Assessing stomatal response to live bacterial cells using whole leaf imaging.

Stomata are natural openings in the plant epidermis responsible for gas exchange between plant interior and environment. They are formed by a pair of guard cells, which are able to close the stomatal pore in response to a number of external factors including light intensity, carbon dioxide concentration, and relative humidity (RH). The stomatal pore is also the main route for pathogen entry into leaves, a crucial step for disease development. Recent studies have unveiled that closure of the pore is effective in minimizing bacterial disease development in Arabidopsis plants; an integral part of plant innate immunity. Previously, we have used epidermal peels to assess stomatal response to live bacteria (Melotto et al. 2006); however maintaining favorable environmental conditions for both plant epidermal peels and bacterial cells has been challenging. Leaf epidermis can be kept alive and healthy with MES buffer (10 mM KCl, 25 mM MES-KOH, pH 6.15) for electrophysiological experiments of guard cells. However, this buffer is not appropriate for obtaining bacterial suspension. On the other hand, bacterial cells can be kept alive in water which is not proper to maintain epidermal peels for long period of times. When an epidermal peel floats on water, the cells in the peel that are exposed to air dry within 4 hours limiting the timing to conduct the experiment. An ideal method for assessing the effect of a particular stimulus on guard cells should present minimal interference to stomatal physiology and to the natural environment of the plant as much as possible. We, therefore, developed a new method to assess stomatal response to live bacteria in which leaf wounding and manipulation is greatly minimized aiming to provide an easily reproducible and reliable stomatal assay. The protocol is based on staining of intact leaf with propidium iodide (PI), incubation of staining leaf with bacterial suspension, and observation of leaves under laser scanning confocal microscope. Finally, this method allows for the observation of the same live leaf sample over extended periods of time using conditions that closely mimic the natural conditions under which plants are attacked by pathogens

Efficacy Assessment of Timber Based In-Plane Strengthening of Wooden Floors on the Seismic Response of Masonry Structures by means of DEM Analyses

Masonry buildings are highly vulnerable to seismic loading, and their dynamic response is strongly influenced by the timber floor in-plane deformability and by the quality of the wall-to-floor connections. Understanding the behavior of timber floors and roofs and their interaction with the masonry walls is therefore important for the protection of historical buildings. In a previous research project, different timber-based dry-connected floor strengthening solutions were tested under in-plane loads. The experimental results show a significant increase in shear strength and stiffness. Discrete Element Method is here used to evaluate the effectiveness of the strengthening solutions in avoiding the triggering of the out-of-plane collapse of masonry walls, first on a simple masonry cell, and then on a heritage listed masonry building. A detailed cyclic model of the floor behavior was implemented: the unreinforced and reinforced floors were described by beams connected with nonlinear springs, reproducing the experimental hysteretic response. Both the case studies highlight the effectiveness of the strengthening solutions in reducing the out-of-plane displacements of masonry walls, confirmed also by a comparison with the ideal rigid diaphragm case. The reinforced floor is able to transfer the seismic forces to the shear-resistant walls. The out-of-plane displacements are compatible with the wall capacity, and the reinforced floor hysteretic cycles contribute to dissipate part of the input energy. Moreover, a proper connection design can also cap the transferred seismic forces to an acceptable level for shear-resistant walls

Evaluation of timber floor in-plane retrofitting interventions on the seismic response of masonry structures by DEM analysis: a case study

The seismic response of existing masonry structures is strongly influenced by floor and roof in-plane properties. A strengthening intervention is often needed for traditional timber floors to overcome their low in-plane stiffness and to preserve historical buildings. In this study, the effects of unreinforced and reinforced timber floors on the seismic behaviour of an existing listed masonry building are investigated with dynamic non-linear analyses by means of the Discrete Element Method (DEM). With this approach, the failure processes and collapse sequences of masonry structures can be captured in detail. A previously developed model of the floor cyclic behaviour, based on experimental data, is applied herein to DEM models of the masonry building. Different seismic ground accelerations, different floor types and different floor-to-wall connections are considered. The results highlight the effectiveness of the analysed floor strengthening solution in reducing the out-of-plane displacements of masonry walls. With adequate connections, the reinforced floor is able to transfer the seismic forces to the shear-resistant walls up to the shear-sliding collapse of the structural sidewalls. A comparison with the ideal rigid diaphragm case confirms the good performance of the strengthened floors. The small observed out-of-plane displacements are compatible with the masonry wall capacity, and the reinforced floor hysteretic cycles contribute to dissipate part of the input energy. Moreover, different designs of the connections can also cap the transferred seismic forces to an acceptable level for shear-resistant walls

La diversidad (in)visible. Identidad(es) entre policías bonaerenses

En este artículo analizamos cómo se construye el “nosotros” policial, entendiendo que el mismo no responde a una construcción estable y perenne propia de una esencia ontológica invariable del “ser policial”; sino que, por el contrario, es el resultado voluble y mutante de las múltiples relaciones sociales. Para dar cuenta de esta construcción estudiaremos algunos clivajes internos de la institución policial, exhibiendo la diversidad de vínculos sociales existentes en el interior de una grupalidad que se presenta como homogénea. La imagen del “verdadero policía”, idealización del hacer policial y su representación, nos permitirá rastrear la construcción de una distinción para con la sociedad, al mismo tiempo, que pondrá luz sobre la heterogeneidad opacada

Salmonella enterica Serovar Typhimurium 14028s Genomic Regions Required for Colonization of Lettuce Leaves.

