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Native to the southeastern United States, the honey locust (Gleditsia triacanthos) is an invasive tree in several South American countries. Since eradication of invasive species is often costly, prevention is a better strategy. The relationship between invasive species and their habitats can be analyzed using species distribution models to produce maps of areas prone to the invasions. These maps can be used to develop efficient early detection plans of exotic species colonization. Here, we employed the Favorability Function model to assess the effects of environment and human activities on the invasive process of the honey locust in Uruguay. By integrating environmental and anthropic factors in our models, we obtained the best fitted prediction and classification indices. We showed that the southwestern region of the country concentrates the largest proportion of areas prone to the invasions. Environment was the main factor explaining the invasion of G. triacanthos, but the effect of human-related factors had a greatest effect in combination with environmental variables than on its own. We generated favorable risk maps and explanatory variables that can be used to more efficiently plan efforts to control the spread of this invasive.

Stearoyl-ACP desaturases (SADs) and fatty acid desaturases (FADs) play a critical role in plant lipid metabolism and also affect oil fatty acid composition introducing double bonds into the hydrocarbon chains to produce unsaturated fatty acids. In the present study, the genomic sequences of three SAD and three FAD candidate genes were characterized in olive and their expression was evaluated in different plant tissues. OeSAD genes corresponded to olive SAD1 and SAD2 and to a newly identified OeSAD4, sharing the conserved protein structure with other plant species. On the other hand, the full-length genomic sequences of two microsomal OeFAD genes (FAD2-1 and FAD2-2) and the plastidial FAD6, were released. When the level of expression was tested on different tissues of cv. Leccino, OeSAD1 and OeSAD2 were mainly expressed in the fruits, while OeFAD genes showed the lowest expression in this tissue. The mRNA profiling of all genes was directly studied in fruits of Leccino and Coratina cultivars during fruit development. In both genotypes, the expression level of OeSAD1 and OeSAD2 had the highest value during and after the pit-hardening period, when oil accumulation in fruit mesocarp is intensively increasing. Furthermore, the expression level of both OeFAD2 genes, which were the main candidates for oleic acid desaturation, were almost negligible during fruit ripening. These results have made possible to define candidate genes of the machinery regulation of fatty acid composition in olive oil, providing information on their sequence, gene structure and chromosomal location.

High fidelity during protein synthesis is accomplished by aminoacyl-tRNA synthetases (aaRSs). These enzymes ligate an amino acid to a cognate tRNA and have proofreading and editing capabilities that ensure high fidelity. Phenylalanyl-tRNA synthetase (PheRS) preferentially ligates a phenylalanine to a tRNAPhe over the chemically similar tyrosine, which differs from phenylalanine by a single hydroxyl group. In bacteria that undergo exposure to oxidative stress such as Salmonella enterica serovar Typhimurium, tyrosine isomer levels increase due to phenylalanine oxidation. Several residues are oxidized in PheRS and contribute to hyperactive editing, including against mischarged Tyr-tRNAPhe, despite these oxidized residues not being directly implicated in PheRS activity. Here, we solve a 3.6 Å cryo-electron microscopy structure of oxidized S. Typhimurium PheRS. We find that oxidation results in widespread structural rearrangements in the β-subunit editing domain and enlargement of its editing domain. Oxidization also enlarges the phenylalanyl-adenylate binding pocket but to a lesser extent. Together, these changes likely explain why oxidation leads to hyperaccurate editing and decreased misincorporation of tyrosine. Taken together, these results help increase our understanding of the survival of S. Typhimurium during human infection.

Mycotoxins have several toxicological implications. In the present study, we evaluate the presence of aflatoxin B1 (AFB1), ochratoxin A (OTA), and fumonisin (FB1) in paddy rice, polished rice, and maize from the fields and markets in Nayarit State (Mexico). The results indicated the presence of AFB1 in 21.21% of paddy rice samples and 11.11% of market maize samples. OTA was present in only 3.03% (one sample) of paddy rice samples. FB1 was detected in 87.50% and 88.88% of maize samples from field and market, respectively. The estimated human exposure was calculated for FB1 using the probable daily intake (PDI), which suggested that FB1 could contribute to the development of diseases through the consumption of contaminated maize. Positive samples indicated that some rice and maize samples were not suitable for human consumption. Further efforts are needed to continue monitoring mycotoxins and update national legislation on mycotoxins accordingly This work was supported by the Consejo Nacional de Ciencia y Tecnolgía (CONACYT) under Grant code SALUD-2017-2-289726.

The loss of individuals in disturbed sites may be buffered by the presence of small natural features (SNFs), sites acting as ecological refuges preventing the loss of phenotypic variability and genetic diversity of plants. An important group of SNFs is rocky outcrops, singular geological formations that stand out from the surrounding matrix which could host a greater species diversity. We tested the value of rocky outcrops as SNFs of a Tertiary relict tree, the Chilean lucuma Pouteria splendens (Sapotaceae). We identified vegetation patches with P. splendens and characterized their landscape properties in sites associated with rocky outcrops and clearings (plains). Then, we contrasted the genetic diversity, phenotypic variability and sapling abundance of P. splendens inhabiting rocky outcrops and plains. We observed that rocky outcrops sustained subtle higher genetic diversity compared with plains, but non-significant genetic structuration was detected between habitat types. Independently of the habitat type, P. splendens have a significant role in the sapling recruitment: their number was higher underneath of conspecifics than in open spaces. Besides, plants in the rocky outcrops produced smaller and ovoid-shaped fruits, as a possible result of selective pressures generated by frugivorous. We propose that rocky outcrops are regeneration sites for P. splendens. However, subtle differences in genetic diversity between habitats and the lacking of genetic structuration suggest that conservation strategies should be focused on the protection of all habitat types as a manner of maintaining ecosystem processes in this singular vegetation remnant.

Eriosyce is one of most species‐rich genera within Notocacteae (Cactaceae) harboring a variety of stem and flower morphologies, and fruits with basal abscission. The lack of a well‐sampled molecular phylogeny contributes to the current taxonomic instability of the genus, where its circumscription and infrageneric classification has been questioned. Specimens of Eriosyce (63 taxa) plus 19 outgroups were analyzed through sequencing three plastid noncoding introns (rpl32‐trnL, trnL‐trnF, trnH‐psbA), one plastid gene (ycf1), and one nuclear gene (PHYC). Individual markers and concatenated matrices were analyzed using maximum likelihood and Bayesian approaches. Phylogenetic analyses strongly support the monophyly of Eriosyce s.l. Furthermore, seven clades within Eriosyce s.l. were defined based on supported branches, although one of them was weakly supported. Our results suggest that some past taxonomic proposals have low phylogenetic support and should no longer be used, e.g., based on their scattered positions in the phylogenetic reconstruction, several infraspecific taxa appear unrelated to the typical form of the species in which they had been placed. We present a phylogeny‐informed infrageneric classification of the genus Eriosyce, and new combinations are proposed to update the nomenclature of species and sections.

