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The influence of mycorrhizal symbiosis on ecosystem processes depends on the mycorrhizal type and status of plants. Early research hypothesized that the proportion of arbuscular mycorrhizal (AM) species decreases and of ectomycorrhizal (ECM) and ericoid mycorrhizal (ERM) species increases along increasing elevations and latitudes. However, there is very scarce information about this pattern along elevation gradients. We aimed to test this hypothesis and to describe the trends in plant mycorrhizal status by examining the Pyrenean mountain range (from 400 to 3400 m asl). The distribution of plant mycorrhizal types: AM, ECM, ERM, and non-mycorrhizal (NM) and status (obligately, OM, or facultatively, FM mycorrhizal plants, FM) were identified based on the Pyrenean Floristic Atlas and analyzed for climatic and edaphic drivers. The proportion of AM plants decreased slightly with elevation, while ECM species peaked at 1000 m asl. The proportion of ERM and NM plant species rose with increasing elevation. The proportion of FM species increased, and OM species decreased with increasing elevation. The change of AM and ECM species, and OM and FM species, along the elevational gradient, corresponds broadly to changes along the latitudinal gradient, driven by a combination of climatic and edaphic factors. Differently, the elevational occurrence of NM plant species is mainly driven only by climatic factors (low temperature) and that of ERM species by only edaphic factors (low pH). Large-scale macroecological studies (≥ 50 km grid cell) well reflect the effects of climate on the distribution of plant mycorrhizal traits, but local data (≤ 1 km grid cell) are needed to understand the effects of soil conditions and land use.

The fungistatic activity of a lactic acid bacterium, which had been isolated from yellow pitahaya cultures, against fungi associated with basal rot (Fusarium oxysporum and Fusarium fujikuroi) was measured in the present study. Its activity was assessed in three fractions: fermented (S1), metabolic products (S2), and biomass (S3), using two fermentation substrates: Man Rogosa Sharpe agar (MRS) and potato dextrose agar (PDA). The bacterium was molecularly identified as Lactobacillus plantarum. S3 reduced F. fujikuroi growth by 100% over 48 h of fermentation, which occurred during the stationary phase of bacterial growth. The three fractions’ fungistatic activity against F. fujikuroi depended on the substrate employed. The fermentation kinetic parameters for L. plantarum indicated that its specific growth rate was 0.46 h–1, with 93.63% substrate consumption, 0.045 kg kg–1 cell yield, and 0.54 kg kg–1 product yield. The kinetic parameters calculated will allow for bacteria production scaling. These in-vitro trials reveal L. plantarum’s possible application as a biocontrol agent for diseases associated with Fusarium. However, further ex-vivo and in-vivo researches are required to demonstrate its behavior in crops.

El complejo respiratorio bovino (CRB) es la principal causa de enfermedad y muerte en el ganado de engorda en todo el mundo. Es un síndrome infeccioso multifactorial provocado por distintos virus y bacterias que disminuyen la eficiencia productiva y ocasionan pérdidas económicas. En México, el CRB se ha reportado en todas las regiones donde se engorda ganado; sin embargo, esos reportes carecen de información sobre la presencia del coronavirus respiratorio bovino (CVB), haciendo necesario contar con herramientas de diagnóstico confiables y precisas para detectar la presencia de CVB en el ganado que se engorda en México, para proponer las medidas sanitarias apropiadas para su manejo clínico. En este trabajo, se desarrolló una plataforma de diagnóstico molecular de PCR en tiempo real (rt-PCR) que amplifica un fragmento de la proteína S del CVB en muestras de exudado nasal. Al aplicar la plataforma rt-PCR para CVB en bovinos de engorda en aparente estado de salud y con signos de enfermedad respiratoria asociados a CRB se encontró que 19/50 (38 %) resultaron positivos, confirmando la presencia de ese virus en el ganado de la región. Los resultados de este trabajo significan el primer reporte sobre la presencia del CVB asociado al CRB en la región ganadera del noroeste de México y sienta las bases para futuras investigaciones sobre papel que juega este virus en la presentación de la patología del CRB en los sistemas de explotación de bovinos de engorda en nuestra región y el país.

The genus Fusarium produces mycotoxins that are metabolites of low molecular weight that affect the quality of crops, and even more importantly, they affect the health of humans and animals. Among those that cause damage to health are tricho-thecenes, fumonisins, and zearalenones. The objective of this study was to quantify the mycotoxins produced by species of the genus Fusarium from a population of isolates obtained from different crops and locations in Colombia. From 206 isolates, only 14 amplified to regions associated with mycotoxins de-oxynivalenol (DON) belonging to the group of trichothecenes, fumonisin (FUM) and zearalenone (ZEA) using PCR. Each isolate with the presence of mycotoxins was conserved in potato dextrose agar (PDA) medium. Eight days after seeding in corn kernel medium, the samples were processed to perform the quantitative analysis of DON, ZEA and FUM using an ELISA kit based on enzyme-linked immunosorbent assays. The results show that mycotoxins were present in the evaluated isolates and their levels were above the standards regulated by Mercosur and the European Union. The use of immunosorbent assays using the ELISA technique becomes a useful tool to detect and quantify mycotoxins of species of the genus Fusarium that affect different crops in Colombia. RESUMEN El género Fusarium produce micotoxinas que son metabolitos de bajo peso molecular que afectan la calidad de los cultivos y especialmente la salud de humanos y animales. Entre las que causan daños a la salud se encuentran los tricotecenos, las fumonisinas y las zearalenonas. El objetivo de este estudio fue cuantificar las micotoxinas producidas por especies del género Fusarium de una población de aislamientos obtenida de diferentes cultivos y localidades de Colombia. De 206 aislamientos, sólo 14 amplificaron regiones asociadas a las micotoxinas deoxinivalenol (DON), que pertenece al grupo de los tricotecenos, fumonisina (FUM) y zearalenona (ZEA) usando PCR. Cada uno de los aislamientos con presencia de micotoxinas se conservaron en medio papa dextrosa agar (PDA). Ocho días después de la siembra en medio de granos de maíz, las muestras se procesaron para realizar el análisis cuantitativo de DON, ZEA y FUM utilizando un kit de ELISA basado en ensayos de inmunoabsorción ligados a una enzima. Los resultados revelan la presencia de micotoxinas en los aislamientos evaluados cuyos niveles se encuentran por encima de los estándares regulados por Mercosur y la Unión Europea. El uso de ensayos de inmunoabsorción por medio de la técnica de ELISA se convierte en una herramienta útil para detectar y cuantificar micotoxinas de especies del género Fusarium que afectan diferentes cultivos en Colombia.

The Coat Protein I (COPI) complex is a seven-subunit coatomer complex consisting of the α, β, β', γ, δ, ε, and ζ proteins. In Arabidopsis thaliana, COPI is required for retrograde transport from the Golgi to the endoplasmic reticulum, Golgi maintenance, and cell plate formation. During compatible pollination, vesicle recruitment to the pollen contact point is required for pollen hydration and pollen tube penetration. Here, to identify other aspects of trafficking involved in the acceptance of compatible pollen by stigmatic papillae and to determine their roles in compatible pollination, we characterized knockout lines of several isoforms of the COPI complex, including α1-COP, γ-COP, and ε-COP. Specifically, we characterized pollen grain adherence, pollen tube penetration, and seed set in the mutants. Of the mutant lines examined, α1-cop had the most severe phenotypes, including altered compatible pollen grain adherence and tube germination and reduced seed set, whereas the other lines had milder phenotypes but visibly retarded compatible pollen acceptance. This is the first study demonstrating that COPI complex subunits are required for the acceptance of compatible pollen.

El objetivo de este estudio fue detectar la presencia de circovirus porcino tipo 2 (PCV2) en hatos porcinos de Baja California, México, empleando la técnica de reacción en cadena de la polimerasa (PCR). Se colectaron 97 muestras de sangre de cerdos procedentes de 26 granjas. El ADN se extrajo a partir de la capa leucoplaquetaria y las muestras fueron agrupadas en 19 grupos de cinco muestras y uno de dos muestras para el análisis de PCR. Se utilizaron cebadores específicos para amplificar un fragmento de 264 pares de bases del ORF2 de PCV2 utilizando como control positivo una vacuna comercial. El fragmento viral se detectó en el 15% (3/20) de los grupos analizados. Este es el primer reporte en el que se evidencia la presencia de circovirus porcino tipo 2 en Baja California., The aim of this study was to detect the presence of porcine circovirus type 2 (PCV2) in pig herds of Baja California, Mexico, using the polymerase chain reaction (PCR) technique. Blood samples (n=97) were collected from 97 pigs in 26 farms. The DNA was extracted from the buffy coat and the samples were grouped into 19 groups of five samples and one of two samples for PCR analysis. Specific primers were used to amplify a PCV2 ORF2 fragment of 264 base pairs and using a commercial vaccine as a positive control. The viral fragment was detected in 15% (3/20) of the groups. This is the first report in which the presence of porcine circovirus type 2 in Baja California is evidenced.

The hemolytic uremic syndrome associated with diarrhea, a consequence of Shiga toxin (Stx)-producing Escherichia coli infection, is a common cause of pediatric acute renal failure in Argentina. Stx type 2a (Stx2a) causes direct damage to renal cells and induces local inflammatory responses that involve secretion of inflammatory mediators and the recruitment of innate immune cells. γδ T cells constitute a subset of T lymphocytes, which act as early sensors of cellular stress and infection. They can exert cytotoxicity against infected and transformed cells, and produce cytokines and chemokines. In this study, we investigated the activation of human peripheral γδ T cells in response to the incubation with Stx2a-stimulated human glomerular endothelial cells (HGEC) or their conditioned medium, by analyzing in γδ T lymphocytes, the expression of CD69, CD107a, and perforin, and the production of TNF-α and IFN-γ. In addition, we evaluated by confocal microscopy the contact between γδ T cells and HGEC. This analysis showed an augmentation in cellular interactions in the presence of Stx2a-stimulated HGEC compared to untreated HGEC. Furthermore, we observed an increase in cytokine production and CD107a expression, together with a decrease in intracellular perforin when γδ T cells were incubated with Stx2a-treated HGEC or their conditioned medium. Interestingly, the blocking of TNF-α by Etanercept reversed the changes in the parameters measured in γδ T cells incubated with Stx2a-treated HGEC supernatants. Altogether, our results suggest that soluble factors released by Stx2a-stimulated HGEC modulate the activation of γδ T cells, being TNF-α a key player during this process.

