Habitat matters – Strong genetic and epigenetic differentiation in Linum catharticum from dry and wet grasslands

Plant species differ in their ecological amplitude, with some species occurring in very different habitats under strongly differentiated environmental conditions. We were interested in to what extent the occurrence ofLinum catharticumin dry calcareous grasslands (Bromion) and wet litter meadows (Molinion), two habitats on opposing ends concerning, for example, moisture level, is reflected on the genetic and epigenetic level. Using AFLP (amplified fragment length polymorphisms) and MSAP (methylation sensitive amplification polymorphisms) analyses, we studied the genetic and epigenetic variation ofL. catharticumfrom calcareous grasslands and litter meadows. From each habitat, we included five study sites with 16 individuals per sampling location. We observed lower genetic than epigenetic diversity, but considerable differentiation among habitats, which was stronger on the genetic than the epigenetic level. Additionally, we observed a strong correlation of genetic and epigenetic distance, irrespective of geographic distance. The dataset included a large portion of fragments exclusively found in individuals from one or the other habitat. Some epigenetic fragments even occurred in different methylation states depending on the habitat. We conclude that environmental effects act on both the genetic and epigenetic level, producing the clear differentiation among plant individuals from calcareous grasslands and litter meadows. These results may also point into the direction of ecotype formation in this species

Genetic variation of litter meadow species reflects gene flow by hay transfer and mowing with agricultural machines

Litter meadows, historically established for litter production, are species-rich and diverse ecosystems. These meadows drastically declined during the last decades along with decreasing litter use in modern livestock housing. The aim of our study was to identify the drivers of genetic variation in litter meadow species. Therefore, we tested whether genetic diversity and differentiation depend on habitat age, landscape structure, habitat quality, and/or population size. We analysed 892 individuals ofAngelica sylvestris,Filipendula ulmaria, andSuccisa pratensisfrom 20 litter meadows across the Allgau in Baden-Wurttemberg (Germany) using AFLP analyses. All study species showed moderate levels of genetic diversity, while genetic differentiation among populations was low. Neither genetic diversity nor differentiation were clearly driven by habitat age. However, landscape structure, habitat quality as well as population size revealed different impacts on the genetic diversity of our study species. Past and present landscape structures shaped the genetic diversity patterns ofA. sylvestrisandF. ulmaria. The genetic diversity ofF. ulmariapopulations was, moreover, influenced by the local habitat quality.S. pratensispopulations seemed to be affected only by population size. All explanatory variables represent past as well as present gene flow patterns by anthropogenic land use. Therefore, we assume that genetic diversity and differentiation were shaped by both historical creation of litter meadows via hay transfer and present mowing with agricultural machines. These land use practices caused and still cause gene flow among populations in the declining habitats

Genetic Variation of Typical Plant Species in Hay Meadows: The Effect of Land Use History, Landscape Structure, and Habitat Quality

Global changes in land use are threatening the diversity of many ecosystems on both the intra- and interspecific levels. Among these ecosystems are the species-rich hay meadows, which have drastically declined in quality and quantity, due to land use intensification or abandonment in recent decades. The remaining genetic resources of their plant species must therefore be protected. To determine the driving forces impacting genetic variation in common hay meadow species (Dactylis glomerata, Heracleum sphondylium, and Trifolium pratense), we used data on the land use history, historic and present landscape structure and habitat quality. Our results showed average genetic diversity within the study sites, with low differentiation levels and a high gene flow among grasslands. Land use history, landscape structure and habitat quality were found to be related to the distribution of genetic diversity in the studied species, highlighting the complex forces acting in these ecosystems and showing the specific impact of litter accumulation on genetic diversity. Both historic and current environmental variables influence genetic diversity, demonstrating the importance of the land use history of a habitat. The most important group of variables impacting genetic variation in all three species was the landscape structure (e.g., distance to the nearest-located urban area or grassland). Also important was the influence of litter cover on genetic diversity in D. glomerata, which provides an interesting starting point for further research

Genetic controls of short- and long-term stomatal CO2 responses in Arabidopsis thaliana