Delayed flowering due to later snowmelt and colder temperatures at higher elevations in the alpine are expected to lead to flowering phenological adjustment to prevent decoupling of peak flowering from the warmest time of the year, thereby favoring pollination. However, even if flowering is brought forward in the season at higher elevations, an elevational temperature gap is likely to remain between the high- and low-elevation populations of a species at the time these reach peak flowering on account of the atmospheric reduction in temperature with increasing elevation. The negative effect of this temperature gap on pollination could be compensated by plastically-prolonged flower life spans at higher elevations, increasing the probability of pollination. In a tightly temperature-controlled study, the flowering phenology adjustment and flower longevity compensation hypotheses were investigated in an alpine species in the Andes of central Chile. The snow free period varied from 7 to 8.2 months over 810 m elevation. Temperatures were suitable for growth on 82–98% of the snow free days. Flowering onset was temporally displaced at the rate of 4.6 d per 100 m increase in elevation and flowering was more synchronous at higher elevations. Flowering phenology was adjusted over elevation. The latter was manifest in thermal sums tending to decrease with elevation for population flowering onset, 50% flowering, and peak flowering when the lower thermal limit for growth (TBASE) was held constant over elevation. For TBASE graded over elevation so as to reflect the growing season temperature decline, thermal sums did not vary with elevation, opening the door to a possible elevational decline in the thermal temperature threshold for growth. Potential flower longevity was reduced by passive warming and was more prolonged in natural populations when temperatures were lower, indicating a plastic trait. Pollination rates, as evaluated with the Relative Pollination Rate index (RPR), when weighted for differences in floral abundance over the flowering season, declined with elevation as did fruit set. Contrary to expectation, the life-spans of flowers at higher elevations were not more prolonged and failed to compensate for the elevational decrease in pollination rates. Although strong evidence for phenological adjustment was forthcoming, flower longevity compensation did not occur over Oxalis squamata´s elevational range. Thus, flower longevity compensation is not applicable in all alpine species. Comparison with work conducted several decades ago on the same species in the same area provides valuable clues regarding the effects of climate change on flowering phenology and fitness in the central Chilean alpine where temperatures have been increasing and winter snow accumulation has been declining.

Caulobacter crescentus is a Gram-negative alphaproteobacterium that commonly lives in oligotrophic fresh- and saltwater environments. C. crescentus is a host to many bacteriophages, including ϕCbK and ϕCbK-like bacteriophages, which require interaction with the bacterial flagellum and pilus complexes during adsorption. It is commonly thought that the six paralogs of the flagellin gene present in C. crescentus are important for bacteriophage evasion. Here, we show that deletion of specific flagellins in C. crescentus can indeed attenuate ϕCbK adsorption efficiency, although no single deletion completely ablates ϕCbK adsorption. Thus, the bacteriophage ϕCbK likely recognizes a common motif among the six known flagellins in C. crescentus with various degrees of efficiency. Interestingly, we observe that most deletion strains still generate flagellar filaments, with the exception of a strain that contains only the most divergent flagellin, FljJ, or a strain that contains only FljN and FljO. To visualize the surface residues that are likely recognized by ϕCbK, we determined two high-resolution structures of the FljK filament, with and without an amino acid substitution that induces straightening of the filament. We observe posttranslational modifications on conserved surface threonine residues of FljK that are likely O-linked glycans. The possibility of interplay between these modifications and ϕCbK adsorption is discussed. We also determined the structure of a filament composed of a heterogeneous mixture of FljK and FljL, the final resolution of which was limited to approximately 4.6 A. Altogether, this work builds a platform for future investigations of how phage ϕCbK infects C. crescentus at the molecular level. IMPORTANCE Bacterial flagellar filaments serve as an initial attachment point for many bacteriophages to bacteria. Some bacteria harbor numerous flagellin genes and are therefore able to generate flagellar filaments with complex compositions, which is thought to be important for evasion from bacteriophages. This study characterizes the importance of the six flagellin genes in C. crescentus for infection by bacteriophage ϕCbK. We find that filaments containing the FljK flagellin are the preferred substrate for bacteriophage ϕCbK. We also present a high-resolution structure of a flagellar filament containing only the FljK flagellin, which provides a platform for future studies on determining how bacteriophage ϕCbK attaches to flagellar filaments at the molecular level.

This data paper presents a largely phylogeny-based online taxonomic backbone for the Cactaceae compiled from literature and online sources using the tools of the EDIT Platform for Cybertaxonomy. The data will form a contribution of the Caryophyllales Network for the World Flora Online and serve as the base for further integration of research results from the systematic research community. The final aim is to treat all effectively published scientific names in the family. The checklist includes 150 accepted genera, 1851 accepted species, 91 hybrids, 746 infraspecific taxa (458 heterotypic, 288 with autonyms), 17,932 synonyms of accepted taxa, 16 definitely excluded names, 389 names of uncertain application, 672 unresolved names and 454 names belonging to (probably artificial) named hybrids, totalling 22,275 names. The process of compiling this database is described and further editorial rules for the compilation of the taxonomic backbone for the Caryophyllales Network are proposed. A checklist depicting the current state of the taxonomic backbone is provided as supplemental material. All results are also available online on the website of the Caryophyllales Network and will be constantly updated and expanded in the future. Citation: Korotkova N., Aquino D., Arias S., Eggli U., Franck A., Gomez-Hinostrosa C., Guerrero P. C., Hernandez H. M., Kohlbecker A., Kohler M., Luther K., Majure L. C., Muller A., Metzing D., Nyffeler R., Sanchez D., Schlumpberger B. & Berendsohn W. G. 2021: Cactaceae at Caryophyllales.org – a dynamic online species-level taxonomic backbone for the family. – Willdenowia 51: 251–270. Version of record first published online on 31 August 2021 ahead of inclusion in August 2021 issue. Data published through: http://caryophyllales.org/cactaceae/Checklist

The increasing demand of natural products for the control of plant diseases has led to the search for biological control agents, namely fungi, often isolated from composts of organic residues. By composting two different mixes of agro-industrial residues, P1 and P2 composts were obtained, from where a known antagonist, Trichoderma atroviride, was isolated. Later, a second composting process was performed and when the composting mix reached room temperature, the previously isolated T. atroviride was inoculated in both composts to enrich its population (E) and named as P1E and P2E. The suppressive capacity of these two composts against Sclerotium rolfsii, Clarireedia spp. and Rhizoctonia solani was tested two weeks after T. atroviride inoculation, in 100 mL pots with turfgrass seeded with Agrostis stolonifera. The tested treatments were: composts P1 and P2 without any treatment; thermally treated P1 and P2 (P1t, P2t); P1 and P2 enriched without and with previous thermic treatment (P1E, P2E, P1tE, P2tE), and a commercial peat-based substrate, natural (P) and thermally treated (Pt), enriched (PE) and enriched after thermal treatment (PtE). Enriched composts achieved the highest disease control. P1E was more effective in controlling Clarireedia spp.; pots with infected plants grown on P1E, showed 53.5% less affected area compared to P1t. P2E was more effective against R. solani; results showed 69.3% less affected area compared to P2t and both were effective in S. rolfsii control, with a reduction of 38.5% in P1E and 43.5% in P2E. The larger population of Trichoderma spp. observed in the enriched substrates associated to the greater enzyme activity, namely and N-acetyl-β-glucosaminidase, standing out in P2E, will have contributed strongly to the biocontrol of the studied diseases. An increase in some macro and micronutrients was observed in the enriched heat-treated substrates. FCT: SFRH/BD/109218/2015 info:eu-repo/semantics/publishedVersion

