Breeding for Persistence in Lotus Corniculatus

The objectives of this study were to develop a more persistent germplasm of birdsfoot trefoil (Lotus corniculatus L.) and to evaluate its potential use in Uruguay. Two cycles of phenotypic recurrent selection were done under field conditions. Eight hundred and 1600 spaced plants were established for the first and second cycle of selection. Both nurseries were visually assessed for crown rot, relative growth, plant habit and foliar diseases. The final selection of the breeding population LE 65-56 includes thirty six plants selected from sixteen elite families. This selection was more persistent and productive than the check varieties in plot evaluation. On average, LE 65-56 produced 12%, 16% and 114% more forage than cv. San Gabriel in the first, second and third year, respectively. Selecting for crown health and relative growth in the spaced plant nurseries increased persistence and forage yields in plots under cut

Overwintering of Piezodorus guildinii (Heteroptera, Pentatomidae) populations.

Piezodorus guildinii (Westwood) is a soybean pest that causes significant economic losses in the Americas. The variability of overwintering (diapause) traits was evaluated in populations of the Southwest (SW) (33°55′?34°17′S, 57°13′?57°46′W) during 2-year period (2011?2013) and of the Northwest (NW) (32°01′?33°02′S, 57°50′?57°24′W) during 1-year period (2014?2015) Regions of Uruguay. Samples were taken from different plant species (cultivated legumes, wild shrubs, and trees) and from overwintering sites (leaf litter and bark). Alfalfa, Medicago sativa L. was the main host, with a collection period of 10?11 months in the SW and 12 months in the NW. Cluster analysis for each sex was carried out to group the months according to the similarity in diapause traits of populations (body size, body lipid content, immature reproductive organs, and clear type of pronotum band and connexivum in females). Female diapause in the SWwas longer (beginning of autumn to end of winter) than that in the NW (mid-autumn to mid-winter). Male diapause was longer (mid-autumn o mid-winter) in SW1 (1st year) than in SW2 (2nd year) and NW (lateautumn to mid-winter). In both regions, male diapause was shorter than female. Differences were associated with maximum temperature at daylight hours ≤ 12.1, being necessary maximum temperatures below 23.8 °C for females and 19.2 °C for males to initiate diapause

Accumulation of Mg to Diffusive Gradients in Thin Films (DGT) devices: Kinetic and thermodynamic effects of the ionic strength

Availability of magnesium is a matter of concern due to its role in many environmental and biological processes. Diffusive Gradients in Thin Films (DGT) devices can measure Mg availability in situ. This work shows that Mg accumulation in water largely increases when ionic strength (I) decreases. This phenomenon can be explained from (i) the increase of both the association equilibrium (K) and rate (ka,R) constants for the reaction between Mg cations and resin sites, and (ii) the growing contribution of the partitioning of Mg cations at the resin–gel interface, as I decreases. Two theoretical models that take into account electrical interactions among Mg cations, background electrolyte, and resin sites can successfully be used to determine ka,R and K at each I. Both models yield similar ka,R values, which fulfill an expression for the kinetic salt effect. For freshwater (with a typical salinity of 10 mM and circumneutral pH), the binding of Mg is so fast and strong that the simplest perfect-sink DGT expression can be helpful to predict (overestimation lower than 5%) the accumulation in solutions with Mg concentrations up to 1 mM whenever the deployment time is below 9 h. Perfect sink conditions can still be applied for longer times, in systems with either a lower I or a lower Mg concentration.Financial support from FEDER and the Spanish Ministry of Education and Science (Projects CTM2012-39183 and CTM2013-48967) is gratefully acknowledged

Salmonella prevalence serotypes and patterns of antimicrobial resistance in cohorts of nursery and finishing pigs

Salmonella are now being monitored by the Food Safety Inspection Service of the USDA for purposes of process control in slaughter plants, and as a sentinel organism for emerging antimicrobial resistance (I). Public concerns about the emergence of foodborne bacteria with multi-resistant phenotypes, and about the use of antimicrobials in food animals, are increasing. Despite considerable research, the epidemiology of asymptomatic Salmonella in pigs is poorly understood (2). The purpose of this paper is to examine the association between nursery prevalence of Salmonella and finisher prevalence, and to describe patterns of antimicrobial use and antimicrobial resistant Salmonella in modern swine rearing facilities

