ and prolyl 4-hydroxylase subunit beta (P4HB) immunoregulatory proteins played a central role in protein interaction between mammals and parasitic roundworms. The gene tree comparison identified that seven pairs of interactive proteins had consistent phylogenetic topology, suggesting their coevolution during host-parasite interaction. These coevolutionary proteins were particularly relevant to immune response. In addition, we found that the roundworms of both pandas exhibited higher proportions of metallopeptidase genes, and some positively selected genes were highly related to their larvae's fast development. Our findings provide novel insights into the genetic mechanisms of coevolution between nonmodel mammals and parasites and offer the valuable genomic resources for scientific ascariasis prevention in both pandas.

A genome-wide scan for copy number variations using high-density single nucleotide polymorphism array in Simmental cattle.

Copy number variations (CNVs) have recently been identified as promising sources of genetic variation, complementary to single nucleotide polymorphisms (SNPs). As a result, detection of CNVs has attracted a great deal of attention. In this study, we performed genome-wide CNV detection using Illumina Bovine HD BeadChip (770k) data on 792 Simmental cattle. A total of 263 CNV regions (CNVRs) were identified, which included 137 losses, 102 gains and 24 regions classified as both loss and gain, covering 35.48 Mb (1.41%) of the bovine genome. The length of these CNVRs ranged from 10.18 kb to 1.76 Mb, with an average length of 134.78 kb and a median length of 61.95 kb. In 136 of these regions, a total of 313 genes were identified related to biological functions such as transmembrane activity and olfactory transduction activity. To validate the results, we performed quantitative PCR to detect nine randomly selected CNVRs and successfully confirmed seven (77.6%) of them. Our results present a map of cattle CNVs derived from high-density SNP data, which expands the current CNV map of the cattle genome and provides useful information for investigation of genomic structural variation in cattle.

Geographical patterns and determinants of insect biodiversity in China.

Insects play important roles in the maintenance of ecosystem functioning and the provision of livelihoods for millions of people. However, compared with terrestrial vertebrates and angiosperms, such as the giant panda, crested ibis, and the metasequoia, insect conservation has not attracted enough attention, and a basic understanding of the geographical biodiversity patterns for major components of insects in China is lacking. Herein, we investigated the geographical distribution of insect biodiversity across multiple dimensions (taxonomic, genetic, and phylogenetic diversity) based on the spatial distribution and molecular DNA sequencing data of insects. Our analysis included 18 orders, 360 families, 5,275 genera, and 14,115 species of insects. The results revealed that Southwestern and Southeastern China harbored higher insect biodiversity and numerous older lineages, representing a museum, whereas regions located in Northwestern China harbored lower insect biodiversity and younger lineages, serving as an evolutionary cradle. We also observed that mean annual temperature and precipitation had significantly positive effects, whereas altitude had significantly negative effects on insect biodiversity in most cases. Moreover, cultivated vegetation harbored the highest insect taxonomic and phylogenetic diversity, and needleleaf and broadleaf mixed forests harbored the highest insect genetic diversity. These results indicated that human activities may positively contribute to insect spatial diversity on a regional scale. Our study fills a knowledge gap in insect spatial diversity in China. These findings could help guide national-level conservation plans and the post-2020 biodiversity conservation framework.

The genome of African manatee Trichechus senegalensis reveals secondary adaptation to the aquatic environment.

Sirenians exhibit unique aquatic adaptations, showcasing both convergent adaptive features shared with cetaceans and unique characteristics such as cold sensitivity and dense bones. Here, we report a chromosome-level genome of the African manatee (Trichechus senegalensis) with high continuity, completeness, and accuracy. We found that genes associated with osteopetrosis have undergone positive selection (CSF1R and LRRK1) or pseudogenized (FAM111A and IGSF23) in the African manatee, potentially contributing to the dense bone formation. The loss of KCNK18 may have increased their sensitivity to cold water temperatures. Moreover, we identified convergent evolutionary signatures in 392 genes among fully aquatic mammals, primarily enriched in skin or skeletal system development and circadian rhythm, which contributed to the transition from terrestrial to fully aquatic lifestyles. The African manatee currently possesses a small effective population size and low genome-wide heterozygosity. Overall, our study provides genetic resources for understanding the evolutionary characteristics and conservation efforts of this species.

Conservation genomics of the critically endangered Chinese pangolin.