20 pages, 5 figures, 4 tables, supporting information https://doi.org/10.1111/ddi.13382.-- Data availability statement: Microsatellite genotypes are available on Dryad (DOI https://doi.org/10.5061/dryad.xgxd254gw), Aim: How historical and contemporary eco-evolutionary processes shape the patterns of genetic diversity and differentiation across species’ distribution range remains an open question with strong conservation implications. Focusing on the orange stony coral, Astroides calycularis, we (a) characterized the pattern of neutral genetic diversity across the distribution range; (b) gave insights into the underlying processes; and (c) discussed conservation implications with emphasis on a national park located on a hotspot of genetic diversity. Location: South Mediterranean Sea and Zembra National Park. Methods: We combined new data from 12 microsatellites in 13 populations located in the Centre and in the Western Periphery of the distribution range with a published dataset including 16 populations from the Western and Eastern Peripheries. We analysed the relationship among parameters of genetic diversity (He, Ar(g)) and structure (population-specific FST) and two measures of geographic peripherality. We compared two estimators of pairwise genetic structure (GST, DEST) across the distribution range. The evolutionary and demographic history of the populations following the Last Glacial Maximum was reconstructed using approximate Bayesian computations and maximum-likelihood analyses. We inferred the contemporary connectivity among populations from Zembra National Park and with the neighbouring area of Cap Bon. Results: We demonstrate a decrease in genetic diversity and an increase in genetic differentiation from the Centre to the Eastern and Western Peripheries of the distribution range. Populations from Zembra show the highest genetic diversity reported in the species. We identified a spillover effect towards Cap Bon. Main conclusions: The patterns of genetic diversity and differentiation are most likely explained by “the postglacial range expansion hypothesis” rather than the “central–peripheral hypothesis.” Enforcement of conservation measures should be considered to protect this genetic diversity pattern, in particular when considering the low effective population size inferred at many sites, JBL was funded by a Postdoctoral Grant (SFRH/BPD/74400/2010) from the Portuguese Foundation for Science and Technology (FCT). This research was supported by national funds through FCT within the scope of UIDB/04423/2020 and UIDP/04423/2020 and by the MIMOSA project funded by the Foundation Prince Albert II Monaco. JG, DGG and PL acknowledge the funding of the Spanish government through the “Severo Ochoa Centre of Excellence” accreditation (CEX2019-000928-S). This work was partially supported by the European Union's Horizon 2020 research and innovation program under grant agreement SEP-210597628 (FutureMARES)

In many rural regions of Mexico, beekeeping is an important socioeconomic activity which is currently facing various health issues, being the main one the presence of infectious diseases. The aim o...

Shrub encroachment is considered one of the greatest threats in the conservation of pastures, especially in abandoned territories and territories with low stocking density. However, in the Navarra Pyrenees and Basque mountains, with a high stocking density maintained during the last decades, the expansion of argoma shrubland (Ulex gallii), shows that grazing is not enough for its control and requires evaluating measures of clearing. To evaluate the efficiency of the burning and the mechanical clearing (with two treatments: at ground level and 10 cm above ground) in the control of the shrub, these treatments were applied in different plots of encroached grasslands in two pastoral territories (Sierra de Aralar and Belate). After the burning and clearing, the evolution of the shrub was compared in the cleared plots closed to grazing and in their replicas open to it. The results showed differences between the different treatments in both territories that may be related to the initial coverage of the shrubland. In general, burning was less effective in controlling shrubland and allowed a greater resprout than mechanical clearing. No significant differences were found between the two types of mechanical clearing (at ground level and at 10 cm above ground). Finally, grazing caused a delay in the recovery of the shrub in all casesbut it was not able to eliminate the shrubs completely. From the field observations, the opinions of the farmers and the results of similar experiences, some measures for grassland management were collected and discussed to correct or complement the clearing treatments in order to increase their efficiency.

Non-chemical modified native corn (NCS), rice (NRS), wheat (NWS), and waxy corn (WCS) starch dispersions (5.0 g/100 g water) were heated (90 °C, 20 min) for producing gelatinized starch dispersions (GSD). GSD were centrifuged and the supernatant starch fraction (SF) was used for preparation of O/W emulsions (dispersed phase mass fraction of 0.15). The interfacial activity of the different SFs was assessed by dynamic interfacial tension measurements. The SF from NCS, NRS and NWS showed significantly higher and faster interfacial tension decaying rates than the WCS SF. All the SFs were able to form O/W emulsions. The volume-weighted mean droplet diameter (d4,3) of the fresh emulsions were significantly different ranging from ∼1.4 to 19 μm. The emulsions made with SFs from NCS, NRS and NWS showed larger d4,3 and faster creaming rates than that made with SF of WCS. The latter produced an emulsion with practically unchanged d4,3 and slowest creaming rate with storage time. FTIR, XRD, zeta-potential measurements, and adsorption time relaxation constants allowed postulating that emulsion stability was related to the formation of inclusion starch-lipid complexes at the oil-water interface. This study provides insights about the phenomena occurring on O/W emulsions interface due to interactions between cooked starch fractions from different botanical origin and lipids, and which might occur during food processing.

BackgroundHost and niche shifts are a source of genomic and phenotypic diversification as evidenced in parasitism. Exemplary is core metabolism reduction as parasites adapt to a particular host, while the accessory genome often maintains a high degree of diversification. However, selective pressures acting on the genome of organisms that have undergone lifestyle or host change have not been fully investigated.ResultsHere, we developed a comparative genomics approach to study underlying adaptive trends in oomycetes, a eukaryotic phylum with a broad range of economically important plant and animal parasitic lifestyles. Our analysis reveals converging evolution on biological processes for oomycetes that have similar lifestyle. Besides, we find that certain functions, in particular carbohydrate metabolism, transport, and signaling, are important for host and environmental adaption in oomycetes.DiscussionGiven the high correlation between lifestyle and genome properties in our oomycete dataset and the convergent evolution of fungal and oomycete genomes, we have developed a model that predicts plant pathogen lifestyles with high accuracy based on functional annotations. Understanding how genomes and selective pressures correlate with lifestyle may be crucial to identify new emerging diseases and pandemic threats.

Amyloids have proven to be a widespread phenomenon rather than an exception. Many proteins presenting the hallmarks of this characteristic beta sheet-rich folding have been described to date. Particularly common are functional amyloids that play an important role in the promotion of survival and pathogenicity in prokaryotes. Here, we describe important developments in amyloid protein research that relate to microbe-microbe and microbe-host interactions in the plant microbiome. Starting with biofilms, which are a broad strategy for bacterial persistence that is extremely important for plant colonization. Microbes rely on amyloid-based mechanisms to adhere and create a protective coating that shelters them from external stresses and promotes cooperation. Another strategy generally carried out by amyloids is the formation of hydrophobic surface layers. Known as hydrophobins, these proteins coat the aerial hyphae and spores of plant pathogenic fungi, as well as certain bacterial biofilms. They contribute to plant virulence through promoting dissemination and infectivity. Furthermore, antimicrobial activity is an interesting outcome of the amyloid structure that has potential application in medicine and agriculture. There are many known antimicrobial amyloids released by animals and plants; however, those produced by bacteria or fungi remain still largely unknown. Finally, we discuss amyloid proteins with a more indirect mode of action in their host interactions. These include virulence-promoting harpins, signaling transduction that functions through amyloid templating, and root nodule bacteria proteins that promote plant-microbe symbiosis. In summary, amyloids are an interesting paradigm for their many functional mechanisms linked to bacterial survival in plant-associated microbial communities.

The stability of ecological communities is critical for the stable provisioning of ecosystem services, such as food and forage production, carbon sequestration, and soil fertility. Greater biodiversity is expected to enhance stability across years by decreasing synchrony among species, but the drivers of stability in nature remain poorly resolved. Our analysis of time series from 79 datasets across the world showed that stability was associated more strongly with the degree of synchrony among dominant species than with species richness. The relatively weak influence of species richness is consistent with theory predicting that the effect of richness on stability weakens when synchrony is higher than expected under random fluctuations, which was the case in most communities. Land management, nutrient addition, and climate change treatments had relatively weak and varying effects on stability, modifying how species richness, synchrony, and stability interact. Our results demonstrate the prevalence of biotic drivers on ecosystem stability, with the potential for environmental drivers to alter the intricate relationship among richness, synchrony, and stability., We were supported by US NSF Grants DEB-8114302, DEB8811884, DEB-9411972, DEB-0080382, DEB-0620652, DEB-1234162, and DEB0618210; the Nutrient Network (https://nutnet.org/) experiment from NSF Research Coordination Network Grant NSF-DEB-1042132; the New Zealand National Vegetation Survey Databank; and Institute on the Environment Grant DG-0001-13. Data (Dataset 56, SI Appendix, Supplementary Text S4) owned by NERC Database Right/Copyright NERC. Further support was provided by the Jornada Basin Long-Term Ecological Research project, Cedar Creek Ecosystem Science Reserve, and the University of Minnesota. The Rothamsted Long-term Experiments National Capability is supported by UK Biotechnology and Biological Sciences Research Council Grant BBS/E/C/000J0300 and the Lawes Agricultural Trust. This research was funded by Czech Science Foundation Grant GACR16-15012S and Czech Academy of Sciences Grant RVO 67985939. E.V. was funded by 2017 Program for Attracting and Retaining Talent of Comunidad de Madrid Grant 2017-T2/AMB-5406.