The use of pig deep-litter (PDL) in pasture fertilization can be an important alternative from an environmental and economic point of view. This study was conducted to evaluate the fertilization with PDL exclusively or in association with mineral fertilizers on the quality and productivity of Panicum maximum cv. Mombasa. The experiment was carried out in Ipameri, GO, Brazil. The design was a randomized block design in a 2 x 5 factorial scheme with four repetitions, in plots of 24 m². The doses of PDL (0; 5; 10; 15 and 20 Mg ha-1) were tested, with or without fertilization with NPK (50 kg ha-1 of P2O5, 10 kg ha-1 of K2O and 10 kg ha-1 of N). The organic fertilizer was applied broadcast at the planting of the pasture, and the mineral fertilizers were split and applied at planting and as topdressing. Dry mass, crude protein, acid detergent fiber and neutral detergent fiber were evaluated. PDL doses associated with mineral fertilization led to linear increases from 52 to 282% in the analyzed variables and better bromatological composition. PDL application is beneficial to the production of pastures, and increasing applications up to the PDL dose of 20 Mg ha-1 associated with mineral fertilization promoted better quantitative and qualitative results. With exclusive fertilization with PDL, the gains in quality and productivity were more modest. RESUMO O uso de cama sobreposta de suínos (CSS) na adubação de pastagens pode ser uma importante alternativa do ponto de vista ambiental e econômico. Com o objetivo de avaliar a adubação com CSS de forma exclusiva ou associada a fertilizantes minerais, na qualidade e produtividade de Panicum maximum cv. Mombaça, foi conduzido um experimento em condições de campo, em Ipameri, GO. O delineamento foi em blocos ao acaso em esquema fatorial 2 x 5 com quatro repetições, em parcelas de 24 m². Foram testadas as doses de CSS (0; 5; 10; 15 e 20 Mg ha-1), com ou sem adubação com NPK (50 kg ha-1 de P2O5, 10 kg ha-1 de K2O e 10 kg ha-1 de N). O adubo orgânico foi aplicado a lanço na implantação da pastagem, e os fertilizantes minerais parcelados no plantio e em cobertura. Foram avaliadas a massa seca, proteína bruta, fibra em detergente ácido e fibra em detergente neutro. As doses de CSS associadas à adubação mineral apresentaram incrementos lineares de 52 a 282% nas variáveis avaliadas, e melhor composição bromatológica. A aplicação de CSS é benéfica à produção de pastagens e as aplicações crescentes até à dose de 20 Mg ha-1 de CSS associadas à adubação mineral apresentaram melhores resultados quantitativos e qualitativos. Com a adubação exclusiva com CSS os ganhos de qualidade e produtividade do capim Mombaça são menores.

Algarve region (Portugal) has nearly 40 golf courses with a significant economic impact. Summer surveys on 10 golf courses detected an unknown disease on one course in 2009 and on another course in 2012 and 2013 at 29 to 30°C daily average. The second course had symptoms on about 25% of the turf of two putting greens. Diseased bentgrass (Agrostis stolonifera L.) had a green dappled pattern with irregular patches of turfgrass on yellowed leaves. Prior to decaying, affected leaves turned brown and then gray. Crown and leaf sheath infections resulted in dark brown dry rot. No lesions were observed on the roots. Leaves were surface disinfected with 5% commercial bleach (0.225% sodium hypochlorite) and cultured on potato dextrose agar (PDA). Ten fungal colonies grew from the leaf tissue, and brown mycelia, conidiophores, and conidia were observed under a microscope. Conidia were ventricose pyriform, mostly abruptly curved, 20 to 36 µm (30 µm, SD = 4) × 7 to 12 µm (10.5 µm, SD = 1.3) (n = 50), predominantly three-septate, with a prominent hilum and enlarged and darkened central cells. Colonies grown on PDA were black-brown with a black reverse side. Conidia differed in size, 15.4 to 24.6 µm (19.99 µm, SD = 3.00) × 6 to 11 μm (8.68 µm, SD = 1.54) (n = 50) and morphology (cylindrical or slightly curved). These characteristics were consistent with Curvularia trifolii (Kauffm.) Boedijn. (Ellis 1971; Falloon 1976; Khadka 2016). Species identification of the representative isolate A2 1.12 was confirmed by analysis of nucleotide sequences of the ITS1-5.8S-ITS2 region using primers ITS1 and ITS4 (White et al. 1990) and GPDH gene region with primer set gpd (Koike et al. 2013). BLAST searches of GenBank showed a high similarity of the isolate ITS sequence (MG029439) to the reference sequence JN712458 of C. trifolii (99% identity) and GPDH sequence (MK570108) with LT715803.1 (97.88% identity). The maximum likelihood phylogenetic tree showed that our isolate clustered with C. trifolii. The pathogenicity assay of this isolate was conducted in greenhouse on A. stolonifera ‘Penncross’. The isolate was grown on PDA (25°C, 10 days). Five pots (100 ml) were filled with a sand and peat mix (9:1 v/v) with 0.06 g of seeds per pot, covered with a fine sand layer. Turfgrass was cut once a week beginning 2 weeks after seeding and was fertigated with 0.5 g/liter of Peter’s foliar feed (27 + 15 + 12; N + P2O5 + K2O; and micronutrients; Scotts, Heerlen, The Netherlands). To obtain a conidial suspension for inoculation, cultured plates were scraped with a sterilized spreader and water. The suspension was filtered through a sterile gauze. Conidia were counted under a microscope (400×) with a hemocytometer. The suspension was adjusted to 8 × 103 conidia/ml, and 10 ml was sprayed per pot. Pots maintained humidity for 2 days under microtunnels. The first disease symptoms appeared 3 days after inoculation. Bentgrass from the five pots developed Curvularia blight and rotted crown symptoms. Control plants (five pots treated with water) did not display symptoms. This trial was repeated once. On PDA, C. trifolii was reisolated from leaf lesions and morphologically identified, confirming Koch’s postulates. Ellis (1971) referred to the presence of C. trifolii in Portugal, but no region, symptom description, or grass species was detailed. Sivanesan (1987) reported C. trifolii in Portugal only on Lolium multiflorum. Therefore, this is the first report of C. trifolii in Algarve, affecting A. stolonifera. This disease can increase maintenance costs in greens in this area. info:eu-repo/semantics/publishedVersion

How does microbiota research impact our understanding of biological individuality? We summarize the interdisciplinary summer school on "Microbiota, symbiosis and individuality: conceptual and philosophical issues" (July 2019), which was supported by a European Research Council starting grant project "Immunity, DEvelopment, and the Microbiota" (IDEM). The summer school centered around interdisciplinary group work on four facets of microbiota research: holobionts, individuality, causation, and human health. The conceptual discussion of cutting-edge empirical research provided new insights into microbiota and highlights the value of incorporating into meetings experts from other disciplines, such as philosophy and history of science. Video Abstract., Microbiome, 8 (1), ISSN:2049-2618

Populations of the columnar cactus genus Eulychnia (Cactaceae) are an iconic sight in the Chilean Atacama Desert. The most recent taxonomic treatment of the genus suggested to accept up to seven ta...

Over a period of eight years, were assessed the main types of airborne pollen in Mexico City (a tropical region). We established the airborne pollen calendar and we evaluated the associations between aeroallergens and allergic diseases in children treated at hospitals. A Hirst-type volumetric spore trap was placed in an open area within the Chapultepec Forest, and pollen records were obtained during the flowering period of the plants. A total of 502 patients between 1 and 17 years old were evaluated. The patients were diagnosed by skin testing with extracts of tree pollen, grass and weeds. During the monitoring period, the airborne pollen grains were dominated by trees (Fraxinus, Cupressaceae, Alnus, Quercus) followed by Poaceae, with the highest average values detected between January and February. Children with positive reactions to pollen aeroallergens were diagnosed with allergic diseases. The predominant airborne allergens were determined. An airborne pollen calendar was established. A close association was found between the most frequent and abundant pollen types and the responses of children produced by exposure to pollen grains. Children with positive reactions to pollen were diagnosed with allergic conjunctivitis, allergic rhinitis, asthma and keratoconjunctivitis.