Background and Aims The stomatal conductance (g(s)) of most plant species decreases in response to elevated atmospheric CO2 concentration. This response could have a significant impact on plant water use in a future climate. However, the regulation of the CO2 induced stomatal closure response is not fully understood. Moreover, the potential genetic links between short-term (within minutes to hours) and long-term (within weeks to months) responses of g(s) to increased atmospheric CO2 have not been explored. Methods We used Arabidopsis thaliana recombinant inbred lines originating from accessions Col-0 (strong CO2 response) and C24 (weak CO2 response) to study short- and long-term controls of g(s) Quantitative trait locus (QTL) mapping was used to identify loci controlling short- and long-term g(s) responses to elevated CO2 as well as other stomata-related traits. Key Results Short- and long-term stomatal responses to elevated CO2 were significantly correlated. Both short-and long-term responses were associated with a QTL, at the end of chromosome 2. The location of this QTL was confirmed using near-isogonic lines and it was fine-mapped to a 410-kb region. The QTL did not correspond to any known gene involved in stomatal closure and had no effect on the responsiveness to abscisic acid. Additionally, we identified numerous other loci associated with stomatal regulation. Conclusions We identified and confirmed the effect of a strong QTL corresponding to a yet unknown regulator of stomatal closure in response to elevated CO2 concentration. The correlation between short- and long-term stomatal CO2 responses and the genetic link between these traits highlight the importance of understanding guard cell CO2 signalling to predict and manipulate plant water use in a world with increasing atmospheric CO2 concentration. This study demonstrates the power of using natural variation to unravel the genetic regulation of complex traits.Peer reviewe

Socioeconomic Inequalities in Newborn Care During Facility and Home Deliveries: A Cross Sectional Analysis of Data from Demographic Surveillance Sites in Rural Bangladesh, India and Nepal

Background: In Bangladesh, India and Nepal, neonatal outcomes of poor infants are considerably worse than those of better-off infants. Understanding how these inequalities vary by country and place of delivery (home or facility) will allow targeting of interventions to those who need them most. We describe socio-economic inequalities in newborn care in rural areas of Bangladesh, Nepal and India for all deliveries and by place of delivery. Methods: We used data from surveillance sites in Bangladesh, India and from Makwanpur and Dhanusha districts in Nepal, covering periods from 2001 to 2011. We used literacy (ability to read a short text) as indicator of socioeconomic status. We developed a composite score of nine newborn care practices (score range 0–9 indicating infants received no newborn care to all nine newborn care practices). We modeled the effect of literacy and place of delivery on the newborn care score and on individual practices. Results: In all study sites (60,078 deliveries in total), use of facility delivery was higher among literate mothers. In all sites, inequalities in newborn care were observed: the difference in new born care between literate and illiterate ranged 0.35–0.80. The effect of literacy on the newborn care score reduced after adjusting for place of delivery (range score difference literate-illiterate: 0.21–0.43). Conclusion: Socioeconomic inequalities in facility care greatly contribute to inequalities in newborn care. Improving newborn care during home deliveries and improving access to facility care are a priority for addressing inequalities in newborn care and newborn mortality

Evacetrapib and Cardiovascular Outcomes in High-Risk Vascular Disease

BACKGROUND: The cholesteryl ester transfer protein inhibitor evacetrapib substantially raises the high-density lipoprotein (HDL) cholesterol level, reduces the low-density lipoprotein (LDL) cholesterol level, and enhances cellular cholesterol efflux capacity. We sought to determine the effect of evacetrapib on major adverse cardiovascular outcomes in patients with high-risk vascular disease. METHODS: In a multicenter, randomized, double-blind, placebo-controlled phase 3 trial, we enrolled 12,092 patients who had at least one of the following conditions: an acute coronary syndrome within the previous 30 to 365 days, cerebrovascular atherosclerotic disease, peripheral vascular arterial disease, or diabetes mellitus with coronary artery disease. Patients were randomly assigned to receive either evacetrapib at a dose of 130 mg or matching placebo, administered daily, in addition to standard medical therapy. The primary efficacy end point was the first occurrence of any component of the composite of death from cardiovascular causes, myocardial infarction, stroke, coronary revascularization, or hospitalization for unstable angina. RESULTS: At 3 months, a 31.1% decrease in the mean LDL cholesterol level was observed with evacetrapib versus a 6.0% increase with placebo, and a 133.2% increase in the mean HDL cholesterol level was seen with evacetrapib versus a 1.6% increase with placebo. After 1363 of the planned 1670 primary end-point events had occurred, the data and safety monitoring board recommended that the trial be terminated early because of a lack of efficacy. After a median of 26 months of evacetrapib or placebo, a primary end-point event occurred in 12.9% of the patients in the evacetrapib group and in 12.8% of those in the placebo group (hazard ratio, 1.01; 95% confidence interval, 0.91 to 1.11; P=0.91). CONCLUSIONS: Although the cholesteryl ester transfer protein inhibitor evacetrapib had favorable effects on established lipid biomarkers, treatment with evacetrapib did not result in a lower rate of cardiovascular events than placebo among patients with high-risk vascular disease. (Funded by Eli Lilly; ACCELERATE ClinicalTrials.gov number, NCT01687998 .)