Questions. Compensatory dynamics are described as one of the main mechanisms that increase community stability, e.g., where decreases of some species on a year‐to‐year basis are offset by an increase in others. Deviations from perfect synchrony between species (asynchrony) have therefore been advocated as an important mechanism underlying biodiversity effects on stability. However, it is unclear to what extent existing measures of synchrony actually capture the signal of year‐to‐year species fluctuations in the presence of long‐term directional trends in both species abundance and composition (species directional trends hereafter). Such directional trends may lead to a misinterpretation of indices commonly used to reflect year‐to‐year synchrony. Methods. An approach based on three‐term local quadrat variance (T3) which assesses population variability in a three‐year moving window, was used to overcome species directional trend effects. This “detrending” approach was applied to common indices of synchrony across a worldwide collection of 77 temporal plant community datasets comprising almost 7,800 individual plots sampled for at least six years. Plots included were either maintained under constant “control” conditions over time or were subjected to different management or disturbance treatments. Results. Accounting for directional trends increased the detection of year‐to‐year synchronous patterns in all synchrony indices considered. Specifically, synchrony values increased significantly in ~40% of the datasets with the T3 detrending approach while in ~10% synchrony decreased. For the 38 studies with both control and manipulated conditions, the increase in synchrony values was stronger for longer time series, particularly following experimental manipulation. Conclusions. Species’ long‐term directional trends can affect synchrony and stability measures potentially masking the ecological mechanism causing year‐to‐year fluctuations. As such, previous studies on community stability might have overemphasised the role of compensatory dynamics in real‐world ecosystems, and particularly in manipulative conditions, when not considering the possible overriding effects of long‐term directional trends., We thank multiple entities for the financial support necessary to obtain the different databases: the U.S. National Science Foundation under grant numbers DEB‐8114302, DEB‐8811884, DEB‐9411972, DEB‐0080382, DEB‐0620652, DEB‐1234162, DEB‐9707477, DEB‐0316402, DEB‐08‐16453, and DEB‐12‐56034, DEB‐0618210, the Nutrient Network (http://www.nutnet.org) experiment from the National Science Foundation Research Coordination Network (NSF‐DEB‐1042132), the New Zealand National Vegetation Survey Databank, the Spanish MINECO (Project CGL2014‐53789‐R), the Madrid Regional Government (Projects REMEDINAL‐3 and REMEDINAL‐TE), the European Research Council Synergy grant 610028 (IMBALANCE‐P), the Institute on the Environment (DG‐0001‐13), the SOERE‐ACBB financed through French National Agency for Research (ANAEE‐F, ANR‐11‐INBS‐0001), the Estonian Research Council (IUT 20‐28, IUT 20‐29), Czech Science Foundation (GAČR 17‐05506S and 19‐28491X), the European Regional Development Fund (Centre of Excellence EcolChange), the German Federal Environmental Foundation (DBU) for a grant to the NABU Hamburg (management experiment Calamagrostis epigejos), and the German Federal Ministry of Education and Research within the framework of the project BIOTA Southern Africa (promotion numbers 01LC0024, 01LC0024A and 01LC0624A2), Task 159 of SASSCAL (promotion number 01LG1201) and the Scottish Government's Rural and Environmental Science and Analytical Services division. Acknowledgement Data owned by NERC© Database Right/Copyright NERC. Further support was provided by the Jornada Basin Long‐Term Ecological Research (LTER) project, Cedar Creek Ecosystem Science Reserve and the University of Minnesota. We also thank the Lawes Agricultural Trust and Rothamsted Research for data from the e‐RA database. The Rothamsted Long‐term Experiments National Capability (LTE‐NCG) is supported by the UK Biotechnology and Biological Sciences Research Council (Grant BBS/E/C/000J0300) and the Lawes Agricultural Trust.

Resumen En el municipio de Ipiales, departamento de Nariño, se concentra la mayor área cultivada con arveja (Pisum sativum) de Colombia, con cerca de 3.070 ha, en donde se ha identificado un alto uso de plaguicidas asociado al establecimiento del modelo productivo. Por tanto, el objetivo de este estudio es evaluar el riesgo de impacto ambiental que genera la aplicación de los principales fungicidas, insecticidas, herbicidas y bactericidas. Para ello, se consideraron tres grupos de campesinos, previamente identificados como C1, C2 y C3 con una muestra de diez fincas por grupo. En una parcela de cada finca se determinó la materia orgánica, pH, textura, densidad aparente, distancia a fuentes hídricas y se caracterizaron los plaguicidas usados en dos ciclos de cultivo de arveja. La información recolectada fue ingresada al sistema de soporte IIRAmb para estimar el indicador I-PHY, el cual determinó el impacto ambiental de los plaguicidas. Los resultados sugieren la presencia de 44 ingredientes activos, con un uso promedio finca de 25,1 kg/ha/año; el indicador I-PHY global presentó diferencias estadísticas significativas entre C1 (4,04) respecto a C2 (5,06) y C3 (5,28), estos últimos grupos, mostraron el mayor riesgo ambiental principalmente por el tipo de moléculas usadas y las condiciones en las cuales se desarrolló el cultivo. A partir de estos resultados se genera una herramienta de monitoreo y evaluación, con el fin de mejorar las intervenciones en el sistema para incrementar el nivel de sustentabilidad. Abstract The Municipality of Ipiales (Department of Nariño, Colombia) concentrates the largest area cultivated with pea (Pisum sativum L.) in the country, nearly 3.070 ha, where an extensive use of pesticides has been associated with the implementation of a production-based model. Therefore, the objective of this study is to evaluate the environmental impact of the application of the most common fungicides, insecticides, herbicides, and bactericides. For that purpose, 30 farms were divided into three groups of ten farms each: C1, C2, and C3. The organic matter, pH, texture, apparent density, and distance to water sources of the plot of each farm were determined, and the pesticides used in the two cultivation cycles of pea were characterized. The collected information was entered into a support system, IIRAmb, in order to estimate the indicator, I-PHY, which determined the environmental impact of the pesticides. The results suggest the presence of 44 active ingredients, with an average use per farm of 25.1 kg/ha/year. The global I-PHY indicator presented significant statistical differences between C1 (4.04), C2 (5.06), and C3 (5.28). The last two groups showed the highest environmental risk, especially because the type of molecules they used and the conditions under which the crops developed. These results can be used to create a monitoring and evaluation tool in order to improve the adjustments to the system and increase its sustainability.

12 pages, 5 figures, 6 tables, supplemental material https://doi.org/10.25387/g3.12640691, The octocoral, Paramuricea clavata, is a habitat-forming anthozoan with a key ecological role in rocky benthic and biodiversity-rich communities in the Mediterranean and Eastern Atlantic. Shallow populations of P. clavata in the North-Western Mediterranean are severely affected by warming-induced mass mortality events (MMEs). These MMEs have differentially impacted individuals and populations of P. clavata (i.e., varied levels of tissue necrosis and mortality rates) over thousands of kilometers of coastal areas. The eco-evolutionary processes, including genetic factors, contributing to these differential responses remain to be characterized. Here, we sequenced a P. clavata individual with short and long read technologies, producing 169.98 Gb of Illumina paired-end and 3.55 Gb of Oxford Nanopore Technologies (ONT) reads. We obtained a de novo genome assembly accounting for 607 Mb in 64,145 scaffolds. The contig and scaffold N50s are 19.15 Kb and 23.92 Kb, respectively. Despite of the low contiguity of the assembly, its gene completeness is relatively high, including 75.8% complete and 9.4% fragmented genes out of the 978 metazoan genes contained in the metazoa_odb9 database. A total of 62,652 protein-coding genes have been annotated. This assembly is one of the few octocoral genomes currently available. This is undoubtedly a valuable resource for characterizing the genetic bases of the differential responses to thermal stress and for the identification of thermo-resistant individuals and populations. Overall, having the genome of P. clavata will facilitate studies of various aspects of its evolutionary ecology and elaboration of effective conservation plans such as active restoration to overcome the threats of global change, We acknowledge the funding support of the European Union’s Horizon 2020 research and innovation program under grant agreement No 689518 (MERCES) and the Strategic Funding UID/Multi/04423/2013 through national funds provided by FCT – Foundation for Science and Technology and European Regional Development Fund (ERDF), in the framework of the programme PT2020. JBL is funded by an assistant researcher contract framework of the RD Unit - UID/Multi/04423/2019 - Interdisciplinary Centre of Marine and Environmental Research – financed by the European Regional Development Fund (ERDF) through COMPETE2020 - Operational Program for Competitiveness and Internationalisation (POCI) and national funds through FCT/MCTES (PIDDAC). AA was partially supported by the FCT project PTDC/CTA-AMB/31774/2017 (POCI-01-0145-FEDER/031774/2017). DGG was supported by an FPU grant (FPU15/05457) from the Spanish Ministry of Education. We acknowledge support of the Spanish Ministry of Science and Innovation to the EMBL partnership, the Centro de Excelencia Severo Ochoa, the CERCA Programme / Generalitat de Catalunya, the Spanish Ministry of Science and Innovation through the Instituto de Salud Carlos III, the Generalitat de Catalunya through Departament de Salut and Departament d'Empresa i Coneixement, and the Co-financing by the Spanish Ministry of Science and Innovation with funds from the European Regional Development Fund (ERDF) corresponding to the 2014-2020 Smart Growth Operating Program. JBL and JG planned and granted the funding to start the project, With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Cell fitness and survival upon exposure to DNA damage depends on the repair of DNA lesions. Interestingly, cellular identity does affect and finetunes such response, although the molecular basis of such differences between tissues and cell types is not well understood. Thus, a possibility is that DNA repair itself is controlled by the mechanisms that govern cell identity. Here we show that the KLF4, involved in cellular homeostasis, proliferation, cell reprogramming and cancer development, directly regulates resection and homologous recombination proficiency. Indeed, resection efficiency follows KLF4 protein levels, i.e. decreases upon KLF4 downregulation and increases when is overexpressed. Moreover, KLF4 role in resection requires its methylation by the methyl-transferase PRMT5. Thus, PRMT5 depletion not only mimics KLF4 downregulation, but also showed an epistatic genetic relationship. Our data support a model in which the methylation of KLF4 by PRMT5 is a priming event required to license DNA resection and homologous recombination., This work was funded by R+D+I grants from the Spanish Ministry of Economy and Competitivity (SAF2013-43255-P, SAF2016-74855-P). FR-B is funded by the University of Sevillathrough a postdoctoral contract of the V PPIT-US. CABIMER is supported by the regional government of Andalucia (Junta de Andalucía).

