Caracterización simbiótica y filogenética de rizobios que nodulan la nueva especie Lupinus mariae-josephi

Una nueva especie de altramuz, Lupinus mariae-josephi, ha sido identificado recientemente en Valencia (Pascual, H.). Esta especie, a diferencia de las descritas en la Península ibérica y en el viejo mundo, no crece en suelos ácidos sino en suelos alcalinos y con alto contenido en calcio. El objetivo general de este proyecto es investigar si existen diferencias fenotípicas y genéticas entre los rizobios que nodulan Lupinus mariae-josephi y lupinos de suelos ácidos nativos de la Península Ibérica (Lupinus angustifolius, L.luteus y otras cuatro especies). En este proyecto se han aislado bacterias (rizobios) de nódulos de L. maria-josephi a partir de suelos básicos de Valencia (localidad de LLombai) y se está realizando su caracterización a nivel morfológico, nutricional, simbiótico y molecular. A nivel molecular se han comparado los genes “housekeeping” 16S rRNA, recA, atpD, gln2 y el simbiótico, nodC de diversas cepas que nodulan L. mariae-josephi y con cepas de otras especies de rizobios. Por otra parte, también se está evaluando la capacidad de L. mariae-josephi de ser nodulada por diferentes rizobios bajo condiciones bacteriológicamente controladas

Dissecting the microglial response in transgenic models of amyloidogenesis and tauopathy

Amyloid-beta (Abeta) peptide deposits and hyperphosphorylated tau protein (phospho-tau) accumulate in Alzheimer’s disease (AD) brains. These abnormal protein aggregates leads to glial activation, synaptic dysfunction, neuronal loss and cognitive decline. While microglial response has mostly been analyzed in relation to Abeta accumulation, little is still known about inflammatory processes associated with tau pathology. Microglial reactivity and defective glial responses have been involved in these proteinopathies. Our aim is to clarify the effects of Abeta and tau separately, in order to improve the comprehension of their differential contribution to neuroinflammation and neurodegeneration. We compared the progression of these processes in an amyloidogenic AD model (APPSL/PS1M146L) and two different models of tauopathy (ThyTau22 and hP301S) from 2 to 18 months of age. Accumulation of aggregated proteins was assessed using specific anti- Abeta and phospho-tau antibodies. Inflammatory response was studied using a battery of microglial markers (Iba1, CD45, CD68, Trem2 and Gal-3). In the hippocampus of these models, Tau and Abeta pathologies initiated as early as 2 months of age and increased progressively with aging. Neuritic plaques induced a strong microglial activation associated to plaques in APP/PS1 mice. Interestingly, inflammatory markers and microglial reactivity were barely increased in the hippocampus of ThyTau mice in contrast to not only APP/PS1, but also to P301S mice, which displayed a prominent microglial response. Deciphering the specific effects of Abeta, tau and their different toxic species, would indeed enable the development of novel therapeutic strategies and drugs targeting neuroinflammatory pathways related to these proteinopathies.Universidad de Málaga. Campus de excelencia Andalucía-Tech. Supported by PI18/01557 (AG) and PI18/01556 (JV) grants from ISCiii of Spain co-financed by FEDER funds from European Union, and by grant PPIT.UMA.B1.2017/26 (RS-V)

Phenotypic and phylogenetic characterization of endosymbiotic bacteria from Lupinus mariae-josephi

Lupinus mariae-josephi is a Lupinus species that thrives in a Southeastern area of Spain (Valencia) in soils of singularly high pH and active lime content. It is nodulated by extra-slow growing bacteria symbiotically and phylogenetically distant to endosymbiotic strains nodulating other Lupinus sp. native of the Iberian Peninsula and adapted to growth in acid soils. Cross-inoculation experiments revealed that the L. mariae-josephi endosymbiotic bacteria are unable to nodulate or efficiently fix nitrogen with well-known Lupinus spp. Their species affiliation was examined by a multilocus sequence analysis of four housekeeping genes (16S rDNA, glnII, recA, atpD) and the symbiotic nodC gene. Single and concatenated phylogenetic analyses of these genes consistently revealed that L. mariae-josephi endosymbiotic bacteria belong to a clade, within the Bradyrhizobium genus, highly differentiated from the Bradyrhizobium clade that includes currently named Bradyrhizobium species as well as the endosymbiotic bacteria from Lupinus species tested in this study. Within this new clade the L. mariae-josephi bacteria nested in several subgroup that may correspond to novel sister species. The phylogenetic analysis based on the nodC gene showed that L. mariae-josephi endosymbiotic bacteria define a novel branch of the nodC Bradyrhizobium tree and likely have a common unique ancestor for the symbiotic genes with nodule isolates from Retama spp