Contamination of edible produce leaves with human bacterial pathogens has been associated with serious disease outbreaks and has become a major public health concern affecting all aspects of the market, from farmers to consumers. While pathogen populations residing on the surface of ready-to-eat produce can be potentially removed through thorough washing, there is no disinfection technology available that effectively eliminates internal bacterial populations. By screening 303 multi-gene deletion (MGD) mutants of Salmonella enterica serovar Typhimurium (STm) 14028s, we were able to identify ten genomic regions that play a role in opening the stomatal pore of lettuce leaves. The major metabolic functions of the deleted regions are associated with sensing the environment, bacterium movement, transport through the bacterial membrane, and biosynthesis of surface appendages. Interestingly, at 21 days post inoculation, seven of these mutants showed increased population titers inside the leaf, two mutants showed similar titers as the wild type bacterium, whereas one mutant with a large deletion that includes the Salmonella pathogenicity island 2 (SPI-2) showed significantly impaired persistence in the leaf apoplast. These findings suggest that not all the genomic regions required for initiation of leaf colonization (i.e., epiphytic behavior and tissue penetration) are essential for continuing bacterial survival as an endophyte. We also observed that mutants lacking either SPI-1 (Mut3) or SPI-2 (Mut9) induce callose deposition levels comparable to those of the wild type STm 14028s; therefore, these islands do not seem to affect this lettuce defense mechanism. However, the growth of Mut9, but not Mut3, was significantly impaired in the leaf apoplastic wash fluid (AWF) suggesting that the STm persistence in the apoplast may be linked to nutrient acquisition capabilities or overall bacterial fitness in this niche, which are dependent on the gene(s) deleted in the Mut9 strain. The genetic basis of STm colonization of leaves investigated in this study provides a foundation from which to develop mitigation tactics to enhance food safety

Coronatine Facilitates Pseudomonas syringae Infection of Arabidopsis Leaves at Night.

In many land plants, the stomatal pore opens during the day and closes during the night. Thus, periods of darkness could be effective in decreasing pathogen penetration into leaves through stomata, the primary sites for infection by many pathogens. Pseudomonas syringae pv. tomato (Pst) DC3000 produces coronatine (COR) and opens stomata, raising an intriguing question as to whether this is a virulence strategy to facilitate bacterial infection at night. In fact, we found that (a) biological concentration of COR is effective in opening dark-closed stomata of Arabidopsis thaliana leaves, (b) the COR defective mutant Pst DC3118 is less effective in infecting Arabidopsis in the dark than under light and this difference in infection is reduced with the wild type bacterium Pst DC3000, and (c) cma, a COR biosynthesis gene, is induced only when the bacterium is in contact with the leaf surface independent of the light conditions. These findings suggest that Pst DC3000 activates virulence factors at the pre-invasive phase of its life cycle to infect plants even when environmental conditions (such as darkness) favor stomatal immunity. This functional attribute of COR may provide epidemiological advantages for COR-producing bacteria on the leaf surface

The (in)visible diversity. Identity (s) among the police of Buenos Aires

En este artículo analizamos cómo se construye el "nosotros" policial, entendiendo que el mismo no responde a una construcción estable y perenne propia de una esencia ontológica invariable del "ser policial"; sino que, por el contrario, es el resultado voluble y mutante de las múltiples relaciones sociales. Para dar cuenta de esta construcción estudiaremos algunos clivajes internos de la institución policial, exhibiendo la diversidad de vínculos sociales existentes en el interior de una grupalidad que se presenta como homogénea. La imagen del "verdadero policía", idealización del hacer policial y su representación, nos permitirá rastrear la construcción de una distinción para con la sociedad, al mismo tiempo, que pondrá luz sobre la heterogeneidad opacada.In this article we analyze how the "we" police is built, understanding that it does not respond to a stable and permanent construction, typical of an invariable ontological essence of "being police", but on the contrary, it is the result of fickle and mutant multiple social relations. To account for this construction we will study some police internal cleavages, showing the diversity of existing social ties within a groupality presented as homogeneous. The image of the "real police", idealized representation of police doing, will allow us to track the construction of a distinction to society, at the same time it will light up the obscured heterogeneity.Fil: Garriga Zucal, Jose Antonio. Universidad Nacional de San Martín. Instituto de Altos Estudios Sociales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Melotto, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Rapid adaptation to invasive predators overwhelms natural gradients of intraspecific variation

Invasive predators can exert strong selection on native populations. If selection is strong enough, populations could lose the phenotypic variation caused by adaptation to heterogeneous environments. We compare frog tadpoles prior to and 14 years following invasion by crayfish. Prior to the invasion, populations differed in their intrinsic developmental rate, with tadpoles from cold areas reaching metamorphosis sooner than those from warm areas. Following the invasion, tadpoles from invaded populations develop faster than those from non-invaded populations. This ontogenetic shift overwhelmed the intraspecific variation between populations in a few generations, to the point where invaded populations develop at a similar rate regardless of climate. Rapid development can have costs, as fast-developing froglets have a smaller body size and poorer jumping performance, but compensatory growth counteracts some costs of development acceleration. Strong selection by invasive species can disrupt local adaptations by dampening intraspecific phenotypic variation, with complex consequences on lifetime fitness

Laurent Fourcaut, Didactique de la fleur. “Angélique”, matrice de l’œuvre gionienne

Pubblicato per la prima volta nel 1989 nella collana «Archives des lettres modernes» e nella serie «Archives Jean Giono» diretta da Michel Minard, il testo che viene ora ristampato presenta uno studio di taglio prevalentemente psicanalitico su Angélique (1980), primo romanzo incompiuto di Giono. Fourcaut individua i segni di una dialettica tra due poli che caratterizzerà l’intera produzione: la perdita, ovvero il desiderio di fondersi nel mondo-madre, e l’avarizia, ovvero l’istinto di conserv..