Genetic diversity, demographic history and neo-sex chromosomes in the Critically Endangered Raso lark
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ABSTRACT: Analysis of study genetic diversity, demography and neo-sex chromosomes in the Raso lark Alauda razae and its relatives the Eurasian Skylark A. arvensis, Oriental skylark A. gulgula and crested lark Galerida cristata, based on RAD-seq analysis.
Project description:Small effective population sizes could expose island species to inbreeding and loss of genetic variation. Here, we investigate factors shaping genetic diversity in the Raso lark, which has been restricted to a single islet for approximately 500 years, with a population size of a few hundred. We assembled a reference genome for the related Eurasian skylark and then assessed diversity and demographic history using RAD-seq data (75 samples from Raso larks and two related mainland species). We first identify broad tracts of suppressed recombination in females, indicating enlarged neo-sex chromosomes. We then show that genetic diversity across autosomes in the Raso lark is lower than in its mainland relatives, but inconsistent with long-term persistence at its current population size. Finally, we find that genetic signatures of the recent population contraction are overshadowed by an ancient expansion and persistence of a very large population until the human settlement of Cape Verde. Our findings show how genome-wide approaches to study endangered species can help avoid confounding effects of genome architecture on diversity estimates, and how present-day diversity can be shaped by ancient demographic events.
Project description:Circadian behaviors are regulated by intrinsic biological clocks consisting of central molecular oscillators and output pathways. Despite significant progress in elucidating the central timekeeping mechanisms, the molecular pathways coupling the circadian pacemaker to overt rhythmic behavior and physiology remain elusive. The Drosophila LARK RNA-binding protein is a candidate for such a coupling factor. Previous research indicates that LARK functions downstream of the clock to mediate behavioral outputs. To better understand the roles of LARK in the Drosophila circadian system, we sought to identify RNA molecules associated with LARK in vivo, using a novel strategy that involves capturing the RNA ligands by immunoprecipitation, visualizing the captured RNAs using whole gene microarrays, and identifying functionally relevant targets through genetic screens. Keywords: Association with RNA-binding protein
Project description:Circadian behaviors are regulated by intrinsic biological clocks consisting of central molecular oscillators and output pathways. Despite significant progress in elucidating the central timekeeping mechanisms, the molecular pathways coupling the circadian pacemaker to overt rhythmic behavior and physiology remain elusive. The Drosophila LARK RNA-binding protein is a candidate for such a coupling factor. Previous research indicates that LARK functions downstream of the clock to mediate behavioral outputs. To better understand the roles of LARK in the Drosophila circadian system, we sought to identify RNA molecules associated with LARK in vivo, using a novel strategy that involves capturing the RNA ligands by immunoprecipitation, visualizing the captured RNAs using whole gene microarrays, and identifying functionally relevant targets through genetic screens. Experiment Overall Design: LARK-containing ribonucleoprotein complexes (LARK-RNPs) were precipitated from lysates of hand-dissected pharate adult brains using an affinity-purified anti-LARK antibody (around 1000 brains were used per immunoprecipitation experiment). A portion of each lysate was saved prior to immunoprecipitations (IPs) in order to measure the relative abundance of transcripts in a total RNA sample. RNAs extracted from the LARK-RNP and total RNA samples were labeled and hybridized to Drosophila whole-genome gene microarrays; signal intensities for individual genes were compared between samples to identify those RNAs that were enriched by immunoprecipitation (relative to their abundances in total RNA). RNAs that were selectively enriched in the LARK-RNP samples were considered to be potential targets of the RNA-binding protein. Experiment Overall Design: Due to the difficulty to dissect large amount of fly brains, only two such immunoprecipitation experiments were performed, each generating an IP RNA sample and a total RNA (control) sample. The amount of RNAs obtained from IP is very small thus only one array is used for each sample - i.e. there are only biological replicates and no technical replicate.
Project description:The goal of this study is to identify, in the head of adult flies, mRNA species whose expresson level are altered by overexpression of the Drosophila RNA-binding protein LARK in CNS neurons. Keywords: genetic modification, gene experssion profile
Project description:The goal of this study is to identify, in the head of adult flies, mRNA species whose expresson level are altered by overexpression of the Drosophila RNA-binding protein LARK in CNS neurons. Experiment Overall Design: RNA samples from adult head of the LARK overexpression flies (elav-gal4; uas-lark/+) and control flies were compared. One total RNA sample was isolated from each genotype, of which three technical replicates (repeating the labeling and hybridization processes) were generated, respectively.
Project description:Steppe lands in Europe are critically affected by habitat loss and fragmentation, and hold over 50% of IUCN Red List bird species in Europe. Dupont's Lark is a threatened steppe-specialist passerine whose European geographic range is restricted to Spain, with less than 2000 pairs and an annual population decline of - 3.9%. Its strongly fragmented habitat leads to a metapopulation structure in the Iberian Peninsula that includes 24 populations and 100 subpopulations. We present an updated Population Viability Analysis based on the latest scientific knowledge regarding distribution, population trends, breeding biology and connectivity. Our results predict metapopulation extinction in 2-3 decades, through a centripetal contraction process from the periphery to the core. The probability of extinction in 20 years was 84.2%, which supports its relisting to Endangered in Spain following IUCN criteria. We carried out a sensitivity analysis showing that some parameters, especially productivity and survival of adults and juveniles, help to increase metapopulation viability. Simulation of management scenarios showed that habitat restoration in a subset of key subpopulations had a positive effect on the overall metapopulation persistence. Translocations of a limited number of individuals from source to recipient locations may help to rescue the most endangered subpopulations without reducing the global time to extinction of the metapopulation. In addition, we identified the most critical areas for action, where local populations of the species are prone to extinction. This work suggests that the viability of the Dupont's Lark metapopulation could be improved and its risk of extinction reduced if urgent and localized conservation measures are applied. In the short-term, habitat loss and fragmentation due to ploughing, reforestation and infrastructures implementation in Dupont's Lark habitat must be avoided. Habitat restoration and translocations could help to avoid imminent extinction of critical subpopulations. Restoration of extensive grazing is recommended as the most effective way to achieve the long-term conservation of Dupont's Lark in Spain.
Project description:Species that pass repeatedly through narrow population bottlenecks (<100 individuals) are likely to have lost a large proportion of their genetic variation. Having genotyped 92 Raso larks Alauda razae, a Critically Endangered single-island endemic whose world population in the Cape Verdes over the last 100 years has fluctuated between about 15 and 130 pairs, we found variation at 7 of 21 microsatellite loci that successfully amplified, the remaining loci being monomorphic. At 6 of the polymorphic loci variation was sex-linked, despite the fact that these microsatellites were not sex-linked in the other passerine birds where they were developed. Comparative analysis strongly suggests that material from several different autosomes has been recently transferred to the sex chromosomes in larks. Sex-linkage might plausibly allow some level of heterozygosity to be maintained, even in the face of persistently small population sizes.