Strategies for wheat stripe rust pathogenicity identified by transcriptome sequencing

Stripe rust caused by the fungus Puccinia striiformis f.sp. tritici (Pst) is a major constraint to wheat production worldwide. The molecular events that underlie Pst pathogenicity are largely unknown. Like all rusts, Pst creates a specialized cellular structure within host cells called the haustorium to obtain nutrients from wheat, and to secrete pathogenicity factors called effector proteins. We purified Pst haustoria and used next-generation sequencing platforms to assemble the haustorial transcriptome as well as the transcriptome of germinated spores. 12,282 transcripts were assembled from 454-pyrosequencing data and used as reference for digital gene expression analysis to compare the germinated uredinospores and haustoria transcriptomes based on Illumina RNAseq data. More than 400 genes encoding secreted proteins which constitute candidate effectors were identified from the haustorial transcriptome, with two thirds of these up-regulated in this tissue compared to germinated spores. RT-PCR analysis confirmed the expression patterns of 94 effector candidates. The analysis also revealed that spores rely mainly on stored energy reserves for growth and development, while haustoria take up host nutrients for massive energy production for biosynthetic pathways and the ultimate production of spores. Together, these studies substantially increase our knowledge of potential Pst effectors and provide new insights into the pathogenic strategies of this important organism.J.P.R. is an ARC Future Fellow (FT0992129). This project has been supported by Bioplatforms Australia through funding from the Commonwealth Government NCRIS and Education Investment Fund Super Science programs

The ins and outs of rust haustoria

Rust diseases caused by fungi of the order Pucciniales afflict a wide range of plants, including cereals, legumes, ornamentals, and fruit trees, and pose a serious threat to cropping systems and global food security. The obligate parasitic lifestyle of these fungi and their complex life cycles, often involving alternate hosts for the sexual and asexual stages, also make this group of pathogens of great biological interest. One of the most remarkable adaptations of rust fungi is the specialized infection structure that underpins the sustained biotrophic association with hosts; the haustorium.This work has been supported by grants from the TwoBlades corporation and the Australian research council

Comparative genomics of Australian isolates of the wheat stem rust pathogen Puccinia graminis f. sp. tritici reveals extensive polymorphism in candidate effector genes

The wheat stem rust fungus Puccinia graminis f. sp. tritici (Pgt) is one of the most destructive pathogens of wheat. In this study, a draft genome was built for a founder Australian Pgt isolate of pathotype (pt.) 21-0 (collected in 1954) by next generation DNA sequencing. A combination of reference-based assembly using the genome of the previously sequenced American Pgt isolate CDL 75-36-700-3 (p7a) and de novo assembly were performed resulting in a 92 Mbp reference genome for Pgt isolate 21-0. Approximately 13 Mbp of de novo assembled sequence in this genome is not present in the p7a reference assembly. This novel sequence is not specific to 21-0 as it is also present in three other Pgt rust isolates of independent origin. The new reference genome was subsequently used to build a pan-genome based on five Australian Pgt isolates. Transcriptomes from germinated urediniospores and haustoria were separately assembled for pt. 21-0 and comparison of gene expression profiles showed differential expression in ∼10% of the genes each in germinated spores and haustoria. A total of 1,924 secreted proteins were predicted from the 21-0 transcriptome, of which 520 were classified as haustorial secreted proteins (HSPs). Comparison of 21-0 with two presumed clonal field derivatives of this lineage (collected in 1982 and 1984) that had evolved virulence on four additional resistance genes (Sr5, Sr11, Sr27, SrSatu) identified mutations in 25 HSP effector candidates. Some of these mutations could explain their novel virulence phenotypes.Authors wish to thank the Two Blades Foundation for financial support. Part of this work was supported through access to facilities managed by Bioplatforms Australia and funded by the Australian Government National Collaborative Research Infrastructure Strategy and Education Investment Fund Super Science Initiative