Background Sympatric congeneric plants might share pollinators, or each species might avoid competition by evolving specialized traits that generate partitions in pollinator assemblages. In both cases, pollen limitation (a decrease in the quality and quantity of compatible reproductive pollen) can occur, driving the plant mating system to autogamy as a mechanism of reproductive assurance. We assessed the relationships between pollinator assemblages and mating systems in a group of sympatric congeneric plants. We attempted to answer the following questions: (i) How similar are pollinator assemblages among sympatric cactus species? (ii) Which mating systems do sympatric cactus species use? Methods We studied sympatric Eriosyce taxa that inhabit a threatened coastal strip in a mediterranean-type ecosystem in central Chile. We performed field observations on four taxa and characterized pollinators during the years 2016 and 2017. We estimated differences in the pollinator assemblages using the Bray–Curtis index. To elucidate the mating systems, we conducted hand-pollination experiments using three treatments: manual cross-pollination, automatic self-pollination, and control (unmanipulated individuals). We tested differences in seed production for statistical significance using Kruskal–Wallis analysis. Results Eriosyce subgibbosa showed a distinctive pollinator assemblage among the sympatric species that we studied (similarity ranged from 0% to 8%); it was visited by small bees and was the only species that was visited by the giant hummingbird Patagona gigas. Pollinator assemblages were similar between E. chilensis (year 2016 = 4 species; 2017 = 8) and E. chilensis var. albidiflora (2016 = 7; 2017 = 4); however, those of E. curvispina var. mutabilis (2016 = 7; 2017 = 6) were less similar to those of the aforementioned species. E. curvispina var. mutabilis showed the highest interannual variation in its pollinator assemblage (18% similarity). Reproduction in E. subgibbosa largely depends on pollinators, although it showed some degree of autogamy. Autonomous pollination was unfeasible in E. chilensis, which depended on flower visitors for its reproductive success. Both E. chilensis var. albidiflora and E. curvispina var. mutabilis showed some degree of autogamy. Discussion We observed differences in pollinator assemblages between E. subgibbosa and the remaining Eriosyce taxa, which depend on hymenopterans for pollen transfer. Pollinator assemblages showed considerable interannual variation, especially those of E. subgibbosa (ornithophilous syndrome) and E. curvispina var. mutabilis (melitophilous syndrome). Autogamous reproduction in these taxa may act as a reproductive assurance mechanism when pollinator availability is unpredictable. Our study contributes to improving our understanding of the reproductive systems of ecological interactions between threatened species in a Chilean mediterranean-type ecosystem.

This report describes the genome sequences of four Salmonella enterica subsp. enterica serovar Gallinarum ( Salmonella Gallinarum) strains isolated in Colombia in 2017 from layer breeders of different ages. The layer breeder flocks were presenting with an elevated mortality with lesions typical of fowl typhoid (FT).

Bright yellow, interveinal chlorosis was observed for the first time on leaves of the older and mid-growth of cucurbit plants in southern Arizona and Sonora (Mexico) during September and October of 2006. Some cultivars exhibited substantial yield losses of 30 to 80%. In Arizona, symptoms were in Cucumis melo (muskmelon and honeydew melon) fields in the Yuma Valley and Hyder. In Sonora, honeydew and muskmelon, Cucurbita pepo (acorn, spaghetti, and summer [yellow and zucchini] squash), and Citrullus lanatus (watermelon) were symptomatic in Hermosillo, whereas, in Caborca, honeydew and cantaloupe developed similar symptoms. Interveinal chlorosis was observed in 60 to 100% of the plants in each field. Crops planted mid-to-late season were 100% infected, whereas, the early-season fields experienced approximately 60 to 80% incidence. All symptomatic fields in the Sonoran Desert and vicinity were infested by the whitefly Bemisia tabaci (Genn.), which was identified as the ‘B biotype’ on the basis of mitochondria COI sequence analysis (data not shown). Whitefly population levels were variable and ranged from 5 to 200 per plant. Total RNA was isolated from leaf samples collected from symptomatic plants using Tri Reagent (Molecular Research Center, Cincinnati, OH). Purified RNA was used in reverse transcriptase-PCR with primers specific to the Cucurbit yellow stunting disorder virus (CYSDV) coat protein (CP) gene (RNA2-deoxyribonucleotide coordinates 4927-4950 and 5657-5679) for the suspected whitefly-transmitted bipartite CYSDV (4). PCR yielded the CYSDV CP fragment, at 753 bp (GenBank Accession Nos. EF21058 and EF21059), which was cloned into pGEM T-Easy and sequenced in both directions using universal primers. The CYSDV CP nucleotide sequences (n = 16) obtained from acorn squash, honeydew melon, muskmelon, yellow squash, and watermelon had 99 to 100% identity. The Arizona (AZ) and Sonora (SON) CYSDV CP sequences shared 99 to 100% identity with previously described CYSDV isolates from the Eastern Hemisphere (GenBank Accession Nos. DQ903105 and DQ903108) and also with two isolates of CYSDV collected during 2004 from Zacapa Valley, Guatemala (GenBank Accession Nos. EF21060 and EF21061) (J. K. Brown, unpublished data). CYSDV is a member of the genus Crinivirus, family Closteroviridae. CYSDV was first identified in cucumber and melon crops in the Middle East approximately 15 years ago and 10 years ago in Spain (1). Most recently, this virus was introduced into Texas (2), Guatemala (J. K. Brown, unpublished data), and Arizona and California (3). CYSDV has therefore emerged as an important and potentially worldwide threat to the production of cultivated cucurbits (3). The threat appears to be significant in light of the introduction or establishment of the exotic B. tabaci biotypes B and Q vectors, which also originated in the Middle Eastern-North African-Mediterranean region. To our knowledge, this is the first report of CYSDV infecting field-grown C. pepo (four types) and watermelon, reported previously only as experimental laboratory hosts, and of CYSDV in two types of melon (C. melo) in Mexico. References: (1) A. Celix et al. Phytopathology 86:1370, 1996. (2) J. Kao et al. Plant Dis. 84:101, 2000. (3) Y.-W. Kuo et al. Plant Dis. 91:330, 2007. (4) L. Rubio et al. J. Gen. Virol. 82:929, 2001.