Determinants of the voltage dependence of G protein modulation within calcium channel β subunits

CaVβ subunits of voltage-gated calcium channels contain two conserved domains, a src-homology-3 (SH3) domain and a guanylate kinase-like (GK) domain with an intervening HOOK domain. We have shown in a previous study that, although Gβγ-mediated inhibitory modulation of CaV2.2 channels did not require the interaction of a CaVβ subunit with the CaVα1 subunit, when such interaction was prevented by a mutation in the α1 subunit, G protein modulation could not be removed by a large depolarization and showed voltage-independent properties (Leroy et al., J Neurosci 25:6984–6996, 2005). In this study, we have investigated the ability of mutant and truncated CaVβ subunits to support voltage-dependent G protein modulation in order to determine the minimal domain of the CaVβ subunit that is required for this process. We have coexpressed the CaVβ subunit constructs with CaV2.2 and α2δ-2, studied modulation by the activation of the dopamine D2 receptor, and also examined basal tonic modulation. Our main finding is that the CaVβ subunit GK domains, from either β1b or β2, are sufficient to restore voltage dependence to G protein modulation. We also found that the removal of the variable HOOK region from β2a promotes tonic voltage-dependent G protein modulation. We propose that the absence of the HOOK region enhances Gβγ binding affinity, leading to greater tonic modulation by basal levels of Gβγ. This tonic modulation requires the presence of an SH3 domain, as tonic modulation is not supported by any of the CaVβ subunit GK domains alone

Performance of Lotus corniculatus L. genotypes submitted to cutting interval: subsidies to a breeding program

The objective of this work was to evaluate the response of five birdsfoot trefoil populations (Lotus corniculatus L.), selected under severe and intense grazing (P38, P37, P9 and P3) or cutting (population Corte), one rhizomatous population from Morocco and two cultivars, São Gabriel (Brazil) and ARS 2620 (USA), to different cutting intervals (20 and 40 days). The trial was carried out in a greenhouse for 224 days. The plants were submitted to the treatments during four months, when it was made four and two cuttings for the 20 and 40 day intervals, respectively. In the following months, aerial part of the plants was totally removed and after 70 days of regrowth, evaluation of roots and aerial section were performed. Analysis of variance and multivariate analysis was performed, obtaining the Mahalanobis distance (Md), dendogram by UPGMA method and the relative contribution of the characters for genetic divergence. There was no genotype × cutting interval interaction and the genotypes presented a reduction on dry matter yield of roots, crown and aerial sections, number of stems and plant height when submitted to frequent cuttings. The most divergent genotypes were Marrocos and P9 (Md = 108.7) and the most similar ones were São Gabriel and P37 (Md = 7.8). The results suggest exclusion of the population P9 because of its weak performance and the utilization of the populations Corte and P37 as progenitors in the birdsfoot trefoil breeding program. Root dry matter accumulation and plant height were the characters that contributed most to genetic divergence and they can be used for selection works

Engineering the Salmonella type III secretion system to export spider silk monomers

The type III secretion system (T3SS) exports proteins from the cytoplasm, through both the inner and outer membranes, to the external environment. Here, a system is constructed to harness the T3SS encoded within Salmonella Pathogeneity Island 1 to export proteins of biotechnological interest. The system is composed of an operon containing the target protein fused to an N-terminal secretion tag and its cognate chaperone. Transcription is controlled by a genetic circuit that only turns on when the cell is actively secreting protein. The system is refined using a small human protein (DH domain) and demonstrated by exporting three silk monomers (ADF-1, -2, and -3), representative of different types of spider silk. Synthetic genes encoding silk monomers were designed to enhance genetic stability and codon usage, constructed by automated DNA synthesis, and cloned into the secretion control system. Secretion rates up to 1.8 mg l−1 h−1 are demonstrated with up to 14% of expressed protein secreted. This work introduces new parts to control protein secretion in Gram-negative bacteria, which will be broadly applicable to problems in biotechnology

Production of microsclerotia from entomopathogenic fungi and use in maize seed coating as delivery for biocontrol against Fusarium graminearum