“La mejor clase de física de mi vida”: perspectiva del maestro novato

This paper emphasizes the criteria that are at the basis of the argument of the beginning teachers for the Degree in Physics from the Universidad Distrital Francisco Jose de Caldas, to present their experiences in their physical best kind of living. The instrument to identify teachers' ideas is through an open trial, in which they describe their experiences spontaneously. It is intended then by means of an interpretation based on a qualitative and methodological approach to the teaching of physics to characterize some parameters to give a brief generalization affinities majority of students, selecting those most relevantand comparing the findings obtained in some research articulate teacher thinking paradigm and which establish an interpretative approach of the physics teacher discourse, especially on issues such as physical education, the role of teacher, student attitudes and activities of the class physics.En esta ponencia se hace énfasis en los criterios que se encuentran a la base del argumento de los profesores novatos de la Licenciatura en Física de la Universidad Distrital Francisco José de Caldas, para presentar sus experiencias en cuanto a su mejor clase de física vivida. El instrumento para detectar las ideas de los maestros es por medio de una prueba abierta, en la cual ellos describen su experiencia espontáneamente. Se pretende entonces por medio de una interpretación cualitativa y basados en una aproximación metodológica de la enseñanza de la física, caracterizar algunos parámetros para dar una breve generalización a las afinidades mayoritarias de los estudiantes, seleccionando aquellos de mayor relevancia y comparando las conclusiones obtenidas en algunas investigaciones que articulan el paradigma del pensamiento del profesor y que permiten establecer una aproximación interpretativa del discurso del profesor de física, especialmente sobre aspectos como la enseñanza de la física, el papel del profesor, la actitud de los estudiantes y las actividades propias de la clase de física

Discrimination learning with variable stimulus 'salience'.

BACKGROUND: In nature, sensory stimuli are organized in heterogeneous combinations. Salient items from these combinations 'stand-out' from their surroundings and determine what and how we learn. Yet, the relationship between varying stimulus salience and discrimination learning remains unclear. PRESENTATION OF THE HYPOTHESIS: A rigorous formulation of the problem of discrimination learning should account for varying salience effects. We hypothesize that structural variations in the environment where the conditioned stimulus (CS) is embedded will be a significant determinant of learning rate and retention level. TESTING THE HYPOTHESIS: Using numerical simulations, we show how a modified version of the Rescorla-Wagner model, an influential theory of associative learning, predicts relevant interactions between varying salience and discrimination learning. IMPLICATIONS OF THE HYPOTHESIS: If supported by empirical data, our model will help to interpret critical experiments addressing the relations between attention, discrimination and learning

Survey and analysis of microsatellites from transcript sequences in Phytophthora species: frequency, distribution, and potential as markers for the genus

BACKGROUND: Members of the genus Phytophthora are notorious pathogens with world-wide distribution. The most devastating species include P. infestans, P. ramorum and P. sojae. In order to develop molecular methods for routinely characterizing their populations and to gain a better insight into the organization and evolution of their genomes, we used an in silico approach to survey and compare simple sequence repeats (SSRs) in transcript sequences from these three species. We compared the occurrence, relative abundance, relative density and cross-species transferability of the SSRs in these oomycetes. RESULTS: The number of SSRs in oomycetes transcribed sequences is low and long SSRs are rare. The in silico transferability of SSRs among the Phytophthora species was analyzed for all sets generated, and primers were selected on the basis of similarity as possible candidates for transferability to other Phytophthora species. Sequences encoding putative pathogenicity factors from all three Phytophthora species were also surveyed for presence of SSRs. However, no correlation between gene function and SSR abundance was observed. The SSR survey results, and the primer pairs designed for all SSRs from the three species, were deposited in a public database. CONCLUSION: In all cases the most common SSRs were trinucleotide repeat units with low repeat numbers. A proportion (7.5%) of primers could be transferred with 90% similarity between at least two species of Phytophthora. This information represents a valuable source of molecular markers for use in population genetics, genetic mapping and strain fingerprinting studies of oomycetes, and illustrates how genomic databases can be exploited to generate data-mining filters for SSRs before experimental validation

Re-programming mouse liver-resident invariant natural killer T cells for suppressing hepatic and diabetogenic autoimmunity