Severe yellow leaf curl and plant stunting symptoms were observed in tomato plants from two home gardens in central Arizona (Phoenix area) and a tomato field in Sonora, Mexico during the fall of 2006. Disease symptoms were reminiscent of those reported in Florida during 1994 (4) and more recently in tomato fields in the Pacific Coast state of Sinaloa, Mexico found to be infected with the exotic Tomato yellow leaf curl virus (TYLCV) (2). Total DNA was extracted from two symptomatic tomato plants from Arizona and Sonora and used as a template in PCR. PCR products of the core region of the begomovirus coat protein gene (Cp) were cloned (n = 3) and the DNA sequence was determined. BLAST analysis of the 579 bases with sequences available in the NCBI GenBank database indicated the closest match was to an isolate of the monopartite begomovirus TYLCV from Israel, which was known to have been introduced into the Caribbean region, including Puerto Rico, the southeastern United States, and Mexico from 1990 to 1996 (1,4). The full-length TYLCV genome (approximately 2,800 bases) was amplified for a field isolate from each location by rolling circle amplification (RCA) using TempliPhi (Amersham Biosciences, Piscataway, NJ). RCA products were cloned into the plasmid vector pGEM7 (Promega, Madison, WI) that had been previously digested with SacI endonuclease. The complete TYLCV genome sequence was determined for six clones from each RCA product. Nucleotide analysis indicated that the complete TYLCV genome sequences from Sonora and Arizona, respectively, shared 97.6 and 97.7% nt identity. The comparative sequence analysis indicated that TYLCV-Sonora (TYLCV-Son) (GenBank Accession No. EF210555) was 99.1% nt identical to TYLCV reported recently from Culiacan, Mexico (GenBank Accession No. DQ631892). In contrast, TYLCV-AZ (GenBank Accession No. EF210554) shared 99.3% identity with an isolate from Texas, TYLCV-TX (GenBank Accession No. EF110890) (3). Interestingly, the TX and AZ TYLCV isolates contained a unique 29-nt deletion in the intergenic region (IR) between the TATA-box and the nonanucleotide, initiating at nt coordinate 2696. Except for the deletion in the IR region of the AZ and TX isolates, these viruses shared 97.6 to 99.1% nt identity to other TYLCV isolates reported in the Western Hemisphere. The genome sequence for TYLCV-Son shares high nt identity with TYLCV isolates identified in the Yucatan Peninsula and Pacific Coast of Mexico (2), the Caribbean region, and the southeastern United States, suggesting that a single TYLCV species was introduced and has spread throughout North America and the Caribbean (4). The absence of other TYLCV isolates in the Western Hemisphere with the novel 29-nt deletion noted for the TX and AZ isolates suggests that the latter two isolates originated from the same U.S. source. In Mexico, TYLCV was first introduced in the east coast and Yucatan region approximately in 1996. From there, this isolate has spread to the western part of the country (Sinaloa and Sonora) from 2004 to 2006 (2). Similarly, in the United States, TYLCV was introduced and spread in the eastern U.S. states beginning in 1994 (4), where it had been confined until it was discovered in Texas (3) and now Arizona during 2006. References: (1) J. Bird et al. Plant Dis. 85:1028, 2001. (2) J. K. Brown and A. M. Idris. Plant Dis. 90:1360, 2006. (3) T. Isakeit et al. Plant Dis. 91:466, 2007. (4) J. E. Polston et al. Plant Dis. 78:831, 1994.

Perennial cankers and consequent grapevine dieback are a major problem in vineyards of Sonora and Baja California, the most important grape-production areas of Mexico. In order to identify the canker-causing agents, symptomatic arms, cordons, and trunks were collected from 13 and 6 vineyards in Sonora and Baja California, respectively. Two Botryosphaeriaceae spp., Lasiodiplodia theobromae and Diplodia seriata, were isolated frequently from infected wood and identified based on morphological and cultural characters as well as analyses of nucleotide sequences of three genes, the internal transcribed spacer region (ITS1-5.8S-ITS2), a partial sequence of the β-tubulin gene, and part of the translation elongation factor 1-α gene (EF1-α). Although both L. theobromae and D. seriata were isolated from grapevine cankers in Baja California, only L. theobromae was found in vines in the Sonora region. Pathogenicity of both species was verified by inoculation of rooted cuttings and green shoots of Thompson Seedless and Chardonnay cultivars. Isolates of L. theobromae were more virulent, based on the extent of spread in the secondary wood and green tissue, than those of D. seriata. These findings confirm L. theobromae and D. seriata as the causal agents of dieback and canker formation of grapevines in northern Mexico.

Contractile injection systems (bacteriophage tails, type VI secretions system, R-type pyocins, etc.) utilize a rigid tube/contractile sheath assembly for breaching the envelope of bacterial and eukaryotic cells. Among contractile injection systems, bacteriophages that infect Gram-positive bacteria represent the least understood members. Here, we describe the structure of Listeria bacteriophage A511 tail in its pre- and post-host attachment states (extended and contracted, respectively) using cryo-electron microscopy, cryo-electron tomography, and X-ray crystallography. We show that the structure of the tube-baseplate complex of A511 is similar to that of phage T4, but the A511 baseplate is decorated with different receptor-binding proteins, which undergo a large structural transformation upon host attachment and switch the symmetry of the baseplate-tail fiber assembly from threefold to sixfold. For the first time under native conditions, we show that contraction of the phage tail sheath assembly starts at the baseplate and propagates through the sheath in a domino-like motion.

Common water disinfectants like chlorine have been reported to select for resistant viruses, yet little attention has been devoted to characterizing disinfection resistance. Here, we investigated the resistance of MS2 coliphage to inactivation by chlorine dioxide (ClO2). ClO2 inactivates MS2 by degrading its structural proteins, thereby disrupting the ability of MS2 to attach to and infect its host. ClO2-resistant virus populations emerged not only after repeated cycles of ClO2 disinfection followed by regrowth but also after dilution-regrowth cycles in the absence of ClO2. The resistant populations exhibited several fixed mutations which caused the substitution of ClO2-labile by ClO2-stable amino acids. On a phenotypic level, these mutations resulted in a more stable host binding during inactivation compared to the wild-type, thus resulting in a greater ability to maintain infectivity. This conclusion was supported by cryo-electron microscopy reconstruction of the virus particle, which demonstrated that ...

SUMMARYThe incidence of Mediterranean spotted fever (MSF) in Catalonia (Spain) has decreased in the last two decades. The prevalence of antibodies to Rickettsia conorii in human beings and dogs in the region of Vallès Occidental (Catalonia) was assessed by indirect immunofluorescence, and the results compared with those obtained in a similar study from 1987. Nineteen (5·0%) out of 383 human serum samples had antibodies to R. conorii. This seroprevalence was significantly lower (11·5%) (P = 0·003) than that recorded in the 1987 survey. Forty-two out (42·0%) of 100 canine serum samples had antibodies to R. conorii. A high proportion of the studied dogs (91·0%) were receiving anti-tick treatment, mainly with permethrin-imidacloprid spot-on (Advantix, Bayer, Germany). The current canine seroprevalence was not significantly different from that recorded in the 1987 survey (36.9%). In conclusion, this study shows a significant decrease in the prevalence of antibodies to R. conorii in the human population of Catalonia in the last 20 years, which corresponds with a decrease in the number of cases of MSF. We suggest that the widespread use of anti-tick treatment in dogs could limit the introduction of ticks to humans due to a reduction of infestation duration in dogs, thus contributing to the decrease in MSF incidence.

Several systems, including contractile tail bacteriophages, the type VI secretion system and R-type pyocins, use a multiprotein tubular apparatus to attach to and penetrate host cell membranes. This macromolecular machine resembles a stretched, coiled spring (or sheath) wound around a rigid tube with a spike-shaped protein at its tip. A baseplate structure, which is arguably the most complex part of this assembly, relays the contraction signal to the sheath. Here we present the atomic structure of the approximately 6-megadalton bacteriophage T4 baseplate in its pre- and post-host attachment states and explain the events that lead to sheath contraction in atomic detail. We establish the identity and function of a minimal set of components that is conserved in all contractile injection systems and show that the triggering mechanism is universally conserved.