The present work reports the discovery and the complete characterisation of an ancient cultivated rose variety found growing in a private garden in the southwest of the Principality of Asturias (northern Spain). The variety is here given the name Narcea. The majority of roses currently cultivated belong to the so-called group of ‘Modern Roses’, all of which were obtained after 1867 via artificial crosses and improvement programmes. All are destined for ornamental use. Until the 19th century, the great majority of the many ancient cultivated roses in Europe were used in perfumery and cosmetics, or had medicinal uses. Rosa damascena and Rosa centifollia are still grown and used by the French and Bulgarian perfume industries. The Asturian Massif of the Cantabrian Mountain Range provides a natural habitat for some 75% of the wild members of the genus Rosa, but until now there was no evidence that this area was home to ancient cultivated roses. A complete botanical description is here provided for a discovered ancient rose. It is also characterised according to a series of sequence tagged microsatellite sites, and its agronomic features are reported. In addition, a histological description (optical and scanning electronic microscope studies) of the petals is offered, along with an analysis of the volatile compounds present in these organs as determined by solid phase microextraction and gas chromatography-mass spectroscopy. The results reveal the uniqueness of this ancient type of rose and suggest it may be of interest to the perfume industry., Part of this work has been developed in the framework of the CSIC Interdisciplinary Thematic Platform ALCINDER.

Tree lines are supposed to react sensitively to the current global change. However, the lack of a long‐term (millennial) perspective on tree line shifts in the Pyrenees prevents understanding the underlying ecosystem dynamics and processes. We combine multiproxy palaeoecological analyses (fossil pollen, spores, conifer stomata, plant macrofossils, and ordination) from an outstanding ice cave deposit located in the alpine belt c. 200 m above current tree line (Armena‐A294 Ice Cave, 2,238 m a.s.l.), to assess for the first time in the Pyrenees, tree line dynamics, and ecosystem resilience to climate changes 5,700–2,200 (cal.) years ago. The tree line ecotone was located at the cave altitude from 5,700 to 4,650 cal year bp, when vegetation consisted of open Pinus uncinata Ramond ex DC and Betula spp. Woodlands and timberline were very close to the site. Subsequently, tree line slightly raised and timberline reached the ice cave altitude, exceeding its today's uppermost limit by c. 300–400 m during more than four centuries (4,650 and 4,200 cal year bp) at the end of the Holocene Thermal Maximum. After 4,200 cal year bp, alpine tundra communities dominated by Dryas octopetala L. expanded while tree line descended, most likely as a consequence of the Neoglacial cooling. Prehistoric livestock raising likely reinforced climate cooling impacts at 3,450–3,250 cal year bp. Finally, a tree line ecotone developed around the cave that was on its turn replaced by alpine communities during the past 2,000 years. Synthesis. The long‐term Pyrenean tree line ecotone sensitivity suggests that rising temperatures will trigger future P. uncinata and Betula expansions to higher elevations, replacing arctic–alpine plant species. Climate change is causing the rapid melting of the cave ice; rescue investigations would be urgently needed to exploit its unique ecological information.

The octocoral, Paramuricea clavata, is a habitat-forming anthozoan with a key ecological role in rocky benthic and biodiversity-rich communities in the Mediterranean and Eastern Atlantic. Shallow populations of P. clavata in the North-Western Mediterranean are severely affected by warming-induced mass mortality events (MMEs). These MMEs have differentially impacted individuals and populations of P. clavata (i.e. varied levels of tissue necrosis and mortality rates) over thousands of kilometers of coastal areas. The eco-evolutionary processes and genetic factors contributing to these differential responses remain to be characterized. Here, we sequenced a P. clavata individual with short and long read technologies, producing 169.98 Gb of Illumina paired-end and 3.55 Gb of Oxford Nanopore Technologies (ONT) reads. We obtained a de novo hybrid assembly accounting for 712.4 Mb and 107,682 scaffolds. The contig and scaffold N50 are 15.85 Kb and 17.01 Kb, respectively. Despite of the low contiguity of the assembly, the gene completeness was relatively high, including 86% of the 978 metazoan genes contained in the metazoa_odb9 database. A total of 76,508 protein-coding genes and 85,763 transcripts have been annotated. This assembly is one of the few octocoral genomes currently available. This is undoubtedly a valuable resource for characterizing the genetic bases of the differential responses to thermal stress and for the identification of thermo-resistant individuals and populations. Overall, the genome of P. clavata will help to understand various aspects of its evolutionary ecology and to elaborate effective conservation plans such as active restoration actions to overcome the threats of global change.

Summary Acquired genomic instability is one of the major concerns for the clinical use of induced pluripotent stem cells (iPSCs). All reprogramming methods are accompanied by the induction of DNA damage, of which double-strand breaks are the most cytotoxic and mutagenic. Consequently, DNA repair genes seem to be relevant for accurate reprogramming to minimize the impact of such DNA damage. Here, we reveal that reprogramming is associated with high levels of DNA end resection, a critical step in homologous recombination. Moreover, the resection factor CtIP is essential for cell reprogramming and establishment of iPSCs, probably to repair reprogramming-induced DNA damage. Our data reveal a new role for DNA end resection in maintaining genomic stability during cell reprogramming, allowing DNA repair fidelity to be retained in both human and mouse iPSCs. Moreover, we demonstrate that reprogramming in a resection-defective environment has long-term consequences on stem cell self-renewal and differentiation., Graphical Abstract, Highlights • DNA end resection mechanism is hyper-activated during reprogramming • CtIP protein levels increase during mouse and human cell reprogramming • Loss of CtIP impairs mouse and human reprogramming efficiency • Reprogramming without CtIP causes defects in iPSC self-renewal and differentiation, Therapeutic use of iPSC is limited due to the genomic alterations that accompany cell reprograming. Huertas, Gómez-Cabello, and colleagues show that DNA resection is essential to avoid such phenomena and that CtIP is critically required for mouse and human cell reprogramming. Indeed, reprogramming in the absence of CtIP has severe consequences for cell maintenance and differentiation.

Aim To assess the spatial congruence between hotspots based on taxonomic, phylogenetic and functional diversity, after accounting for the correlation between diversity metrics, and the spatial scale and sampling completeness of data.Location The Ordesa and Monte Perdido National Park (Central Pyrenees, Spain), a species-rich area subjected to intensive botanical sampling.Methods We selected hotspots using different diversity metrics and two different data sources (similar to 49,000 occurrence records of 1379 vascular plants in 1 9 1 km grid cells and 1218 inventories of plant communities containing a total of 859 taxa) and compared their spatial congruence. The effect of sampling completeness of data was explicitly assessed. Phylogenetic diversity and functional diversity (measured with richness-dependent and richness-independent metrics) were based on a molecular phylogeny, and a functional dendrogram, respectively. The effectiveness of different types of hotspots in representing other diversity components was tested with permutation tests.Results We found that spurious correlations between diversity metrics explained the congruence between taxonomic, phylogenetic and functional hotspots. When richness-independent metrics were used, diversity hotspots were no longer congruent regardless of the source of data. Hotspots were biased towards intensively sampled grid cells, and the amount of diversity they captured was exaggerated due to the coarse spatial scale of species-occurrence data. The efficiency of hotspots in terms of integrating different diversity components was lower at community scale and not significantly higher than expected at random, regardless of the sampling completeness.Main conclusions Our results stress that the arbitrary use of diversity metrics and the scale of analyses along with the sampling bias in data can distort the true location of hotspots, and exaggerate their spatial congruence. After accounting for such methodological issues, we found a clear mismatch between diversity components that questions the utility of hotspots as a conservation tool of multiple diversity components.