Genetic and Epigenetic Variation of Typical Grassland Species – Habitat Specific Processes and Implications for Conservation

European semi-natural grasslands are among the most species rich habitats in the world. They were historically shaped by anthropogenic land use and developed their species and genetic variation alongside the agricultural practices of the time. The lasting effects of historic processes can be ob-served until today, however recent global developments are affecting this diversity in an unprece-dented intensity and frequency. The present study aims to investigate the effects of land use his-tory, landscape structure as well as vegetation and habitat characteristics on genetic and epigenetic variation of common grassland species from different grassland habitats. Chapter One introduces the scientific context the present thesis is placed in. Processes shaping biodiversity, the characteristics of the grassland ecosystems studied in this thesis and the history of grassland ecosystems in general is reviewed. Finally, grassland habitats are discussed in the context of biodiversity and conservation. In Chapter Two and Three the influence of land use history, landscape structure and habitat quality on the genetic variation of typical grassland species from oat-grass and litter meadows was investi-gated. We used Amplified Fragment Length Polymorphism (AFLP) analyses to estimate genetic diversity and differentiation among populations from ancient and recent grasslands. Chapter Two describes the genetic variation patterns of three typical grassland species (Dactylis glomerata L., s. I., Heracleum sphondylium L. & Trifolium pratense L.), studied in oat-grass mead-ows. Each species was influenced by different explanatory variables, but most interestingly current landscape structure and habitat quality, i.e. the amount of litter, influenced genetic diversity in this grassland habitat. Historic landscape structure and land use history were of minor interest in this comparably recently established grassland type. Similarly, Chapter Three deals with the genetic variation in litter meadow plant species (Angelica sylvestris L., Filipendula ulmaria (L.) MAXIM & Succisa pratensis MOENCH). Historic as well as recent gene flow patterns influenced genetic variation of the species in this ecosystem, highlighting the current importance of historic processes. However, the most important process, from a conserva-tion point of view, was the extensive gene flow produced by seed transfer via mowing machines. Extending the study of the intraspecific variation in grassland ecosystem to the epigenetic level, the genetic as well as epigenetic variation in Linum catharticum L. from different contrasting habitats is presented in Chapter Four. Combining AFLP and Methylation Sensitive Amplification Polymorphism (MSAP) analyses, large differences among populations from wet litter meadows and dry calcareous grasslands were observed, which could not be explained by geographic distance alone, but rather by the different local habitat conditions. This result highlights the impact of local environmental conditions on the genetic as well as the epigenetic level, which likely led to the development of different ecotypes. In Chapter Five the results of the three main chapters are reviewed in the context of nature con-servation. Historic as well as current gene flow processes were found to be important determinants of current genetic variation in common grassland species. Additionally, various local environmental factors contributed to the genetic as well as epigenetic variation patterns. In conclusion, genetic diversity in different grassland ecosystems and their respective species are influenced by different historic and present processes. Thus, conservation strategies should pay tribute to historic land use practices and stochasticity, while decreasing the impact of current pro-cesses of fragmentation and habitat loss, to increase gene flow among remnants of species-rich grasslands. Additionally, appropriate management will enhance the habitat quality, thus improving the establishment of new individuals, thereby increasing genetic variation. Different practical con-cepts, such as autochthonous seed material and genetic conservation areas can play an important role in preserving genetic variation for grassland species

Microplastics Effects on Reproduction and Body Length of the Soil-Dwelling Nematode Caenorhabditis elegans

Microplastics (MP) are pervasive in the environment. There is ample evidence of negative MP effects on biota in aquatic ecosystems, though little is known about MP effects in terrestrial ecosystems. Given numerous entry routes of MP into soils, soil organisms are likely to be exposed to MP. We compared potential toxicological effects of MP from (i) low-density polyethylene (LDPE) (mean diameter ± standard deviation: 57 ± 40 μm) and (ii) a blend of biodegradable polymers polylactide (PLA) and poly(butylene adipate-co-terephthalate) (PBAT) (40 ± 31 μm) on the reproduction and body length of the soil-dwelling bacterivorous nematode Caenorhabditis elegans. Feed suspensions without (control) or with MP (treatments) at concentrations of 1, 10, and 100 mg MP L–1 were prepared and nematodes were exposed to those suspensions on agar plates until completion of their reproductive phase (∼6 days). Using Nile red-stained PLA/PBAT MP particles and fluorescence microscopy, we demonstrated the ingestion of MP by C. elegans into pharynges and intestines. Under MP exposure, nematodes had fewer offspring (up to 22.9%) compared to nematodes in the control group. This decline was independent on the plastic type. We detected a tendency toward greater decreases in offspring at higher concentrations. Despite hints of negative effects on nematode body length under MP exposure, we could not derive a consistent pattern. We conclude that in MP-contaminated soils, the reproduction of nematodes, central actors in the soil food web, can be affected, with potentially negative implications for key soil functions, e.g., the regulation of soil biogeochemical cycles.Peer Reviewe