Turfgrasses are affected by fungal diseases and their chemical control is currently limited by the legislation and the impact on the environment of fungicides and the possible development of fungicide resistance. This study was carried out during three years to evaluate the antagonistic effect of the microbiota associated with organic composts. Two composts were produced from agro-industrial residues, P1 and P2. Fungi from both composts were isolated and identified and their antagonistic capacity was evaluated, in vivo and in vitro tests, against Sclerotium rolfsii Sacc, Clarireedia gen. nov. (formerly Sclerotinia homoeocarpa (Benn, 1937)) and Rhizoctonia solani (Kuhn 1858). In vivo tests were run for Agrostis stolonifera L., seeded in P1, P2 and a commercial peat (T) based substrate (Hansa Torf Floragard, Germany), in 100 mL pots. A part of each substrate was submitted to a thermic treatment, obtained Pt, P1t and P2t substrates. Antagonistic properties of the P, P1 and P2 were compared with substrates with thermic treatment. In the in vitro tests Trichoderma atroviride and Trichoderma asperellum showed higher growth than the tested pathogenic agents. T. atroviride showed the best antagonistic capacity, with inhibition percentages of 67.5%, 63.8% and 62.5% against Clarireedia spp., S. rolfsii and R. solani, respectively. In the in vivo tests, the efficacy and biological control index (BCI) were higher in P1 and P2 than in P. BCI ranged from 1.66 (N) to 39.03 (P2) on S. rolfsii control, from 1.38 (Pt) to 14.66 (P1) on Clarireedia spp. control, and from 1.87 (Pt) to 21.45 (P1) for R. solani control. Compost P2 showed the highest suppressive effect on the studied soil diseases in turfgrass, which did not occur when its microbiota was eliminated by thermic treatment. Portuguese Foundation for Science and TechnologyPortuguese Foundation for Science and Technology [SFRH/BD/109218/2015] info:eu-repo/semantics/publishedVersion

Copiapoa (Cactaceae) is a genus endemic to the Chilean Atacama Desert. The taxa of Copiapoa subsection Cinerei occur in an area of high species richness and high levels of species endemism of the Central Chilean biodiversity hotspot. Four taxa are usually recognised in this group: Copiapoa gigantea (sometimes placed in C. cinerea as C. cinerea subsp. haseltoniana) and C. cinerea including three subspecies (subsp. cinerea, subsp. krainziana and subsp. columna-alba), one of which is often recognised at species level, i.e. the narrow endemic C. krainziana. Here, we evaluate the taxon boundaries of Copiapoa subsection Cinerei using chloroplast sequences and microsatellite data. We generated sequences of three cpDNA markers (rpl32–trnL, trnH–psbA, ycf1) and as indicated in a previous study, found variation between C. gigantea and C. cinerea on a subsample of 34 individuals. Five microsatellite loci were genotyped for 68 individuals from the known range of Copiapoa subsection Cinerei. In contrast with expectations, we found relatively high levels of genetic diversity (e.g., He = 0.775 – 0.827; Ho = 0.580 – 0.750) and no population structure, even between the two species. Additionally, species distribution models were conducted based on abiotic suitability and transformed to 3D maps to account for topographical complexity. The species distribution models and their 3D projections support an allopatric distribution of the four taxa of Copiapoa subsection Cinerei, with each taxon related to a different range with complex topographical features. The obtained molecular results, combined with the presented species distribution modelling, and calculations of extent of occurrence and area of occupancy for the four taxa of Copiapoa subsection Cinerei, suggest a high extinction risk for most of the taxa. A taxonomic treatment is provided.

Members of the cactus family are keystone species of arid and semiarid biomes in the Americas, as they provide shelter and resources to support other members of ecosystems. Extraordinary examples are the several species of flies of the genus Drosophila that lay eggs and feed in their rotting stems, which provide a model system for studying evolutionary processes. Although there is significant progress in understanding the evolution of Drosophila species, there are gaps in our knowledge about the cactus lineages hosting them. Here, we review the current knowledge about the evolution of Cactaceae, focusing on phylogenetic relationships and trends revealed by the study of DNA sequence data. During the last several decades, the availability of molecular phylogenies has considerably increased our understanding of the relationships, biogeography, and evolution of traits in the family. Remarkably, although succulent cacti have very low growth rates and long generation times, they underwent some of the fastest diversifications observed in the plant kingdom, possibly fostered by strong ecological interactions. We have a better understanding of the reproductive biology, population structure and speciation mechanisms in different clades. The recent publication of complete genomes for some species has revealed the importance of phenomena such as incomplete lineage sorting. Hybridization and polyploidization are common in the family, and have been studied using a variety of phylogenetic methods. We discuss potential future avenues for research in Cactaceae, emphasizing the need of a concerted effort among scientists in the Americas, together with the analyses of data from novel sequencing techniques.

The worldwide human population suffering from allergies continues to increase. Pollen grains are a major source of airborne allergens and significant cause of these diseases. Therefore, continuous monitoring of pollen grains released and transported in the air locally or regionally is required to determine the prevalence of various pollen types and identify intra-day and intra-annual seasonal variations over time. In this study, we developed the first pollen calendar for Mexico City, which includes a large variety of taxa, many of which show a long Main Pollen Season which may last throughout the year. The analysis and comparison of daily, monthly and annual values showed that the occurrence and abundance of the main types of aero-allergenic pollen in the atmosphere were species of Fraxinus, Cupressaceae and Alnus, which occur during the periods from December through March, whereas airborne pollens of several species of Poaceae and Urticaceae occurred throughout the year. The variation in pollen concentration showed that the greatest intra-diurnal variations occurred during the second half of the day. Regarding the relationship of pollen with bioclimatic factors, the increase in temperature favoured the presence of pollen in the air, whereas the increase in pluvial precipitation and relative humidity was associated with a decrease in airborne pollen. Large tracts of the Valley of Mexico have atmospheric conditions that are conducive to the accumulation of airborne particles, including pollen. Anomalous winds from the southeast dominated the surface wind variability during the first months of 2010. These patterns induced extreme values in wind convergence at the lower levels of the atmosphere, which resulted in high concentrations of pollen at our sampling site. We suggest that these conditions are related to the warm phase of the El Nino Southern Oscillation phenomenon (2009–2010).

Clinical case The case concerns a 34 year-old man, who presented with pain in the medial canthus in his left eye, with proptosis and diplopia. The examination showed exophthalmus and a palpable mass at the inner canthus. The computed tomography revealed a lesion, which was confirmed by biopsy to be a ossifying fibroma. Discussion Ossifying fibroma is a benign fibro-osseous lesion that mostly affects the jaw. Clinical presentation and complications vary according to its location. Clinical examination, imaging, and histopathology are essential for definitive diagnosis. Surgical treatment is multidisciplinary and long-term follow up is needed.

Vibrio vulnificus, a bacterial species that inhabits brackish waters, is an opportunistic pathogen of humans. V. vulnificus infections can cause acute gastroenteritis, invasive septicemia, tissue necrosis, and potentially death. Virulence factors associated with V. vulnificus include the capsular polysaccharide (CPS), lipopolysaccharide, flagellum, pili, and outer membrane vesicles (OMVs). The aims of this study were to characterize the morphology of V. vulnificus cells and the formation and arrangement of OMVs using cryo-electron microscopy (cryo-EM). cryo-EM and cryo-electron tomography imaging of V. vulnificus strains grown in liquid cultures revealed the presence of OMVs (diameters of ∼45 nm for wild-type, ∼30 nm for the unencapsulated mutant, and ∼50 nm for the non-motile mutant) in log-phase growth. Production of OMVs in the stationary growth phase was limited and irregular. The spacing of the OMVs around the wild-type cells was in regular, concentric rings. In wild-type cells and a non-motile mutant, the spacing between the cell envelope and the first ring of OMVs was ∼200 nm; this spacing was maintained between subsequent OMV layers. The size, arrangement, and spacing of OMVs in an unencapsulated mutant was irregular and indicated that the polysaccharide chains of the capsule regulate aspects of OMV production and order. Together, our results revealed the distinctive organization of V. vulnificus OMVs that is affected by expression of the CPS.