Este trabajo ha sido financiado con fondos de los proyectos CICYT Ministerio de Educación y Ciencia (CGL2007−60533/BOS, CGL2010−18312/BOS) y del Ministerio de Economía y Competitividad (CGL2014−53308−P), de la red REMEDINAL (S−0505/AMB/ 0335, S2009 AMB−1783, S2013/MAE−2719 y TE-CM S2018/EMT- 4338), de la Universidad de Alcalá (CCG2014/BIO−002) y del MAGRAMA y la Fundación Biodiversidad a través de la Fundación Internacional para la Restauración de Ecosistemas. GGS fue financiado con una beca FPU (AP2006−00891) y SMH por una beca FPI (BES−2008−006630), ambas del Ministerio de Educación y Ciencia. LMB fue financiada por una beca FPI (BES-2015-075276) del Ministerio de Ciencia y Tecnología. NM fue financiada por una beca de la Universidad de Alcalá. ABL agradece ayudas postdoctorales Juan de la Cierva (FJCI-2015-23687) y de la Fundación Alexander von Humboldt. Agradecemos la colaboración de la Dirección Xeral de Conservación da Natureza de la Xunta de Galicia por las facilidades para desarrollar las investigaciones realizadas en esta Comunidad y de las bodegas de Abadía Retuerta y Martín Berdugo y de Haciendas Bio. Finalmente, agradecemos la contribución de un revisor anónimo que mejoró la versión original de este artículo., Los elementos que soportan las funciones y servicios de los ecosistemas son insuficientemente conocidos, mermando nuestra capacidad para planificar la conservación, restauración y manejo de los ecosistemas. Entre dichos elementos, el papel de la avifauna es especialmente relevante al ser especies de gran movilidad (high mobile link species) que conectan distintas unidades del paisaje. Este trabajo analiza servicios ecosistémicos importantes dependientes de interacciones biológicas mediadas por la avifauna en mosaicos agroforestales. Abordamos tres objetivos específicos: el papel de (1) los córvidos en la dispersión y reclutamiento de quercíneas, (2) la avifauna insectívora en la regulación de plagas en cultivos leñosos y (3) las aves rapaces en la regulación de avifauna-plaga en viñedos. Nuestros resultados evidencian que la avifauna realiza funciones y servicios claves para la regeneración forestal y la regulación de plagas en mosaicos agroforestales. Identificamos especies que se desconocía que realizaban una función relevante, en particular la urraca (Pica pica), cuya actividad resulta en una dispersión efectiva de nueces y bellotas, y el abejero europeo (Pernis apivorus), con un elevado consumo de avispa asiática (Vespa velutina). La instalación de cajas-nido en cultivos leñosos aumenta las poblaciones de aves insectívoras que regulan plagas; sin embargo, la capacidad de regulación de las plagas es moderada y depende del contexto ambiental o estudio de caso. Las rapaces diurnas existentes en los paisajes estudiados contribuyen al control de plagas agrícolas como la del estornino negro (Sturnus unicolor). Proponemos integrar los servicios que proporcionan las aves en la ordenación del territorio para alcanzar modelos de desarrollo más sostenibles., The elements that support functions and ecosystem services are not well known yet, and this gap reduces our capacity to plan the conservation, restoration, and management of ecosystems. Among these elements, the role of avifauna is especially relevant as birds are high-mobile link species that connect different landscape units. This article analyzes important ecosystem services driven by biological interactions that are mediated by birds in farmland and forest mosaics. We propose three specific goals: to assess the role of (1) corvids in the dispersal and recruitment of oak species, (2) insectivorous birds in pest regulation of woody crops, and (3) raptors in regulation of plague birds in vineyards. Our results show that the avifauna performs key functions and services for forest regeneration and pest regulation in agroforestry mosaics. We identified species that were not known to perform relevant functions, particularly the magpie (Pica pica) and its effective dispersion of nuts and acorns, and the honey buzzard (Pernis apivorus) and its high consumption of Asian black hornet (Vespa velutina). The installation of nest-boxes in woody crops increases the populations of insectivorous birds that regulate pests; however, the capacity of pest control is moderate and depends on the environmental context or case study. The diurnal raptors thriving in the studied landscapes contribute to the control of agricultural pests such as the spotless starling (Sturnus unicolor). We propose to integrate the services provided by birds in landscape planning to achieve more sustainable development systems., Financiado con fondos de los proyectos CICYT Ministerio de Educación y Ciencia (CGL2007−60533/BOS, CGL2010−18312/BOS) y del Ministerio de Economía y Competitividad (CGL2014−53308−P), de la red REMEDINAL (S−0505/AMB/ 0335, S2009 AMB−1783, S2013/MAE−2719 y TE-CM S2018/EMT- 4338), de la Universidad de Alcalá (CCG2014/BIO−002) y del MAGRAMA y la Fundación Biodiversidad a través de la Fundación Internacional para la Restauración de Ecosistemas. GGS fue financiado con una beca FPU (AP2006−00891) y SMH por una beca FPI (BES−2008−006630), ambas del Ministerio de Educación y Ciencia. LMB fue financiada por una beca FPI (BES-2015-075276) del Ministerio de Ciencia y Tecnología. NM fue financiada por una beca de la Universidad de Alcalá. ABL agradece ayudas postdoctorales Juan de la Cierva (FJCI-2015-23687) y de la Fundación Alexander von Humboldt

The vicuna (Vicugna vicugna) is the most representative wild ungulate of the high Andes of South America with two recognized morphological subspecies, V. v. mensalis in the north and V. v. vicugna in the south of its distribution. Current vicuna population size (460,000–520,000 animals) is the result of population recovery programs established in response to 500 years of overexploitation. Despite the vicuna’s ecosystemic, economic and social importance, studies about their genetic variation and history are limited and geographically restricted. Here, we present a comprehensive assessment of the genetic diversity of vicuna based on samples collected throughout its distribution range corresponding to eleven localities in Peru and five in Chile representing V. v. mensalis, plus four localities each in Argentina and Chile representing V. v. vicugna. Analysis of mitochondrial DNA and microsatellite markers show contrasting results regarding differentiation between the two vicuna types with mitochondrial haplotypes supporting subspecies differentiation, albeit with only a few mutational steps separating the two subspecies. In contrast, microsatellite markers show that vicuna genetic variation is best explained as an isolation by distance pattern where populations on opposite ends of the distribution present different allelic compositions, but the intermediate populations present a variety of alleles shared by both extreme forms. Demographic characterization of the species evidenced a simultaneous and strong reduction in the effective population size in all localities supporting the existence of a unique, large ancestral population (effective size ∼50,000 individuals) as recently as the mid-Holocene. Furthermore, the genetic variation observed across all localities is better explained by a model of gene flow interconnecting them rather than only by genetic drift. Consequently, we propose space “continuous” Management Units for vicuna as populations exhibit differentiation by distance and spatial autocorrelation linked to sex biased dispersal instead of population fragmentation or geographical barriers across the distribution.

Approximately half of the area in the Spanish Central Pyrenees is dedicated to pastures. A decrease in stocking rate coupled with changes in livestock management in recent decades have favoured the expansion of Nardus grasslands, which are considered undesirable for grazing use and for diversity conservation. The objective of this study was to analyse how topsoil properties are related to grassland plant composition occurring in erosion-disturbed (chalk grasslands) and undisturbed (Nardus mat-grasslands) soils in a subalpine area of the Spanish Central Pyrenees. We selected six paired sampling points for a side-by-side comparison of both communities. At each point, we 1) estimated the plant cover of each species through inventories and 2) analysed a set of physical-chemical topsoil properties (0-5 and 5-10 cm depth). Data were analysed through multivariate analysis. We found typical species of Nardus mat-grasslands in the undisturbed sites growing on non-eroded and well-structured soils that were low in calcium and acidic, with high contents of organic matter. In turn, we found earlier-successional grassland communities growing on slopes recently affected by soil erosion processes. The species composition was mainly species from stony slope grasslands and, to a lesser extent, from the long-term snow-covered environments of the high mountains. These soils were shallower and stonier and had a less-stable structure, higher pH, and lower organic matter and calcium content than undisturbed soils. Our results suggest that the differences between both communities emerge and are maintained by soil-plant feedback mechanisms mediated in Nardus mat-grasslands through soil stabilization and acidification and in chalk grasslands through soil erosion and basification. These findings suggest that the subalpine grassland mosaic results from a model of non-equilibrium plant coexistence due to soil disturbance and inexorable succession. Management should be focused on maintaining a disturbance regime, through grazing, sufficient to prevent the spreading of Nardus mat-grasslands.

In the present scenario of climatic change, climatic refugia will be of paramount importance for species per- sistence. Topography can generate a considerable climatic heterogeneity over short distances, which is often disregarded in macroclimatic predictive models. Here we investigate the role of rocky habitats as microclimatic refugia by combining two different analyses: exploring a thermal mechanism whereby rocky habitats might serve as refugia, and examining if the biogeographic pattern shows a high abundance of relict, endemic and peripheral species. The thermal profile of two populations of relict and endemic plant species occurring in Pyrenean cliffs was investigated by infrared images and in situ temperature data-loggers. Despite occurring in crevices of a south oriented slope, Androsace cylindrica showed a narrower daily range of temperature than the surrounding matrix, thereby avoiding extreme high temperatures. Borderea chouardii, of tropical ancestors, also occurred in patches where temperatures were buffered during the growth season, experiencing lower mean temperatures than the surrounding matrix and nearby areas during the warmer part of the day, and similar temperatures during the colder. The rocky habitats of both species, therefore, reduced temperature ranges and exposition to extreme climatic events. Compared to other habitats, the rocky ones also harboured a very high fraction of both endemics and peripheral plant populations according to the largest vegetation dataset available in the Pyrenees (18,800 plant inventories and 400,000 records). Our results suggest an association between the habitats of relicts, en- demics and species at their distribution limit, driven by a stabilizing effect of rocky habitats on extreme tem- peratures. Given the important role of rocky habitats as hotspots of singular and unique plants, their char- acterization seems a sensible first step to identify potential refugia in the context of climate change.

