Historical RNA expression profiles from the extinct Tasmanian tiger

Paleogenomics continues to yield valuable insights into the evolution, population dynamics, and ecology of our ancestors and other extinct species. However, DNA sequencing cannot reveal tissue-specific gene expression, cellular identity, or gene regulation, which are only attainable at the transcriptional level. Pioneering studies have shown that useful RNA can be extracted from ancient specimens preserved in permafrost and historical skins from extant canids, but no attempts have been made so far on extinct species. We extract, sequence, and analyze historical RNA from muscle and skin tissue of a ∼130-year-old Tasmanian tiger (Thylacinus cynocephalus) preserved in desiccation at room temperature in a museum collection. The transcriptional profiles closely resemble those of extant species, revealing specific anatomical features such as slow muscle fibers or blood infiltration. Metatranscriptomic analysis, RNA damage, tissue-specific RNA profiles, and expression hotspots genome-wide further confirm the thylacine origin of the sequences. RNA sequences are used to improve proteincoding and noncoding annotations, evidencing missing exonic loci and the location of ribosomal RNA genes while increasing the number of annotated thylacine microRNAs from 62 to 325. We discover a thylacine-specific microRNA isoform that could not have been confirmed without RNA evidence. Finally, we detect traces of RNA viruses, suggesting the possibility of profiling viral evolution. Our results represent the first successful attempt to obtain transcriptional profiles from an extinct animal species, providing thought-to-be-lost information on gene expression dynamics. These findings hold promising implications for the study of RNA molecules across the vast collections of natural history museums and from well-preserved permafrost remains

Integrating multi-taxon palaeogenomes and sedimentary ancient DNA to study past ecosystem dynamics

Ancient DNA (aDNA) has played a major role in our understanding of the past. Important advances in the sequencing and analysis of aDNA from a range of organisms have enabled a detailed understanding of processes such as past demography, introgression, domestication, adaptation and speciation. However, to date and with the notable exception of microbiomes and sediments, most aDNA studies have focused on single taxa or taxonomic groups, making the study of changes at the community level challenging. This is rather surprising because current sequencing and analytical approaches allow us to obtain and analyse aDNA from multiple source materials. When combined, these data can enable the simultaneous study of multiple taxa through space and time, and could thus provide a more comprehensive understanding of ecosystem-wide changes. It is therefore timely to develop an integrative approach to aDNA studies by combining data from multiple taxa and substrates. In this review, we discuss the various applications, associated challenges and future prospects of such an approach

Grey wolf genomic history reveals a dual ancestry of dogs

The grey wolf (Canis lupus) was the first species to give rise to a domestic population, and they remained widespread throughout the last Ice Age when many other large mammal species went extinct. Little is known, however, about the history and possible extinction of past wolf populations or when and where the wolf progenitors of the present-day dog lineage (Canisfamiliaris) lived(1-8). Here we analysed 72 ancient wolf genomes spanning the last 100,000 years from Europe, Siberia and North America. We found that wolf populations were highly connected throughout the Late Pleistocene, with levels of differentiation an order of magnitude lower than they are today. This population connectivity allowed us to detect natural selection across the time series, including rapid fixation of mutations in the gene IFT8840,000-30,000 years ago. We show that dogs are overall more closely related to ancient wolves from eastern Eurasia than to those from western Eurasia, suggesting a domestication process in the east. However, we also found that dogs in the Near East and Africa derive up to half of their ancestry from a distinct population related to modern southwest Eurasian wolves, reflecting either an independent domestication process or admixture from local wolves. None of the analysed ancient wolf genomes is a direct match for either of these dog ancestries, meaning that the exact progenitor populations remain to be located.Peer reviewe

Origins and genetic legacy of prehistoric dogs

Dogs were the first domestic animal, but little is known about their population history and to what extent it was linked to humans. We sequenced 27 ancient dog genomes and found that all dogs share a common ancestry distinct from present-day wolves, with limited gene flow from wolves since domestication but substantial dog-to-wolf gene flow. By 11,000 years ago, at least five major ancestry lineages had diversified, demonstrating a deep genetic history of dogs during the Paleolithic. Coanalysis with human genomes reveals aspects of dog population history that mirror humans, including Levant-related ancestry in Africa and early agricultural Europe. Other aspects differ, including the impacts of steppe pastoralist expansions in West and East Eurasia and a near-complete turnover of Neolithic European dog ancestry

Large carnivore population turnover and ecological change during the Late Quaternary