Phylogenomic approaches offer a wealth of data, but a bewildering diversity of methodological choices. These choices can strongly affect the resulting topologies. Here, we explore two controversial approaches (binning genes into "supergenes" and inclusion of only rapidly evolving sites), using new data from hyloid frogs. Hyloid frogs encompass ∼53% of frog species, including true toads (Bufonidae), glassfrogs (Centrolenidae), poison frogs (Dendrobatidae), and treefrogs (Hylidae). Many hyloid families are well-established, but relationships among these families have remained difficult to resolve. We generated a dataset of ultraconserved elements (UCEs) for 50 ingroup species, including 18 of 19 hyloid families and up to 2214 loci spanning >800,000 aligned base pairs. We evaluated these two general approaches (binning, rapid sites only) based primarily on their ability to recover and strongly support well-established clades. Data were analyzed using concatenated likelihood and coalescent species-tree methods (NJst, ASTRAL). Binning strongly affected inferred relationships, whereas use of only rapidly evolving sites did not (indicating ∼87% of the data contributed little information). The optimal approaches for maximizing recovery and support of well-established clades were concatenated likelihood analysis and the use of a limited number of naive bins (statistical binning gave more problematic results). These two optimal approaches converged on similar relationships among hyloid families, and resolved them with generally strong support. The relationships found were very different from most previous estimates of hyloid phylogeny, and a new classification is proposed. The new phylogeny also suggests an intriguing biogeographical scenario, in which hyloids originated in southern South America before radiating throughout the world.

Respiratory syncytial virus (RSV), a member of the Paramyxoviridae family of nonsegmented, negative-sense, single-stranded RNA genome viruses, is a leading cause of lower respiratory tract infections in infants, young children, and the elderly or immunocompromised. There are many open questions regarding the processes that regulate human RSV (hRSV) assembly and budding. Here, using cryo-electron tomography, we identified virus particles that were spherical, filamentous, and asymmetric in structure, all within the same virus preparation. The three particle morphologies maintained a similar organization of the surface glycoproteins, matrix protein (M), M2-1, and the ribonucleoprotein (RNP). RNP filaments were traced in three dimensions (3D), and their total length was calculated. The measurements revealed the inclusion of multiple full-length genome copies per particle. RNP was associated with the membrane whenever the M layer was present. The amount of M coverage ranged from 24% to 86% in the different morphologies. Using fluorescence light microscopy (fLM), direct stochastic optical reconstruction microscopy (dSTORM), and a proximity ligation assay (PLA), we provide evidence illustrating that M2-1 is located between RNP and M in isolated viral particles. In addition, regular spacing of the M2-1 densities was resolved when hRSV viruses were imaged using Zernike phase contrast (ZPC) cryo-electron tomography. Our studies provide a more complete characterization of the hRSV virion structure and substantiation that M and M2-1 regulate virus organization. IMPORTANCE hRSV is a leading cause of lower respiratory tract infections in infants and young children as well as elderly or immunocompromised individuals. We used cryo-electron tomography and Zernike phase contrast cryo-electron tomography to visualize populations of purified hRSV in 3D. We observed the three distinct morphologies, spherical, filamentous, and asymmetric, which maintained comparable organizational profiles. Depending on the virus morphology examined, the amount of M ranged from 24% to 86%. We complemented the cryo-imaging studies with fluorescence microscopy, dSTORM, and a proximity ligation assay to provide additional evidence that M2-1 is incorporated into viral particles and is positioned between M and RNP. The results highlight the impact of M and M2-1 on the regulation of hRSV organization.

The taxonomy of Chilean Cactaceae is notoriously difficult (see e.g., Duarte et al. 2014). Molecular data, and phylogenetic approaches, provided answer to better understand the natural classification of cacti (see e.g., Nyffeler 2002, Edwards 2005, Nyffeler & Eggli 2010, Ritz et al. 2012, Schlumpberger & Renner 2012, Larridon et al. 2015, 2018). As part of an ongoing revision of the Chilean Cacti, which is part of the “Modern Flora of Chile” initiative, nomenclatural novelties are needed in Eulychnia Philippi (1860: 23), Leucostele Backeberg (1953: 36) and Maihueniopsis Spegazzini (1925: 86) (Cactaceae Juss.).

Electron microscopy (EM), cryo-electron microscopy (cryo-EM), and cryo-electron tomography (cryo-ET) are essential techniques used for characterizing basic virus morphology and determining the three-dimensional structure of viruses. Enveloped viruses, which contain an outer lipoprotein coat, constitute the largest group of pathogenic viruses to humans. The purification of enveloped viruses from cell culture presents certain challenges. Specifically, the inclusion of host-membrane derived vesicles, the complete destruction of the viruses, and the disruption of the internal architecture of individual virus particles. Here, we present a strategy for capturing enveloped viruses on affinity grids for use in both conventional EM and cryo-EM/ET applications. We examined the utility of affinity grids for the selective capture of human immunodeficiency virus (HIV) virus-like particles (VLPs), influenza A, and measles virus (MeV). We applied Nickel-nitrilotriacetic acid (Ni-NTA) lipid layers in combination with molecular adaptors to selectively adhere the viruses to the affinity grid surface. This further development of the affinity grid method may prove essential for the gentle and selective purification of enveloped viruses directly onto EM grids for ultrastructural analysis.

Characterization of Human Respiratory Syncytial Virus (HRSV) protein interactions with host cell components is crucial to devise antiviral strategies. Viral nucleoprotein, phosphoprotein and matrix protein genes were optimized for human codon usage and cloned into expression vectors. HEK-293T cells were transfected with these vectors, viral proteins were immunoprecipitated, and co-immunoprecipitated cellular proteins were identified through mass spectrometry. Cell proteins identified with higher confidence scores were probed in the immunoprecipitation using specific antibodies. The results indicate that nucleoprotein interacts with arginine methyl-transferase, methylosome protein and Hsp70. Phosphoprotein interacts with Hsp70 and tropomysin, and matrix with tropomysin and nucleophosmin. Additionally, we performed immunoprecipitation of these cellular proteins in cells infected with HRSV, followed by detection of co-immunoprecipitated viral proteins. The results indicate that these interactions also occur in the context of viral infection, and their potential contribution for a HRSV replication model is discussed.

The assembly of regional biotas and organismal responses to anthropogenic climate change both depend on the capacity of organisms to adapt to novel ecological conditions. Here we demonstrate the concept of evolutionary lag time, the time between when a climatic regime or habitat develops in a region and when it is colonized by a given clade. We analyzed the time of colonization of four clades (three plant genera and one lizard genus) into the Atacama–Sechura Desert of South America, one of Earth’s driest and oldest deserts. We reconstructed time-calibrated phylogenies for each clade and analyzed the timing of shifts in climatic distributions and biogeography and compared these estimates to independent geological estimates of the time of origin of these deserts. Chaetanthera and Malesherbia (plants) and Liolaemus (animal) invaded arid regions of the Atacama–Sechura Desert in the last 10 million years, some 20 million years after the initial onset of aridity in the region. There are also major lag times between when these clades colonized the region and when they invaded arid habitats within the region (typically 4–14 million years). Similarly, hyperarid climates developed ∼8 million years ago, but the most diverse plant clade in these habitats ( Nolana ) only colonized them ∼2 million years ago. Similar evolutionary lag times may occur in other organisms and habitats, but these results are important in suggesting that many lineages may require very long time scales to adapt to modern desertification and climatic change.