Leptospira infects a wide range of companion, domestic and wild animal species, shedding the spirochetes into the environment via urine. Dogs become infected by direct or indirect contact with wild or domestic infected animal reservoirs increasing the risk of zoonotic transmission of the disease. The microscopic agglutination test has been used as the gold standard for the diagnosis of leptospirosis but has low sensitivity and is technically complex. Several ELISA tests have been developed based on recombinant proteins of Leptospira for the diagnosis of leptospirosis with similar or higher specificity and sensitivity levels than the microscopic agglutination test. Here, we developed and analytically validated an ELISA test based on recombinant LipL32 protein of Leptospira expressed in baculovirus. The LipL32 protein was successfully adapted in an indirect ELISA using dog plasma samples. Optimization of the ELISA resulted in a P/N ratio of 7.18 using only 5 ng of rLipL32 per well. Inter-assay and intra-assay variation showed a CV of 3.96% and 6.98% respectively, suggesting that the ELISA-LipL32 is highly reproducible. When tested with field samples, concordance of the ELISA-LipL32 with a real-time PCR, positive concordance was 100%. Our results indicate that the ELISA-LipL32 has the potential to be used by veterinarians and public health investigators as a safe, rapid, inexpensive and reliable method for the early diagnosis of Leptospira infection in dogs. Additional studies are still required for clinical validation on field samples under different epidemiological scenarios.

RAD51 paralogs are a group of conserved proteins in eukaryotes that are involved in the repair of DNA breaks at several levels. On one hand, they help the strand invasion step catalyzed by RAD51. Also, they play late roles in Holliday Junction metabolism. Here we uncover a new role of the RAD51 paralogs at an earlier event in the repair of broken chromosomes. All five RAD51 paralogs affect the balance between double strand break repair pathways. Specifically, they favor homology-mediated repair over non-homologous end-joining. Such role is independent of RAD51 or the checkpoint activity of these proteins. Moreover, it defines a novel control point of double strand break repair independent and subsequent to DNA-end resection initiation.Mechanistically, RAD51 paralogs limit the retention of Ku80 at the sites of DNA breaks. Thus, our data extend the role of this family of proteins to the earliest event of double strand break repair.

Understanding the effects of herbivores on plant communities is needed for anticipating how variation in grazing regimes will impact natural and semi-natural ecosystems. Prominent ecological hypotheses predict that drastic reductions of herbivory, as have occurred in many European mountains, will trigger fast diversity loss and structural changes in grasslands, because grazing tolerant species are expected to be rapidly replaced by taller and more competitive species. The aims of this study were to test generalizations of herbivory effects (mostly from lowlands) in high elevation grasslands over ecological relevant times scales, and to unravel the contribution of climatic conditions to observed changes. Species richness, canopy height, plant community structure, and transition probabilities among ecological groups were monitored throughout 19 years in herbivore exclusions and control plots in the Central Pyrenees. We used ordination analyses to track the long-term community response to herbivore exclusion, and generalized additive models to assess the non-linear effects of herbivore exclusion and climatic conditions on measured variables. Contrary to expectations, herbivore exclusion did not significantly affect species richness, and although canopy height increased, it was not drastic enough to suppress shade intolerant species. The strongest shifts in plant community structure and transition probabilities between ecological groups occurred during a sequence of warm and dry growing seasons, whereas in control plots, these changes were smaller, and largely reversed after cooler and wetter climatic conditions returned. Our results suggest that long-term effects of grazing cessation in high elevation grasslands can be weaker and slower than predicted. However, these effects can act synergistically with dry and warm events. Therefore, the maintenance of past grazing activities can be key in the face of ongoing climatic warming., We thank F. Fillat, J. Revilla and M. Gartzia for their technical assistance in the field, and M.M. Pata for statistical advice. We are grateful to the staff of the Ordesa-Monte Perdido National Park for the facilities provided through years. This manuscript benefited from the comments of various anonymous reviewers. This study was supported by the Spanish Ministry of Science and Technology through a doctoral grant to I.P. (AP2008-01449), and two projects of National Parks (018/2008 and 430/211; http://​www.​idi.​mineco.​gob.​es/​portal/​site/​MICINN/​).

Sequencing whole genomes has become a standard research tool in many disciplines including Molecular Ecology, but the rapid technological advances in combination with several competing platforms have resulted in a confusing diversity of formats. This lack of standard formats causes several problems, such as undocumented preprocessing steps or the loss of information in downstream software tools, which do not account for the specifics of the different available formats. ReadTools is an open-source Java toolkit designed to standardize and preprocess read data from different platforms. It manages FASTQ- and SAM-formatted inputs while dealing with platform-specific peculiarities and provides a standard SAM compliant output. The code and executable are available at https://github.com/magicDGS/ReadTools.

The Southern Pacific Coast of Costa Rica shows some local variations in its four Morpho species. The goal of this article is to compare the life cycle of M. amathonte from different areas in Costa Rica and South America. The immature stages were found on Pterocarpus officinalis Jacq. and then photographed and described so as to illustrate its morphology and behavior. It is clear, that M. amathonte from the South Pacific side of Costa Rica comes from one isolated population and demonstrates a cryptic nature between the two forms present in Costa Rica., Las cuatro especies de Morpho de la costa Pacífica de Costa Rica muestra algunas variaciones locales. El objetivo de este artículo es comparar el ciclo de vida de M. amathonte de diferentes localidades de Costa Rica y Sudamérica. Los estadios inmaduros se encontraron en Pterocarpus officinalis Jacq. y luego fotografiados y descritos para ilustrar su morfología y comportamiento. Es evidente que M. amathonte del lado del Pacífico de Costa Rica proviene de una población aislada y demuestra una naturaleza críptica entre las dos formas presentes en Costa Rica.

Zoonotic parasitic infections are a major global public and veterinary health problem and widespread among straydogs. The objective of this study was to establish the prevalence of intestinal parasites in stray dogs in the urban, rural and coastalareas of Mexicali County in northwest Mexico. In 2014, from January to December, 380 stray dogs were captured. The entire smallintestine, cecum and faeces samples were collected and examined by using simple zinc sulfate flotation and Lugol’s solution staining.Data were statistically analysed. Overall, about 21.5% of examined dogs were found positive for intestinal parasites. Toxocara caniswas the most frequent detected parasite, with a prevalence of 7.1%, followed by Toxascaris leonina (5.5%), Cystoisospora spp.(5.0%), Taenia spp. (3.9%) and Dipylidium caninum (2.8%). Dogs were more frequently found to be infected with a single genus of intestinal parasite (18.7%) than co-infected (2.8%). Intestinal parasites were more prevalent in samples from the coastal area (25%)than in those from the rural (24.4%) and urban (20.6%) areas, however, only statistical association was found between capture areaand specific intestinal parasitic infection. There were significant differences in the prevalence of taeniasis among two age groups (P, RESUMEN. Las infecciones zoonóticas parasitarias son un problema global público y para la medicina veterinaria, siendo diseminadas por perros callejeros. El objetivo del estudio fue determinar la prevalencia de parasitosis intestinales en perros callejerosde la zona urbana, rural y costera del municipio de Mexicali, noroeste de México. En 2014, entre enero y diciembre, se capturaron380 perros. Se colectó el intestino, ciego y heces y se examinaron utilizando flotación con sulfato de zinc y tinción con la soluciónde Lugol. Los datos fueron analizados estadísticamente. En general, alrededor del 21,5% de los perros examinados fueron positivosa parásitos intestinales. Toxocara canis fue el parásito más frecuentemente, con una prevalencia del 7,1%, seguido por Toxascarisleonina (5,5%), Cystoisospora spp. (5,0%), Taenia spp. (3,9%) y Dipylidium caninum (2,8%). Los perros fueron más frecuentementeencontrados infectados con un solo género de parásito intestinal (18,7%) que coinfectados (2,8%). Las parasitosis intestinales fueronmás prevalentes en muestras de la costa (25%) que del área rural (24,4%) y urbana (20,6%), sin embargo solo se encontró asociaciónestadística entre el área de captura y las parasitosis intestinales específicas. Hubo diferencias significativas en la prevalencia de taeniasisentre los dos grupos de edad (P

There are two major and alternative pathways to repair DNA double-strand breaks: non-homologous end-joining and homologous recombination. Here we identify and characterize novel factors involved in choosing between these pathways; in this study we took advantage of the SeeSaw Reporter, in which the repair of double-strand breaks by homology-independent or -dependent mechanisms is distinguished by the accumulation of green or red fluorescence, respectively. Using a genome-wide human esiRNA (endoribonuclease-prepared siRNA) library, we isolate genes that control the recombination/end-joining ratio. Here we report that two distinct sets of genes are involved in the control of the balance between NHEJ and HR: those that are required to facilitate recombination and those that favour NHEJ. This last category includes CCAR2/DBC1, which we show inhibits recombination by limiting the initiation and the extent of DNA end resection, thereby acting as an antagonist of CtIP., This work was funded by a R + D + I grant from the Spanish Ministry of Economy and Competitivity (SAF2010-14877) and an ERC Starting Grant (DSBRECA). A.L.-S. is the recipient of a FPI fellowship from the Spanish Ministry of Economy and Competitivity, and F.M.-N. is funded with an FPU fellowship from the Spanish Ministry of Education. M.I.M.-M. was the recipient of a Juan de la Cierva Postdoctoral Grant. M.S.D.-S. received a EMBO short-term fellowship for this project. J.B. and C.D. were supported by grants from the Danish National Research Foundation (CARD; DNRF125), the Danish Council for Independent Research (DFF-1331-00262) and the Novo Nordisk Foundation (NNF16584).

Marinomonas mediterranea is a marine gamma-proteobacterium that synthesizes LodA, a novel L-lysine-ε-oxidase (E.C. 1.4.3.20). This enzyme oxidizes L-lysine generating 2-aminoadipate 6-semialdehyde, ammonium, and hydrogen peroxide. Unlike other L-amino acid oxidases, LodA is not a flavoprotein but contains a quinone cofactor. LodA is encoded by an operon with two genes, lodA and lodB. In the native system, LodB is required for the synthesis of a functional LodA. In this study, we report the recombinant expression of LodA in Escherichia coli using vectors that allow its expression and accumulation in the cytoplasm. To reveal the L-lysine-ε-oxidase activity using the Amplex Red method for hydrogen peroxide detection, it is necessary to first remove the E. coli cytoplasmic catalases. The flavoprotein LodB is the only M. mediterranea protein required in the recombinant system for the generation of the cofactor of LodA. In the absence of LodB, LodA does not contain the quinone cofactor and remains in an inactive form. The results presented indicate that LodB participates in the posttranslational modification of LodA that generates the quinone cofactor.