The Chinese pangolin (Manis pentadactyla, MP) has been extensively exploited and is now on the brink of extinction, but its population structure, evolutionary history, and adaptive potential are unclear. Here, we analyzed 94 genomes from three subspecies of the Chinese pangolin and identified three distinct genetic clusters (MPA, MPB, and MPC), with MPB further divided into MPB1 and MPB2 subpopulations. The divergence of these populations was driven by past climate change. For MPB2 and MPC, recent human activities have caused dramatic population decline and small population size as well as increased inbreeding, but not decrease in genomic variation and increase in genetic load probably due to strong gene flow; therefore, it is crucial to strengthen in situ habitat management for these two populations. By contrast, although human activities have a milder impact on MPA, it is at high risk of extinction due to long-term contraction and isolation, and genetic rescue is urgently needed. MPB1 exhibited a relatively healthy population status and can potentially serve as a source population. Overall, our findings provide novel insights into the conservation of the Chinese pangolin and biogeography of the mammals of eastern Asia.

Influence of Last Glacial Maximum legacies on functional diversity and community assembly of extant Chinese terrestrial vertebrates.

Contemporary biodiversity patterns are shaped by not only modern climate but also factors such as past climate fluctuations. Investigating the relative degree of paleoclimate legacy could help us understand the formation of current biodiversity patterns. However, an assessment of this issue in China is lacking. Here, we investigated the phylogenetic structure and functional diversity patterns of Chinese terrestrial vertebrates. We found that Southern China harbored higher functional richness, while Northern and Western China were more phylogenetically clustered with higher functional divergence and evenness, indicating environmental filtering effects. Moreover, we found that drastic Last Glacial Maximum climate changes were positively related to phylogenetic clustering, lower functional richness, and higher functional divergence and evenness, although this effect varied among different taxonomic groups. We further found that mammal communities experiencing more drastic Last Glacial Maximum temperature changes were characterized by "faster" life-history trait values. Our findings provide new evidence of the paleoclimate change legacies influencing contemporary biodiversity patterns that will help guide national-level conservation plans.

Conservation priorities for global marine biodiversity across multiple dimensions.

Marine biodiversity plays important roles in ocean ecosystem services and has substantial economic value. Species diversity, genetic diversity and phylogenetic diversity, which reflect the number, evolutionary potential and evolutionary history of species in ecosystem functioning, are three important dimensions of biodiversity. Marine-protected areas have been demonstrated as an effective area-based tool for protecting marine biodiversity, but only 2.8% of the ocean has been fully protected. It is urgent to identify global conservation priority areas and percentage of the ocean across multiple dimensions of biodiversity based on Post-2020 Global Biodiversity Framework. Here, we investigate the spatial distribution of marine genetic and phylogenetic diversity using 80 075 mitochondrial DNA barcode sequences from 4316 species and a newly constructed phylogenetic tree of 8166 species. We identify that the Central Indo-Pacific Ocean, Central Pacific Ocean and Western Indian Ocean harbor high levels of biodiversity across three dimensions of biodiversity, which could be designated as conservation priority areas. We also find that strategically protecting ∼22% of the ocean would allow us to reach the target of conserving ∼95% of currently known taxonomic, genetic and phylogenetic diversity. Our study provides insights into the spatial distribution pattern of multiple marine diversities and the findings would help to design comprehensive conservation schemes for global marine biodiversity.

The rough-toothed dolphin genome provides new insights into the genetic mechanism of its rough teeth.

The rough-toothed dolphin (Steno bredanensis) is characterized by having teeth covered in finely wrinkled vertical ridges, which is a general manifestation of amelogenesis imperfecta. The rough surfaces are hypothesized to be an evolutionary morphological trait of feeding adaptation to increase the dolphin's grip on prey. Here, we assembled a rough-toothed dolphin genome and performed the comparative genomic analysis to reveal the genetic basis of the special enamel. Results showed that genes related to enamel development or dental diseases have undergone diversified adaptive changes that may shape the special enamel morphology of this dolphin species, including positive selection (CLDN19, PRKCE, SSUH2, and WDR72), rapid evolution (LAMB3), or unique amino acid substitutions (AMTN, ENAM, MMP20, and KLK4). Meanwhile, the historical demography of rough-toothed dolphin indicated several distinct population fluctuations associated with climate change. The genome-wide heterozygosity of this dolphin is in the middle of all published data for cetaceans. Although the population is considerable, there may be population or subspecies differentiation, and with the global warming and the increasing disturbance of human activities, we should pay more attention to protection in the future. Together, our study brings new insights into the genetic mechanisms that may have driven the evolution of the special enamel morphology in rough-toothed dolphins and provides the first results of genetic heterozygosity and population historical dynamics of this species, which have important guiding implications for the conservation of this dolphin species.