Eye-tracker study of the developmental eye movement test in young people without binocular dysfunctions

Background: The purpose of this study was to evaluate ocular motility in normal young adults when performing the Developmental Eye Movement (DEM) test using an infrared eye-tracker in a sample of young subjects without visual dysfunctions. Methods: An optometric evaluation was carried out on 52 participants with a mean age of 21.00 ± 3.22 years to verify they did not have any binocular dysfunction, by completing a computerized version of the DEM test while their eye movements were recorded with an eye-tracker. A custom-written software was developed to analyse some specific parameters of ocular motility while performing each subtest (Test A, Test B and Test C) of the complete DEM test. Results: The mean duration of the fixations was shorter in Test C (243.56 ± 46.18 s) than in Test A (493.52 ± 171.41 s) and Test B (484.20 ± 156.59 s). The mean adjusted horizontal (AdjHT: 35.24 ± 6.68 s) and vertical (VT: 33.58 ± 5.56 s) times were at the 45th and at the 40th percentile, respectively. In Test C, there was a high positive significant correlation between the saccadic speed (cc: 0.77; p < 0.001) and the saccadic length (cc: 0.74; p < 0.001) of both eyes. Conclusions: The eye-tracker is an objective method to evaluate the DEM test in subjects without binocular dysfunctions, measuring and quantifying ocular motility parameters that are impossible with the traditional subjective method. The eye movements of both eyes are conjugated in each subject, having saccades of the same length and speed

Lupinus mariae-josphi, a new lupin endemic of soils with active lime and high pH in South Eastern Spain, is nodulated by a new bacterial lineage within Bradyrhizobium genus

Lupinus mariae-josephi is a recently described species (Pascual, 2004) able to grow in soils with high pH and active lime content in the Valencia province (Spain). L. mariae-josephi endosymbionts are extremely slowgrowing bacteria with genetic and symbiotic characteristics that differentiate them from Bradyrhizobium strains nodulating Lupinus spp. native of the Iberian Peninsula and adapted to grow in acid soils. Cross-inoculation experiments revealed that all the endosymbiotic isolates from L. mariae-josephi tested are legume-host selective and are unable to nodulate species such as L. angustifolius, and L. luteus. In contrast, Bradyrhizobium strains from Lupinus spp. tested were able to nodulate L. mariae-josephi, although the nodules fixed nitrogen inefficiently. Phylogenetic analysis was performed with housekeeping genes (rrn, glnII, recA, atpD) and nodulation gene nodC. Housekeeping gene phylogeny revealed that L. mariae-josephi rhizobia form a strongly supported monophyletic group within Bradyrhizobium genus. This cluster also includes B. jicamae and certain strains of B. elkanii. Contrarily, isolates from other Lupinus spp. native of the Iberian Peninsula were grouped mainly within B. canariense and two B. japonicum lineages. Phylogenetic analysis of L. mariae-josephi isolates based on the nodC symbiotic gene defined a solid clade close to isolates from Algerian Retama spp. and to fast-growing rhizobia

Taxonomy of endosymbiotic bacteria from a novel Lupinus sp. (Lupinus mariae-josephi) endemic of a limed-alkaline soil habitat in Southeastern Spain