En el Magdalena Medio colombiano confluyen dos actividades económicas: una ganadería intensiva y una consolidada industria petroquímica. Esta última representa un potencial peligro para la Salud Pública, como consecuencia de la emisión de metales tóxicos (plomo y cadmio) que pueden incorporarse a la cadena trófica. Por ello se realizó un inventario de pasturas en esta región en función de la distancia (100, 500, 2500 y 5000 m) al foco emisor contaminante. Así se muestrearon en zonas próximas a la refinería de crudo (Barrancabermeja, Santander), en pozos de extracción (Yondó, Antioquía) y tomando como testigo el Campus académico de la Universidad de la Paz (Barrancabermeja, Santander). Los resultados obtenidos indican una alta frecuencia de representantes de la familia Poaceae y de sus géneros de Brachiaria spp., con importantes diferencias. La información se interpretó en los tres primeros componentes, donde las variables con mayor peso fueron la profundidad del suelo a 5 y 30 cm, tipo de pastura con predominio de las especies introducidas y el transecto en relación a la distancia focal de las fuentes de contaminación. Así, en la zona de la refinería solo se han detectado dos especies de Brachiaria, mientras que en la zona de los pozos de extracción y en la zona testigo se presenta mayor frecuencia de otras especies de Brachiaria. Se concluye que los mayores niveles de bioacumulación de metales en tejidos vegetales presentó el siguiente orden: raíz > tallo > hojas, a su vez la acumulación en suelo mostró ser mayor en la profundidad de 5 cm. In the Middle Magdalena Colombian converge two economic activities: intensive livestock and a consolidated petrochemical industry. The latter represents a potential danger to public health, resulting from the emission of toxic metals (lead and cadmium) that can enter in the food chain. Therefore pasture inventory is performed in this region depending on the distance (100, 500, 2500 and 5000 m) to the emitting source contaminant. So they were sampled near the oil refinery (Barrancabermeja, Santander) in extraction wells (Yondó, Antioquia) and sampling as witness the academic campus of the Universidad de la Paz (Barrancabermeja, Santander) areas. The results indicate a high frequency of representatives of the Poaceae family and its genera Brachiaria, with important differences. The information was interpreted in the first three components, where the variables with more weight were soil depth to 5 and 30 centimeters, type of pasture with predominance of introduced species and transect relative to the focal length of the sources of pollution. Then, in the area of the refinery were detected only two species of Brachiaria, while in the area of the extraction wells and in the control area it is presented a higher frequency of other species of Brachiaria. It_s concluded that higher levels of bioaccumulation of metals in plant tissues presented the following order: root > stem > leaves, turn the accumulation in soil was found to be higher in the depth of 5 cm.

ResumenEl objetivo de este trabajo fue identificar aislamientos patogénicos de Fusarium en frutos de pitahaya amarilla afectados por pudrición basal. La caracterización molecular se realizó en diecisiete aislamientos asociados con la enfermedad, colectados en zonas productoras de pitahaya amarilla de los departamentos del Valle del Cauca, Risaralda, Boyacá y Cundinamarca, en Colombia. Se usó PCR con los cebadores ITS1 - ITS4 y TEF1α. Las secuencias de los productos de PCR se compararon con las registradas en el Cento Nacional de Información Biotecnológica (NCBI, por sus siglas en inglés), y se alinearon usando Clustal W2, se construyeron las relaciones filogenéticas con el programa MEGA 6 con el coeficiente de máxima verosimilitud. Los resultados de secuenciación para los marcadores mostraron que los aislamientos patogénicos correspondieron a F. oxysporum. El concepto filogenético confirmó que la pudrición basal de la pitahaya amarilla fue causada por F. oxysporum. Se identificó además una especie de Fusarium no reportada en el GenBank. AbstractThe aim of this study was to identify pathogenic isolates of Fusarium in fruits of yellow pitahaya affected by basal rot. Molecular characterization was performed on seventeen isolates associated with the disease, collected in yellow pitahaya producing areas of Valle del Cauca, Risaralda, Boyacá, and Cundinamarca in Colombia. PCR with primers ITS1 - ITS4 and TEF1α was used. The sequences of the PCR products were compared with the ones registered in National Center for Biotechnology Information (NCBI) and were aligned using Clustal W2 program, the phylogenetic relationships were built with MEGA6 program with the coefficient of maximum likelihood. The sequencing results for the markers showed that pathogenic isolates were F. oxysporum. Phylogenetic concept confirmed that basal rot of yellow pitahaya was caused by F. oxysporum. A species of Fusarium not reported in GenBank was also identified.

The evolutionary dynamics of transposable elements (TEs) are still poorly understood. One reason is that TE abundance needs to be studied at the population level, and despite recent advances in sequencing technologies, characterizing TE abundance in multiple populations by sequencing individuals separately is still too expensive. While sequencing pools of individuals (Pool-Seq) dramatically reduces sequencing costs, a comparison of TE abundance between pooled samples has been difficult, if not impossible, due to various biases. Here, we introduce a novel bioinformatic tool, PoPoolationTE2, which is specifically tailored for the comparison of TE abundance among pooled population samples or different tissues. Using computer simulations we demonstrate that PoPoolationTE2 not only faithfully recovers TE insertion frequencies and positions but, by homogenizing the power to identify TEs acrosss samples, it provides an unbiased comparison of TE abundance between pooled population samples. We anticipate that PoPoolationTE2 will greatly facilitate the analysis of TE insertion patterns in a broad range of applications.

Wild boar rooting is considered one of the main large soil disturbances affecting the structure and composition of plant communities in alpine grasslands. While direct consequences on plant community have been widely studied, their effects on soil seed banks have received little attention although rooting is assumed to determine the successional processes and ultimately the ecological recovery of the communities. The aim of this study is to assess the effect of wild boar rooting on species and community soil seed banks. The rooting effect was studied in terms of seed abundance and diversity in the most disturbed plant communities by wild boar in Pyrenean alpine grasslands. Two hundred soil core samples were collected at two depths to account for (short/long term) persistent and transient seed banks within and outside wild boar rooting in those grasslands. The ‘seedling emergence method’ was used to identify and quantify seeds from the germinable seed bank. The soil seed bank found within disturbances was smaller than expected. At the species level, the main type of seed bank represented in these grasslands was long-term persistent, comprising 75% of the seeds found. Rooting turned long-term persistent seed bank into short-term persistent and transient, by exposing seeds from the deepest part of the soil to germination conditions at the surface of disturbed areas. At the community level we found that rooting homogenized soil seed banks by increasing seed abundance and species richness in species-poor communities, while decreasing seed species dominance in nitrophilous communities. These results suggest a deep alteration of the structure of seed banks by wild boar rooting, which in turn, may not represent a real chance for colonization from soil seed banks.

ALTER is an open web-based tool to transform between different multiple sequence alignment formats. The originality of ALTER lies in the fact that it focuses on the specifications of mainstream alignment and analysis programs rather than on the conversion among more or less specific formats. In addition, ALTER is capable of identify and remove identical sequences during the transformation process. Besides its user-friendly environment, ALTER allows access to its functionalities in a programmatic way through a Representational State Transfer web service. ALTER’s front-end and its API are freely available at http://sing.ei.uvigo.es/ALTER/ and http://sing.ei.uvigo.es/ALTER/api/, respectively.

Summary The melanogenic marine bacterium Marinomonas mediterranea synthesizes a novel antimicrobial protein (LodA) with lysine-epsilon oxidase activity (EC 1.4.3.20). Homologues to LodA have been detected in several Gram-negative bacteria, where they are involved in biofilm development. Adjacent to lodA is located a second gene, lodB, of unknown function. This genomic organization is maintained in all the microorganisms containing homologues to these genes. In this work we show that lodA and lodB constitute an operon. Western blot analysis and enzymatic determinations revealed that LodA is secreted to the external medium when the culture reaches the stationary phase. LodB, on the other hand, has only been detected inside cells, but it is not secreted. The expression of the lysine-epsilon oxidase (LOD) activity in M. mediterranea requires functional copies of both genes since mutants lacking either lodA or lodB do not show any LOD activity. The active form of LodA containing the quinonic cofactor is intracellularly generated in a process that takes place only in the presence of LodB, suggesting that the latter is involved in this process. Moreover, in the absence of one of the proteins, the stability of the partner protein is compromised leading to a marked decrease in its cellular levels.

Aims We aimed to find out how a geophyte, Merendera montana, occupies a wide ecological, elevational and climatic range. This occurrence in that environmental array is outstanding not only among geophytes but also in the whole flora of the Iberian Peninsula, where only a very few plants show such an apparent success. Specifically, we compared morphological and reproductive traits, and frequency of different types of reproduction in disturbed and undisturbed grasslands. Furthermore, we have analyzed the phenology of this plant by trying to understand how it manages to spread in an exceptional elevational gradient of 2000 m, with contrasting climatic regimes between the mediterranean and the alpine regions. Methods We measured the plant density of M.montana populations in different types of Pyrenean grasslands either with or without small-scale disturbances at seven sites from the basal to the alpine belt (400– 2300 m a.s.l.). In each one of these populations, 100 individuals— 1200 in total—were uprooted to measure their morphological features as well as type and occurrence of reproduction. Phenology (flowering and fruiting dates and leaf lifespan) was estimated using 5 years of records in the studied areas and >200 herbarium sheets from the whole altitudinal rank and collected in the last 30 years. Differences in plant densities were analyzed with non-parametric Mann–Whitney U-test; differences in morphological traits, fruit and seed production associated with disturbance, with one-way analysis of covariance test (general linear model). Finally, linear regression analyses were used to determine the relationships between clonal reproduction and elevation and those between flowering, fruiting and senescence and date and elevation. Important Findings In all cases, plant density in disturbed grasslands was significantly higher than in undisturbed grasslands. Plant height and weight, bulb depth, leaf width and bulb and root weight were higher in disturbed plots. Disturbed and undisturbed plots were similar in most aspects of sexual reproduction, including fruiting percentage and seed production, but average seed weight was higher in the disturbed plots. Clonal reproduction and the synchrony of both types of reproduction were significantly higher in disturbed plots. Flowering and leaf emergence dates were not affected by disturbance but at the lowest elevations, they happened at least 2 months later than at the highest elevations. Summarizing, clonal reproduction, advantage in seedling establishment and an unusual but favorable phenology are the main factors in explaining the success of this geophyte colonizing disturbed soils that, in consequence, allow M.montana to settle in a wide range of climatic and ecological conditions.