[Artificial selection for cattle based on high-density SNP markers].

With the implementation of genetic improvement in recent years, artificial selection has greatly improved beef cattle production performance and its genetic basis has been dramatically changed. In this study, based on the Illumina BovineSNP50 (54K) and BovineHD (770K) BeadChip and the FST value, we analyzed the genetic differentiation of cattle and screened the imprints of selection in bovine genome. Finally, we found 47104 OUTLIER SNP loci and 3064 candidate genes, for example, CLIC5, TG, CACNA2D1, and FSHR etc. The biological processes and molecular functions of genes were analyzed through gene annotation.The results of this study established a genome-wide map of selection footprints in beef cattle genome and a clue for in-depth study of artificial selection and understanding of biological evolution.Our results indicate that artificial selection has played an important role in cattle breed genetic improvement.

Spatial patterns and conservation of genetic and phylogenetic diversity of wildlife in China.

Genetic diversity and phylogenetic diversity reflect the evolutionary potential and history of species, respectively. However, the levels and spatial patterns of genetic and phylogenetic diversity of wildlife at the regional scale have largely remained unclear. Here, we performed meta-analyses of genetic diversity in Chinese terrestrial vertebrates based on three genetic markers and investigated their phylogenetic diversity based on a dated phylogenetic tree of 2461 species. We detected strong positive spatial correlations among mitochondrial DNA-based genetic diversity, phylogenetic diversity, and species richness. Moreover, the terrestrial vertebrates harbored higher genetic and phylogenetic diversity in South China and Southwest China than in other regions. Last, climatic factors (precipitation and temperature) had significant positive effects while altitude and human population density had significant negative impacts on levels of mitochondrial DNA-based genetic diversity in most cases. Our findings will help guide national-level genetic diversity conservation plans and a post-2020 biodiversity conservation framework.

Molecular mechanisms and topological consequences of drastic chromosomal rearrangements of muntjac deer.

Muntjac deer have experienced drastic karyotype changes during their speciation, making it an ideal model for studying mechanisms and functional consequences of mammalian chromosome evolution. Here we generated chromosome-level genomes for Hydropotes inermis (2n = 70), Muntiacus reevesi (2n = 46), female and male M. crinifrons (2n = 8/9) and a contig-level genome for M. gongshanensis (2n = 8/9). These high-quality genomes combined with Hi-C data allowed us to reveal the evolution of 3D chromatin architectures during mammalian chromosome evolution. We find that the chromosome fusion events of muntjac species did not alter the A/B compartment structure and topologically associated domains near the fusion sites, but new chromatin interactions were gradually established across the fusion sites. The recently borne neo-Y chromosome of M. crinifrons, which underwent male-specific inversions, has dramatically restructured chromatin compartments, recapitulating the early evolution of canonical mammalian Y chromosomes. We also reveal that a complex structure containing unique centromeric satellite, truncated telomeric and palindrome repeats might have mediated muntjacs' recurrent chromosome fusions. These results provide insights into the recurrent chromosome tandem fusion in muntjacs, early evolution of mammalian sex chromosomes, and reveal how chromosome rearrangements can reshape the 3D chromatin regulatory conformations during species evolution.

Genomic evidence for two phylogenetic species and long-term population bottlenecks in red pandas.

The red panda (Ailurus fulgens), an endangered Himalaya-endemic mammal, has been classified as two subspecies or even two species - the Himalayan red panda (A. fulgens) and the Chinese red panda (Ailurus styani) - based on differences in morphology and biogeography. However, this classification has remained controversial largely due to lack of genetic evidence, directly impairing scientific conservation management. Data from 65 whole genomes, 49 Y-chromosomes, and 49 mitochondrial genomes provide the first comprehensive genetic evidence for species divergence in red pandas, demonstrating substantial inter-species genetic divergence for all three markers and correcting species-distribution boundaries. Combined with morphological evidence, these data thus clearly define two phylogenetic species in red pandas. We also demonstrate different demographic trajectories in the two species: A. styani has experienced two population bottlenecks and one large population expansion over time, whereas A. fulgens has experienced three bottlenecks and one very small expansion, resulting in very low genetic diversity, high linkage disequilibrium, and high genetic load.

Chromosome-level genome assembly for giant panda provides novel insights into Carnivora chromosome evolution.