Invariant NKT (iNKT) cells comprise a heterogeneous group of non-circulating, tissue-resident T lymphocytes that recognize glycolipids, including alpha-galactosylceramide (?GalCer), in the context of CD1d, but whether peripheral iNKT cell subsets are terminally differentiated remains unclear. Here we show that mouse and human liver-resident ?GalCer/CD1d-binding iNKTs largely correspond to a novel Zbtb16+Tbx21+Gata3+MaflowRorc- subset that exhibits profound transcriptional, phenotypic and functional plasticity. Repetitive in vivo encounters of these liver iNKT (LiNKT) cells with intravenously delivered ?GalCer/CD1d-coated nanoparticles (NP) trigger their differentiation into immunoregulatory, IL-10+IL-21-producing Zbtb16highMafhighTbx21+Gata3+Rorc- cells, termed LiNKTR1, expressing a T regulatory type 1 (TR1)-like transcriptional signature. This response is LiNKT-specific, since neither lung nor splenic tissue-resident iNKT cells from ?GalCer/CD1d-NP-treated mice produce IL-10 or IL-21. Additionally, these LiNKTR1 cells suppress autoantigen presentation, and recognize CD1d expressed on conventional B cells to induce IL-10+IL-35-producing regulatory B (Breg) cells, leading to the suppression of liver and pancreas autoimmunity. Our results thus suggest that LiNKT cells are plastic for further functional diversification, with such plasticity potentially targetable for suppressing tissue-specific inflammatory phenomena.© 2022. The Author(s)

Evanescent-Mode Ridge-Waveguide Radiating Filters for Space Applications

[EN] This paper describes a new family of all-metallic radiators based on below-cutoff apertures fed by evanescentmode ridge waveguide filters. The pass band of the filters and the polarization of the apertures are configured to radiate orthogonal signals at different center frequencies. The simultaneous operation of these radiators in an array configuration results in interleaved apertures producing overlapped fields. This strategy reduces the volume and weight of the antenna system, which is a specially appealing characteristic in satellite antennas. Each isolated evanescent-mode filter ends in a small-size aperture and is designed following a novel synthesis procedure. Next, the radiating elements are combined, and the resulting array is analyzed as a multiplexer from the input ports to the freespace region thus accounting for all mutual couplings between different apertures. To validate the theoretical formulations, a prototype has also been manufactured. The results obtained show very good agreement between simulation and measurements in terms of reflection coefficient, side-lobe level and realized gain, thereby fully validating the new family of radiating arrays.This work was supported by the Spanish Ministry of Economics and Competitiveness under Project TEC2016-79700-C2-1-R, Project TEC2016-78028-C3-3-P, Project TEC2013-47037-C5-1-R, and Project TEC2016-75934-C4-1-R.Sánchez-Escuderos, D.; Ruiz-Garnica, J.; Baquero Escudero, M.; Soto Pacheco, P.; Boria Esbert, VE.; Tosso, G.; Angeletti, P.... (2019). Evanescent-Mode Ridge-Waveguide Radiating Filters for Space Applications. IEEE Transactions on Antennas and Propagation. 67(10):6286-6297. https://doi.org/10.1109/TAP.2019.2920272S62866297671

Study of Microscopic Residual Stresses in an Extruded Aluminium Alloy Sample after Thermal Treatment

Abstract: A method is proposed to calculate the microscopic residual stresses in extruded cylindrical samples of non-ageing aluminium alloy 5083 (Al–Mg), arising from quenching in fresh water from 530°C. We start from the premise that the alloy is single-phase and non-isotropic on a microscopic scale; it consists of many grains that exhibit different mechanical response depending on their crystallographic orientation and neighboring grains. Microscopic residual stresses depend on the applied heat treatment, microstructure and mechanical strength of the individual grains. The stresses were calculated from neutron diffraction data. Genetic programming algorithms were used to calculate microscopic residual stresses, considering that each diffraction peak describes the stress distribution of a group of grains having a certain orientation, size and environment. The algorithm assigns a stress value to each grain according to the distribution of the diffraction peaks and the microstructural parameters of these grains.This work was supported by the Madrid Regional Government-FEDER grant Y2018/NMT-4668 (Micro-Stress-MAP-CM) and the project MAT2017-83825-C4-1-R. We would also like to express our gratitude to FLNR-JINP for the beam time allocated on the FSD instrument, and to the HeuristicLab Software developers