DNA double strand breaks are the most cytotoxic lesions that can occur on the DNA. They can be repaired by different mechanisms and optimal survival requires a tight control between them. Here we uncover protein deneddylation as a major controller of repair pathway choice. Neddylation inhibition changes the normal repair profile toward an increase on homologous recombination. Indeed, RNF111/UBE2M-mediated neddylation acts as an inhibitor of BRCA1 and CtIP-mediated DNA end resection, a key process in repair pathway choice. By controlling the length of ssDNA produced during DNA resection, protein neddylation not only affects the choice between NHEJ and homologous recombination but also controls the balance between different recombination subpathways. Thus, protein neddylation status has a great impact in the way cells respond to DNA breaks. España , Ministerio de Economía y Competitividad SAF2010-14877

The melanogenic marine bacterium M. mediterranea synthesizes marinocine, a protein with antibacterial activity. We cloned the gene coding for this protein and named it lodA [P. Lucas–Elío, P. Hernández, A. Sanchez-Amat, F. Solano, Purification and partial characterization of marinocine, a new broad-spectrum antibacterial protein produced by Marinomonas mediterranea. Biochim. Biophys. Acta 1721 (2005) 193–203; P. Lucas-Elío, D. Gómez, F. Solano, A. Sanchez-Amat, The antimicrobial activity of marinocine, synthesized by M. mediterranea, is due to the hydrogen peroxide generated by its lysine oxidase activity. J. Bacteriol. 188 (2006) 2493–2501]. Now, we show that this protein is a new type of lysine oxidase which catalyzes the oxidative deamination of free l-lysine into 6-semialdehyde 2-aminoadipic acid, ammonia and hydrogen peroxide. This new enzyme is compared to other enzymes related to lysine transformation. Two different groups have been used for comparison. Enzymes in the first group lead to 2-aminoadipic acid as a final product. The second one would be enzymes catalyzing the oxidative deamination of lysine releasing H2O2, namely lysine-α-oxidase (LαO) and lysyl oxidase (Lox). Kinetic properties, substrate specificity and inhibition pattern show clear differences with all above mentioned lysine-related enzymes. Thus, we propose to rename this enzyme lysine-ε-oxidase (lod for the gene) instead of marinocine. Lod shows high stereospecificity for free l-lysine, it is inhibited by substrate analogues, such as cadaverine and 6-aminocaproic acid, and also by β-aminopropionitrile, suggesting the existence of a tyrosine-derived quinone cofactor at its active site. Abbreviations AANaminoacetonitrileβAPNβ-aminopropionitrileLAOl-amino acid oxidaseLαOLysine-α-oxidaseLatLysine-ε-aminotransferaseLodMarinocine (Lysine-ε-oxidase)LoxLysyl oxidaseLTQLysyl-Tyrosyl-QuinoneSDHsaccharopine dehydrogenaseSSAOsemicarbazide sensitive amino oxidaseTDQCtyrosine-derived quinone cofactorsTLCThin layer chromatography

Marinocine is a broad-spectrum antibacterial protein synthesized by the melanogenic marine bacterium Marinomonas mediterranea . This work describes the basis for the antibacterial activity of marinocine and the identification of the gene coding for this protein. The antibacterial activity is inhibited under anaerobic conditions and by the presence of catalase under aerobic conditions. Marinocine is active only in culture media containing l -lysine. In the presence of this amino acid, marinocine generates hydrogen peroxide, which causes cell death as confirmed by the increased sensitivity to marinocine of Escherichia coli strains mutated in catalase activity. The gene coding for this novel enzyme was cloned using degenerate PCR with primers designed based on conserved regions in the antimicrobial protein AlpP, synthesized by Pseudoalteromonas tunicata , and some hypothetical proteins. The gene coding for marinocine has been named lodA , standing for lysine oxidase, and it seems to form part of an operon with a second gene, lodB , that codes for a putative dehydrogenase flavoprotein. The identity of marinocine as LodA has been demonstrated by N-terminal sequencing of purified marinocine and generation of lodA mutants that lose their antimicrobial activity. This is the first report on a bacterial lysine oxidase activity and the first time that a gene encoding this activity has been cloned.

The relationships between a geophyte (Merendera montana (L.) Lange – Liliaceae) and a mole-vole (Microtus duodecimcostatus de Selys-Longchamps) in the Spanish Pyrenees were investigated by analysing plant density, asexual reproductive strategies, and chemical composition of M. montana, and by observing the feeding behavior of M. duodecimcostatus in captivity. We found that M. montana contains toxic alkaloids, the concentration of which varies throughout the year; being minimal at the end of the vegetative period. Alkaloids are stored in the whole plant, particularly in the leaves which are scarcely consumed by herbivores. Nevertheless, mole-voles eat this plant profusely both in the field and in captivity, where they showed preference for the corm. The corm has lower alkaloid concentration and higher levels of energetic substances than the other plant parts, particularly leaves. Although corm consumption causes plant death, M. montana grows more abundantly in areas colonized by mole-voles than in undisturbed grasslands with high plant cover and absence of mole-vole populations. Both asexual reproduction and seedlings of this species are more frequent in highly disturbed areas. Results strongly suggest a protocooperative relationship between mole-voles and this geophyte: the burrowing behavior of mole-voles favours dispersal and reproductive success of the plant, enhancing its habitat availability; the plant in turn supplies abundant and nutritious food at a low and "acceptable" toxicity cost.

Merendera montana (Liliaceae) is an endemic plant broadly, but exclusively, occurring in the Iberian Peninsula where it is one of the most abundant species in rangelands grazed by sheep and on the routes used by migrating sheep herds. In spite of showing a peculiar phenology adapted to Mediterranean regions (leaves remain green from fall to spring and are absent in summer), this species has also successfully colonized high altitude environments (up to 2300 m), and abundant populations can be found in montane and subalpine pastures. Chemical analysis has shown a high forage quality as well as the pre- sence of tropolone alkaloids (similar to other species of the genus Merendera), mainly colchicine, 3-demethylcolchicine, and colchicoside. In general terms, the alkaloid content was significantly lower in the subterranean corms than in the leaves. No remarkable seasonal variations in total alkaloids was found, although an increase of colchicoside in the corms and a decrease of colchicine was noted in summer. Both chemical deterrents and phenology could be the cause of the widespread ecological adaptation of Merendera montana. The alkaloid content would prevent intake by wild herbivores during winter, while the absence of aerial organs in summer, coinciding exactly with the highest domestic herbivore pressure, would permit this species to colonize rich soil habitats where there is low plant competition.

Three species of Microtus (voles) living in Pyrenean subalpine pastures brought seeds to the surface on vole mounds. This activity was studied to learn about the voles' influence on seed composition and vegetation gap colonization. Twentytwo plant species germinated from seed in the soil mounds. The plant composition of pastures was dominated by graminoids-hemicryptophytes. Nearly half of the seeds that occurred in mounds are of annual dicots able to quickly colonize the gaps and complete their vegetative cycle before the entry of perennial plants. Mound revegetation occurs chiefly by means of vegetative reproduction strategies. Nevertheless germination of seed brought to the surface can be important at the early stages of colonization after soil disturbance. Thus, such disturbances increase short-term plant diversity and patch heterogeneity.

Editorial.

The regulation of gene expression by microRNAs (miRNAs) is critical for normal development and physiology. Conversely, miRNA function is frequently impaired in cancer, and other pathologies, either by aberrant expression of individual miRNAs or dysregulation of miRNA synthesis. Here, we have investigated the impact of global disruption of miRNA biogenesis in primary fibroblasts of human or murine origin, through the knockdown of DGCR8, an essential mediator of the synthesis of canonical miRNAs. We find that the inactivation of DGCR8 in these cells results in a dramatic antiproliferative response, with the acquisition of a senescent phenotype. Senescence triggered by DGCR8 loss is accompanied by the upregulation of the cell-cycle inhibitor p21CIP1. We further show that a subset of senescence-associated miRNAs with the potential to target p21CIP1 is downregulated during DGCR8-mediated senescence. Interestingly, the antiproliferative response to miRNA biogenesis disruption is retained in human tumor cells, irrespective of p53 status. In summary, our results show that defective synthesis of canonical microRNAs results in cell-cycle arrest and cellular senescence in primary fibroblasts mediated by specific miRNAs, and thus identify global miRNA disruption as a novel senescence trigger. © 2013 The Anatomical Society and John Wiley & Sons Ltd., This work was supported by grants SAF2009-09031 and SAF2012-32117 from the Spanish Government to IP, and core support from MRCand grants from MRCT, CRUK, AICR, and the EMBO Young Investigator Programme to JG. We are grateful to Manuel Serrano and Daniel Peeperfor reagents and advice, and to Pablo Huertas for help with DDR assays.