Since 2004, governments and non-governmental organizations, together with the fishing communities from nine countries, from Mexico to Peru, have implemented joint efforts to reduce incidental mortality of sea turtles in artisanal longline fisheries of the Eastern Pacific Ocean (EPO). These countries are involved in a Regional Sea Turtle Bycatch Program to achieve this goal. Circle hooks have been proposed as a way to mitigate incidental mortality of sea turtles. Thus, we analyze the performance of circle hooks in relation to J-style and tuna hooks on the hooking rates of target and non-target species in the artisanal surface longline fisheries of three of the participating countries with the largest sample sizes (Ecuador, Panama and Costa Rica). These fisheries target mahi-mahi, Coryphaena hippurus, or a combination of tunas, billfishes and sharks (TBS), and use different techniques and gear configurations to catch their targets. For the TBS fishery we presented the results of comparisons between tuna hooks and 16/0 circle hooks from Ecuador, Panama and Costa Rica, and between tuna hooks and 18/0 circle hooks in Costa Rica. For the mahi-mahi fishery, we analyzed the performance of 14/0 and 15/0 circle hooks in Ecuadorian vessels and 16/0 circle hooks in Costa Rican vessels vs. the traditional J-style hooks. A total of 730,362 hooks were observed in 3126 sets. Hooking rates for target and non-target species were not consistent for all fisheries and countries analyzed. However, circle hooks reduced sea turtle hooking rates in most of the comparisons.

Bacteriophage particles contain both simple and complex macromolecular assemblages and machines that enable them to regulate the infection process under diverse environmental conditions with a broad range of bacterial hosts. Recent developments in cryo-electron tomography (cryo-ET) make it possible to observe the interactions of bacteriophages with their host cells under native-state conditions at unprecedented resolution and in three-dimensions. This review describes the application of cryo-ET to studies of bacteriophage attachment, genome ejection, assembly and egress. Current topics of investigation and future directions in the field are also discussed.

We report the largest study on the prevalence and distribution of HCV genotypes in Spain (2000-2015), and we relate them with clinical, epidemiological and virological factors. Patients from 29 hospitals in 10 autonomous communities (Andalusia, Aragon, Castilla-Leon, Catalonia, Galicia, Canary Islands, Madrid Community, Valencian Community, Murcia Region and Basque Country) have been studied. Annual distribution of HCV genotypes and subtypes, as well as gender, age, transmission route, HIV and/or HBV coinfection, and treatment details were recorded. We included 48595 chronically HCV-infected patients with the following characteristics: median age 51 years (IQR, 44-58), 67.9% male, 19.1% HIV-coinfected, 23.5% HBV-coinfected. Parenteral transmission route was the most frequent (58.7%). Genotype distribution was 66.9% GT1 (24.9% subtype 1a and 37.9% subtype 1b), 2.8% GT2, 17.3% GT3, 11.4% GT4 and 0.1% GT5 and 0.02% GT6. LiPA was the most widely HCV genotyping test used (52.4%). HCV subtype 1a and genotypes 3 and 4 were closely associated with male gender, parenteral route of infection and HIV and HBV coinfection; in contrast, subtype 1b and genotype 2 were associated with female gender, nonparenteral route and mono-infection. Age was related to genotype distribution, and different patterns of distribution and biodiversity index were observed between different geographical areas. Finally, we describe how treatment and changes in transmission routes may have affected HCV genotype prevalence and distribution patterns. We present the most recent data on molecular epidemiology of hepatitis C virus in Spain. This study confirms that genotype distributions vary with age, sex, HIV and HBV coinfection and within geographical areas and epidemiological groups.

Locust is one of the most predominant agricultural pests in Mexico, followed by non-migratory grasshoppers. Their devastating effects cover numerous Mexican states, generating important economic losses. Metarhizium acridum (Hypocreales: Clavicipitaceae) has been employed in Mexico as a microbial agent for biological control for some time. However, questions related to the long-term persistence and viability of the fungus after it has been sprayed over crop fields remain to be elucidated. To resolve these questions, we performed a study in a crop field where an abundant population of the grasshopper Sphenarium purpurascens (Orthoptera: Pyrgomorphidae) is present. The fungus M. acridum was sprayed during August and October 2007. Its presence in the air, soil and vegetative cover was subsequently monitored using molecular methods and culture techniques. The results showed that the fungus was viable on the ground during the follow-up period (66 weeks). Its population fluctuated markedly, however, reaching four times its initial concentration and later decreasing to nearly half its initial concentration by the end of the study. The initial concentrations of the fungus in the air and in the vegetative cover decayed steadily (with some small, isolated recoveries) until its complete disappearance after 8 months.

Background: The Cactaceae is a diverse and conspicuous Neotropical family that has evolved a wide variety of adaptations during co-evolution with their interacting species. Recent research has indicated complex ecological and evolutionary interactions involving cacti and other organisms. Aims: We reviewed four studies involving cacti that have important implications for our understanding of the evolution of life traits and maintenance of cactus diversity. Also, these studies illustrate how the modern theoretical background of the ecology and evolution of species interactions is influencing the research in Cactaceae. Methods: The studies showed here are (1) the evolutionary ecology of a mistletoe-cactus parasitism in central Chile, (2) the effect of an exotic grass on the demography of a threatened cactus in Puerto Rico, (3) the herpetochory in a tropical genus of cacti in Venezuela, and (4) the role of abiotic and biotic factors on the floral morphology in globose cacti species in northern Chile. Results:...

We describe a technique to detect the presence of airborne conidia from the fungus M.acridum (formerly Metarhizium anisopliae var. acridum) (Hypocreales: Clavicipitaceae) with great accuracy. Airborne conidia were collected using Hirst-type spore traps. DNA extractions were optimized to achieve the best possible recovery. DNA was examined using polymerase chain reaction (PCR) with specific oligonucleotides to enable the detection of a single conidium. Experiments using a mini-wind tunnel were conducted to validate the method. Subsequently, this technique was applied to an agricultural region of Mexico, where M. acridum was sprayed to control the grasshopper, Sphenarium purpurascens, population (Orthoptera: Pyrgomorphidae). M. acridum conidia were detected 2 days after spraying in San Mateo Coatepec (Puebla, site of grasshopper study).

The classification of Eriosyce subgenus Neoporteria (“subsection” in the sense of Kattermann) and the role of allopatry/sympatry in the diversification of the group were studied by use of cladistic and predictive distribution modeling methods. We reconstructed the phylogenetic relationships of subgenus Neoporteria by analyzing 38 morphological characters and DNA sequences from two chloroplast regions of 21 taxa from the Chilean subsections of Eriosyce using a Bayesian and maximum likelihood phylogenetic framework. Also, we attempted to find out if the divergence between the sister taxa in the Neoporteria group had been caused by allopatric or sympatric mechanisms. The morphology-based analysis placed E. chilensis basal within the Neoporteria clade and suggested a further broadening of the group by including E. taltalensis var. taltalensis, formerly considered a member of subsection Horridocactus. However, the combined DNA data placed E. sociabilis and E. taltalensis var. taltalensis within the Horridocactus clade, and placed E. chilensis with E. subgibbosa var. litoralis. The broad concept of E. subgibbosa sensu Kattermann (comprising seven infraspecific taxa), was rejected by our combined molecular results. Finally, our results corroborated changes in subsection Neoporteria proposed by various authors and suggested further modifications within Neoporteria. The analyses of the degree of geographic overlap of the predicted distributions indicated null overlap between the sister taxa, and one probable hybrid origin of E. chilensis, indicating that evolutionary divergence is mainly caused by an allopatric process associated with climatic tolerance.