The commercial use of the entomopathogenic fungi Metarhizium spp. in biopesticides has gained more interest since the discovery that several species of this genus are able to colonize roots. In general, commercial products with Metarhizium are formulated based on conidia for insect pest control. The process of mass production, harvesting, and formulation of infective conidia can be detrimental for conidial viability. Entomopathogenic fungi such as Metarhizium spp. are able to produce high concentrations of resistant structures, known as microsclerotia, when grown in liquid media. Microsclerotia are desiccation tolerant, with excellent storage stability, and are capable of producing high quantities of infective conidia after rehydration. The aim of this study was to evaluate microsclerotia production by different isolates of Metarhizium spp. and determine the effect of microsclerotia coated onto maize seeds on plant growth in the presence of soil-borne pathogen Fusarium graminearum. On average, ~1 × 10⁵ microsclerotia/mL were produced by selected isolates of M. anisopliae (A1080 and F672) and Metarhizium robertsii (F447). Microsclerotia were formulated as granules with diatomaceous earth and used for seed coating, after which propagules produced around 5 × 10⁶ CFU/g of seeds. In the presence of the plant pathogen, maize plants grown from untreated seeds had the lowest growth, while plants treated with the Metarhizium microsclerotia had significantly greater growth than the control plants. Hyphae were observed growing on and in root tissues in all the Metarhizium spp. treatments but not in samples from control plants. Metarhizium hyphal penetration points' on roots were observed 1 month after sowing, indicating the fungi were colonizing roots as endophytes. The results obtained indicate that microsclerotia can be coated onto seeds, providing plant protection against soil plant pathogens and a method to establish Metarhizium in the ecto- and endo-rhizosphere of maize roots, allowing the persistence of this biocontrol agent

Characterization of the role of ribonucleases in Salmonella small RNA decay

In pathogenic bacteria, a large number of sRNAs coordinate adaptation to stress and expression of virulence genes. To better understand the turnover of regulatory sRNAs in the model pathogen, Salmonella typhimurium, we have constructed mutants for several ribonucleases (RNase E, RNase G, RNase III, PNPase) and Poly(A) Polymerase I. The expression profiles of four sRNAs conserved among many enterobacteria, CsrB, CsrC, MicA and SraL, were analysed and the processing and stability of these sRNAs was studied in the constructed strains. The degradosome was a common feature involved in the turnover of these four sRNAs. PAPI-mediated polyadenylation was the major factor governing SraL degradation. RNase III was revealed to strongly affect MicA decay. PNPase was shown to be important in the decay of these four sRNAs. The stability of CsrB and CsrC seemed to be independent of the RNA chaperone, Hfq, whereas the decay of SraL and MicA was Hfq-dependent. Taken together, the results of this study provide initial insight into the mechanisms of sRNA decay in Salmonella, and indicate specific contributions of the RNA decay machinery components to the turnover of individual sRNAs

BarA-UvrY Two-Component System Regulates Virulence of Uropathogenic E. coli CFT073

Uropathogenic Escherichia coli (UPEC), a member of extraintestinal pathogenic E. coli, cause ∼80% of community-acquired urinary tract infections (UTI) in humans. UPEC initiates its colonization in epithelial cells lining the urinary tract with a complicated life cycle, replicating and persisting in intracellular and extracellular niches. Consequently, UPEC causes cystitis and more severe form of pyelonephritis. To further understand the virulence characteristics of UPEC, we investigated the roles of BarA-UvrY two-component system (TCS) in regulating UPEC virulence. Our results showed that mutation of BarA-UvrY TCS significantly decreased the virulence of UPEC CFT073, as assessed by mouse urinary tract infection, chicken embryo killing assay, and cytotoxicity assay on human kidney and uroepithelial cell lines. Furthermore, mutation of either barA or uvrY gene reduced the production of hemolysin, lipopolysaccharide (LPS), proinflammatory cytokines (TNF-α and IL-6) and chemokine (IL-8). The virulence phenotype was restored similar to that of wild-type by complementation of either barA or uvrY gene in trans. In addition, we discussed a possible link between the BarA-UvrY TCS and CsrA in positively and negatively controlling virulence in UPEC. Overall, this study provides the evidences for BarA-UvrY TCS regulates the virulence of UPEC CFT073 and may point to mechanisms by which virulence regulations are observed in different ways may control the long-term survival of UPEC in the urinary tract