The cave lion (Panthera spelaea), the grey wolf (Canis lupus) and the brown bear (Ursus arctos) all shared an intercontinental distribution across the northern hemisphere during most of the Late Quaternary, and experienced repeated events of climate change. The cave lion went extinct at the end of the Pleistocene and although the wolf and the bear have survived until present day, recent human persecution has caused demographic bottlenecks and local extinctions. In this thesis, mitochondrial and nuclear DNA was analyzed from ancient and modern samples in order to study spatiotemporal changes in genetic diversity in the three species. Mitochondrial sequences analyzed from 48 radiocarbon dated cave lion remains revealed two haplogroups, of which the more genetically diverse seemingly disappeared around 41,000 years BP. Serial coalescent simulations on the data supported a population bottleneck in Beringia between roughly 47-18,000 years BP. Its long duration prevents a specific causal factor to be singled out, but the early onset and overlapping declines of other large mammals in the region suggests that major environmental changes greatly impacted the fauna of Beringia during this time. Using a similar genetic marker, a set of 126 modern wolves and two Siberian wolf remains of Late Pleistocene age were analyzed. The sequences yielded from the latter samples pertained to a basal haplogroup, which contained all Late Pleistocene wolves from previous studies. As data from both modern and ancient wolves were combined, a pattern of decreasing genetic diversity was identified around the Pleistocene-Holocene transition. This decrease was further tested by serial coalescent simulations, which supported a bottleneck in northern North America around this time. Further analyses were applied to one of the ancient wolf remains from Siberia, producing a draft genome sequence and a complete mitochondrial genome. Given the radiocarbon date of the Siberian wolf, a slower mutation rate could be inferred, which pushed back the split between the lineages leading to modern wolves and dogs to at least 27,000 years BP. The Siberian wolf was positioned close to the split but basal to these lineages. A global comparison with modern dogs indicated a closer genetic affiliation between the Siberian wolf and some arctic breeds. For the brown bear, phylogeographic changes in Europe were studied over the last 50,000 years, using radiocarbon dating and mitochondrial sequences. When concatenated and compared with published data, the mtDNA revealed a turnover event just before the LGM, while the dating confirmed a presence of brown bears at relatively high latitudes during this period. Marked shifts in population size were also inferred. Furthermore, data of stable isotope levels confirmed a dietary shift to increasing herbivory around the LGM. Finally, a recent anthropogenic bottleneck among Scandinavian brown bears was studied. While no change in genetic structure could be detected, mitochondrial and microsatellite markers showed a decline in genetic diversity, especially pronounced in the southern subpopulation. ABC simulations supported a bottleneck taking place across all of Scandinavia. Taken together, this thesis have identified and elucidated several impacts on genetic diversity in the past populations of large carnivores. The use of different genetic markers has enabled comparisons with published data, but also revealed their comparatively different benefits and limitations. Overall, the presented studies compose a synthesis of past population dynamics in large carnivores, uniquely revealed by ancient DNA.At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 4: Manuscript.</p

Population Demography and Genetic Diversity in the Pleistocene Cave Lion

With a range that covered most of northern Eurasia and parts of North America, the cave lion (Panthera spelaea) was one of the most widespread carnivores of the Late Pleistocene. Earlier ancient DNA analyses have shown that it is distinct from modern lions, and have suggested a demographic decline in Beringia during marine isotope stage 3 (MIS 3). Here, we further investigate the Late Pleistocene population dynamics in more detail by combining a powerful algorithm that couples MCMC with coalescent simulations under an approximate Bayesian computation framework. We use an ancient DNA dataset of previously published (n = 34) and new radiocarbon dated specimens (n = 14). Phylogenetic and network analyses based on the mitochondrial control region and the ATP8 gene identified two major haplogroups, one of which appears to vanish around 41,000 cal a BP. The approximate Bayesian computation analysis suggested a decline in effective population size (Ne) in Beringia of at least a 2-fold magnitude that began approximately 47,000 cal a BP, followed by an increase in Ne, most likely around 18,000 cal a BP. The cave lion went through a demographic bottleneck during MIS 3, which may have lasted for several tens of thousands of years, and only recovered shortly before the species\u27 extinction. Several other large mammal species display similar declines in genetic diversity in Beringia during MIS 3, suggesting that major environmental changes might have affected megafaunal populations during this time period

Contesting Marginality : The Boreal Forest of Middle Scandinavia and the Worlds Outside

In this article, we present ongoing archaeological research into Scandinavia's forested inland region, suggesting that its people and communities were socially and economically integrated into systems of trade and in close interaction with the worlds outside, as early as the first centuries of the Common Era. The article presents a range of archaeological evidence, from ca. 500 to 1400 CE, for processes of ecological globalization, manifested by the exploitation of local landscapes and the extraction of valued products that could be transformed into commodities through crafts and trade. These forested landscapes were reliant on—and also shaped by—complex social and economic relations reflecting interrelated socio-economic systems of extraction, production, and consumption. Our main argument is that these landscapes are crucial to identifying and understanding the contours of the premodern global North