BACKGROUND: Chromosome evolution is an important driver of speciation and species evolution. Previous studies have detected chromosome rearrangement events among different Carnivora species using chromosome painting strategies. However, few of these studies have focused on chromosome evolution at a nucleotide resolution due to the limited availability of chromosome-level Carnivora genomes. Although the de novo genome assembly of the giant panda is available, current short read-based assemblies are limited to moderately sized scaffolds, making the study of chromosome evolution difficult. RESULTS: Here, we present a chromosome-level giant panda draft genome with a total size of 2.29 Gb. Based on the giant panda genome and published chromosome-level dog and cat genomes, we conduct six large-scale pairwise synteny alignments and identify evolutionary breakpoint regions. Interestingly, gene functional enrichment analysis shows that for all of the three Carnivora genomes, some genes located in evolutionary breakpoint regions are significantly enriched in pathways or terms related to sensory perception of smell. In addition, we find that the sweet receptor gene TAS1R2, which has been proven to be a pseudogene in the cat genome, is located in an evolutionary breakpoint region of the giant panda, suggesting that interchromosomal rearrangement may play a role in the cat TAS1R2 pseudogenization. CONCLUSIONS: We show that the combined strategies employed in this study can be used to generate efficient chromosome-level genome assemblies. Moreover, our comparative genomics analyses provide novel insights into Carnivora chromosome evolution, linking chromosome evolution to functional gene evolution.

Synteny search identifies carnivore Y chromosome for evolution of male specific genes.

The explosive accumulation of mammalian genomes has provided a valuable resource to characterize the evolution of the Y chromosome. Unexpectedly, the Y-chromosome sequence has been characterized in only a small handful of species, with the majority being model organisms. Thus, identification of Y-linked scaffolds from unordered genome sequences is becoming more important. Here, we used a syntenic-based approach to generate the scaffolds of the male-specific region of the Y chromosome (MSY) from the genome sequence of 6 male carnivore species. Our results identified 14, 15, 9, 28, 14 and 11 Y-linked scaffolds in polar bears, pacific walruses, red pandas, cheetahs, ferrets and tigers, covering 1.55 Mbp, 2.62 Mbp, 964 Kb, 1.75 Mb, 2.17 Mbp and 1.84 Mb MSY, respectively. All the candidate Y-linked scaffolds in 3 selected species (red pandas, polar bears and tigers) were successfully verified using polymerase chain reaction. We re-annotated 8 carnivore MSYs including these 6 Y-linked scaffolds and domestic dog and cat MSY; a total of 11 orthologous genes conserved in at least 7 of the 8 carnivores were identified. These 11 Y-linked genes have significantly higher evolutionary rates compared with their X-linked counterparts, indicating less purifying selection for MSY genes. Taken together, our study shows that the approach of synteny search is a reliable and easily affordable strategy to identify Y-linked scaffolds from unordered carnivore genomes and provides a preliminary evolutionary study for carnivore MSY genes.

Conservation genetics and genomics of threatened vertebrates in China.

Conservation genetics and genomics are two independent disciplines that focus on using new techniques in genetics and genomics to solve problems in conservation biology. During the past two decades, conservation genetics and genomics have experienced rapid progress. Here, we summarize the research advances in the conservation genetics and genomics of threatened vertebrates (e.g., carnivorans, primates, ungulates, cetaceans, avians, amphibians and reptiles) in China. First, we introduce the concepts of conservation genetics and genomics and their development. Second, we review the recent advances in conservation genetics research, including noninvasive genetics and landscape genetics. Third, we summarize the progress in conservation genomics research, which mainly focuses on resolving genetic problems relevant to conservation such as genetic diversity, genetic structure, demographic history, and genomic evolution and adaptation. Finally, we discuss the future directions of conservation genetics and genomics.

Searching for new loci and candidate genes for economically important traits through gene-based association analysis of Simmental cattle.

Single-marker genome-wide association study (GWAS) is a convenient strategy of genetic analysis that has been successful in detecting the association of a number of single-nucleotide polymorphisms (SNPs) with quantitative traits. However, analysis of individual SNPs can only account for a small proportion of genetic variation and offers only limited knowledge of complex traits. This inadequacy may be overcome by employing a gene-based GWAS analytic approach, which can be considered complementary to the single-SNP association analysis. Here we performed an initial single-SNP GWAS for bone weight (BW) and meat pH value with a total of 770,000 SNPs in 1141 Simmental cattle. Additionally, 21836 cattle genes collected from the Ensembl Genes 83 database were analyzed to find supplementary evidence to support the importance of gene-based association study. Results of the single SNP-based association study showed that there were 11 SNPs significantly associated with bone weight (BW) and two SNPs associated with meat pH value. Interestingly, all of these SNPs were located in genes detected by the gene-based association study.