Lupinus mariae-josephi is a recently described Lupinus species (Pascual 2004) endemic of a Southeastern area of Spain with soils singularly of high pH and active lime content where it is endangered due to the reduced size of its habitat. Ten isolates of L. mariae-josephi endosymbiotic bacteria were obtained using trap-plants and soils from five sampling points within a native plant population area in Llombai (Valencia, Spain). The microsymbionts are extra-slow (ultrabradytrophic) growing bacteria with phenotypic and symbiotic characteristics singularly different from Bradyrhizobium strains nodulating other Lupinus spp. thriving in the Iberian Peninsula and adapted to growth in acidic soils. Cross-inoculation experiments revealed that these L. mariae-josephi endosymbiotic bacteria isolates are unable to nodulate or efficiently fix nitrogen with other Lupinus spp. Their phylogenetic status was examined by a multilocus sequence analysis of four housekeeping genes (16S rDNA, glnII, recA, atpD) and the symbiotic nodC gene. The 16S rDNA phylogenetic analysis showed that L. mariae-josephi isolates are related to strains nodulating Retama spp. in northeastern Algeria (Boulila et al., 2009), Phaseolus lunatus from Peru (Ormeño-Orrillo et al., 2006), as well as to B. elkanii, B. jicamae and B. pachyrhizi species, forming a new clade (Clade I) within the Bradyrhizobium genus. All the single and concatenated glnII+recA and glnII+recA+atpD analyses consistently support the existence of Clade I, and also revealed that, within this clade, the L. mariae-josephi endosymbiotic bacteria belong to a single evolutionary lineage that also includes strains nodulating Retama spp. from northeastern Algeria. Within this new Bradyrhizobium lineage, the phylogenetic analyses performed showed essentially convergent results indicating that the tested L. mariae-josephi isolates nested in three sub-groups that might correspond to novel sister Bradyrhizobium species. Bradyrhizobium Clade I is highly differentiated from the Bradyrhizobium clade (Clade II) that includes currently named Bradyrhizobium species and well-delineated unnamed genospecies. Singularly, all the endosymbiotic bacteria from Lupinus species adapted to acid soils in the Iberian Peninsula and tested in this study are included in Clade II. They are related either to strains of the B. canariense or B. japonicum lineages. The phylogenetic analysis based on the symbiotic nodC gene showed that L. mariae-josephi endosymbiotic bacteria define a novel branch in the nodC Bradyrhizobium tree. This branch groups together with a branch that gathers isolates from recently studied legume symbiosis such as isolates from Retama spp., which suggests the existence of a common unique ancestor for the symbiotic genes of these two groups of bradyrhizobia. In contrast, the symbiotic genes of isolates from other Lupinus spp. from the Iberian Peninsula are clearly related to the B. canariense lineage. The allopatric (geographic) speciation of the L. mariae-josephi bradyrhizobia may result from the colonization of a singular habitat, such as the basic and high calcium carbonate soils of the Valencia area, by its unique legume host

Phenotypic and molecular diversity of Lupinus mariae-josephi endosymbiotic bacteria.

Lupinus mariae-josephi is a new Lupinus species recently described in a Southeastern area of Spain (Valencia) in soils of singularly high pH and active lime content. Bacteria from L. mariaejosephi have been isolated and correspond to extra-slow growing bacteria symbiotically and phylogenetically distant to endosymbiotic strains nodulating other Lupinus sp. native of the Iberian Peninsula and adapted to growth in acid soils. Cross-inoculation experiments revealed that the L. mariae-josephi endosymbiotic bacteria are unable to nodulate or efficiently fix nitrogen with well-known Lupinus spp. Their species affiliation was examined by a multilocus sequence analysis of four housekeeping genes (16S rDNA, glnII, recA, atpD) and the symbiotic nodC gene. Single and concatenated phylogenetic analyses of these genes consistently revealed that L. mariae-josephi endosymbiotic bacteria belong to a clade, within the Bradyrhizobium genus, highly differentiated from the Bradyrhizobium clade that includes currently named Bradyrhizobium species as well as the endosymbiotic bacteria from Lupinus species tested in this study. Within this new clade the L. mariae-josephi bacteria nested in several subgroups that may correspond to novel sister species. The phylogenetic analysis based on the nodC gene showed that L. mariae-josephi endosymbiotic bacteria define a novel branch in the nodC Bradyrhizobium tree and likely have a common unique ancestor for the symbiotic genes with nodule isolates from Retama spp. At this moment two draft genome sequences belonging to a Bradyrhizobium isolated from L. angustifolius (ISLU101) and to a Bradyrhizobium isolated from L. mariae-josephi (LmjC) have been obtained. The first analysis showed that both genomes correspond to very large chromosomes (>8000 genes) with a high number of unique proteins. nod genes organization are highly conserved among ISLU101, LmjC and B. japonicum USDA110. LmjC presents a single cluster with nod genes from diverse origins. ISLU101 fix genes are found in a single cluster homologous to that of USDA110. LmjC possess a complete copy of fix genes homologous to that from USDA110 and an incomplete one similar to S. meliloti. LmjC possess TypeIII and TypeIV secretion systems. ISLU101 has a Type IV homologous to that of photosynthetic Bradyrhizobium BTAi1, and two copies of Type VI, one homologous to that of USDA110 and the other to R. leguminosarum. Preliminary data indicate that ISLU101 contains a potential second replicon (~100 genes) with high homology to Bradyrhizobium BTai1 plasmid sequenc

Mutational analysis of the RNA-binding domain of the Prunus necrotic ringspot virus (PNRSV) movement protein reveals its requirement for cell-to-cell movement

The movement protein (MP) of Prunus necrotic ringspot virus (PNRSV) is required for cell-to-cell movement. MP subcellular localization studies using a GFP fusion protein revealed highly punctate structures between neighboring cells, believed to represent plasmodesmata. Deletion of the RNA-binding domain (RBD) of PNRSV MP abolishes the cell-to-cell movement. A mutational analysis on this RBD was performed in order to identify in vivo the features that govern viral transport. Loss of positive charges prevented the cell-to-cell movement even though all mutants showed a similar accumulation level in protoplasts to those observed with the wild-type (wt) MP. Synthetic peptides representing the mutants and wild-type RBDs were used to study RNA-binding affinities by EMSA assays being approximately 20-fold lower in the mutants. Circular dichroism analyses revealed that the secondary structure of the peptides was not significantly affected by mutations. The involvement of the affinity changes between the viral RNA and the MP in the viral cell-to-cell movement is discussed

Processing of functional fine scale ceramic structures by ink-jet printing

International audienceThis review illustrates the potentiality of ink-jet printing for the fabrication of functional fine scale ceramic structures corresponding to two different kinds of micro-pillar arrays i.e. (i) PZT skeletons, etc..

Distinct Microglial Responses in Two Transgenic Murine Models of TAU Pathology

Microglial cells are crucial players in the pathological process of neurodegenerative diseases, such as Alzheimer’s disease (AD). Microglial response in AD has been principally studied in relation to amyloid-beta pathology but, comparatively, little is known about inflammatory processes associated to tau pathology. In the hippocampus of AD patients, where tau pathology is more prominent than amyloid-beta pathology, a microglial degenerative process has been reported. In this work, we have directly compared the microglial response in two different transgenic tau mouse models: ThyTau22 and P301S. Surprisingly, these two models showed important differences in the microglial profile and tau pathology. Where ThyTau22 hippocampus manifested mild microglial activation, P301S mice exhibited a strong microglial response in parallel with high phospho-tau accumulation. This differential phospho-tau expression could account for the different microglial response in these two tau strains. However, soluble (S1) fractions from ThyTau22 hippocampus presented relatively high content of soluble phospho-tau (AT8-positive) and were highly toxic for microglial cells in vitro, whereas the correspondent S1 fractions from P301S mice displayed low soluble phospho-tau levels and were not toxic for microglial cells. Therefore, not only the expression levels but the aggregation of phospho-tau should differ between both models. In fact, most of tau forms in the P301S mice were aggregated and, in consequence, forming insoluble tau species. We conclude that different factors as tau mutations, accumulation, phosphorylation, and/or aggregation could account for the distinct microglial responses observed in these two tau models. For this reason, deciphering the molecular nature of toxic tau species for microglial cells might be a promising therapeutic approach in order to restore the deficient immunological protection observed in AD hippocampus