Transgenic potato plants expressing oxalate oxidase have increased resistance to oomycete and bacterial pathogens

Summary: Potato (cv. Bintje) was transformed with a gene encoding an oxalate oxidase from wheat under the control of the CaMV35S promoter. Transgenic potato plants produced high constitutive levels of H2O2 as visualized by 4-chloro-l-naphtol staining. The resistance of these plants was tested againstPhytophthora infestans. An increased level of resistance to the disease was marked by a reduced number of lesions as well as by a decreased number of sporangia formed per lesion. In addition, oxalate oxidase overexpressing plants also exhibited improved resistance toStreptomyces reticuliscabiei, the causal agent of netted scab. Increased expression of oxalate oxidase had no effect on the interaction withErwinia carotovora. These experiments show that overexpression of oxalate oxidase represents a potentially interesting approach for protection of potato to pathogen

A scanner system for high-resolution quantification of variation in root growth dynamics of Brassica rapa genotypes

The potential exists to breed for root system architectures that optimize resource acquisition. However, this requires the ability to screen root system development quantitatively, with high resolution, in as natural an environment as possible, with high throughput. This paper describes the construction of a low-cost, high-resolution root phenotyping platform, requiring no sophisticated equipment and adaptable to most laboratory and glasshouse environments, and its application to quantify environmental and temporal variation in root traits between genotypes of Brassica rapa L. Plants were supplied with a complete nutrient solution through the wick of a germination paper. Images of root systems were acquired without manual intervention, over extended periods, using multiple scanners controlled by customized software. Mixed-effects models were used to describe the sources of variation in root traits contributing to root system architecture estimated from digital images. It was calculated that between one and 43 replicates would be required to detect a significant difference (95% CI 50% difference between traits). Broad-sense heritability was highest for shoot biomass traits (>0.60), intermediate (0.25–0.60) for the length and diameter of primary roots and lateral root branching density on the primary root, and lower (<0.25) for other root traits. Models demonstrate that root traits show temporal variations of various types. The phenotyping platform described here can be used to quantify environmental and temporal variation in traits contributing to root system architecture in B. rapa and can be extended to screen the large populations required for breeding for efficient resource acquisition

Genetic diversity of[i] Rhizoctonia solani[/i] associated with potato tubers in France

Affiche, résuméThe plant pathogenic soil-borne fungus Rhizoctonia solani causes severe damages in crops all around the world. Tubers of potato are frequently affected by R. solani leading to the downgrading of the production. Generally the isolates involved in the sclerotia occurring at the surface of the tuber are assigned to the anastomosis group (AG) 3 but a more precise characterization of the diversity of this deleterious group is needed to set up appropriate control strategies. The diversity of 73 French isolates from the mains potato seed production areas and 31 isolates originating from 9 other countries was assessed according to 3 molecular approaches. Three phylogenetic trees were built up based on the sequences of the internal transcribed spacer (ITS) region and the gene tef-1α as well as the comparison of the total DNA fingerprints of each strain established by amplified fragment length polymorphism (AFLP). The determination of the AGs of R. solani based on the sequencing of the ITS region showed 3 different AGs among our collection (60 AG 3, 8 AG 2-1 and 5 AG 5). Grouping of the isolates belonging to the same AG was confirmed by the sequencing of the gene tef-1α used for the first time to study the genetic diversity of R. solani. About 42 % of the ITS sequences and 73 % of the gene tef-1α sequences contained polymorphic sites where several nucleotides are possible, suggesting that the cells of R. solani strains contain several copies of ITS and gene tef-1α within the same nucleus or between different nuclei. Phylogenetic trees showed a greater genetic diversity within AGs in tef-1α sequences than in ITS sequences. The AFLP analyses showed an even greater diversity among the strains demonstrating that the French strains of R. solani isolated from potatoes were not a clonal population. Moreover, there was no relationship between the geographical origins of the strains or the potato variety from which they were isolated and their genetic diversity. This important and under evaluated genetic diversity as the lack of population structure suggest important genetic mixings leading to a constant evolution within R. solani and could explain the difficulties to control it successfully

Affectation des temps et temporalités dans les couples où l’homme est au foyer – une transgression circonscrite

Les couples où l’homme est au foyer présentent une réponse originale aux contraintes de conciliation travail/famille, par rapport aux normes de genre. Dans cet article, à partir d’une enquête exploratoire auprès de « pères au foyer » menée en France, nous nous intéressons aux temporalités dans lesquelles sont inscrites ces hommes et leurs conjointes. Nous montrerons que contrairement à la majorité des femmes au foyer, le temps de ces pères n’est pas toujours affecté en priorité au foyer, puisque de nombreux pères de notre échantillon investissent leur temps dans des activités rémunérées ou personnelles. Ainsi, les pères disposeraient de la possibilité de marquer leur temps comme étant personnel ou familial (et ce même lorsqu’ils ne font pas autant de tâches ménagères qu’ils estiment devoir le faire), là où leurs conjointes semblent affecter leur temps libre (hors du travail salarié) au foyer et à la famille.Couples with fathers-at-home change the rules of gender, by giving the responsibility of home and children to the man of the family. In this article, from an exploratory investigation about “fathers at home” in France, we will show what uses of time make these fathers and their spouses. Unlike the majority of housewives, the time of the fathers we met is not always allocated as a priority to the home: numerous fathers prefer to do some paid activities or leisure ones. So, fathers can choose if their time is a private one or a family one (even when they don’t do as much housework as they think they should), when their spouses allocate their free time (when they’re not at work) to home and family

Schreiber Michele, American Postfeminist Cinema. Women, Romance and Contemporary Culture

Dans American Postfeminist Cinema, Michele Schreiber questionne les liens entre le genre cinématographique de la romance (les comédies et les drames romantiques) et le postféminisme, en se concentrant sur des films étatsuniens produits entre 1980 et 2012. Cet ouvrage s’inscrit dans la continuité de travaux sur la « rhétorique postféministe » (p. 2), qui « a œuvré à accommoder le féminisme via la représentation de la femme comme consommatrice émancipée ». L’auteure part du constat que dans le ..

The effect of peri-conception hyperglycaemia and the involvement of the hexosamine biosynthesis pathway in mediating oocyte and embryo developmental competence

The environment that the oocyte is exposed to during the peri-conception period can have a significant impact on oocyte developmental competence (the ability of the oocyte to support fertilisation and subsequent embryo development) and the long-term health of the resulting offspring. This is particularly true for maternal hyperglycaemia. While maternal hyperglycaemia during early pregnancy through term development has been extensively studied, the effects on the oocyte itself, and the underlying mechanisms, remain largely unknown. There is increasing evidence, however, for the role of the fuel-sensing hexosamine biosynthesis pathway in mediating the effects of hyperglycaemia in many different cell types. In this review, we will focus on the reproductive consequences of maternal hyperglycaemia during the peri-conceptual period and the role of the hexosamine pathway in mediating these processes.Laura A. Frank, Melanie L. Sutton-McDowall, Robert B. Gilchrist, and Jeremy G. Thompso

Fertilization induces a transient exposure of phosphatidylserine in mouse eggs

Phosphatidylserine (PS) is normally localized to the inner leaflet of the plasma membrane and the requirement of PS translocation to the outer leaflet in cellular processes other than apoptosis has been demonstrated recently. In this work we investigated the occurrence of PS mobilization in mouse eggs, which express flippase Atp8a1 and scramblases Plscr1 and 3, as determined by RT-PCR; these enzyme are responsible for PS distribution in cell membranes. We find a dramatic increase in binding of flouresceinated-Annexin-V, which specifically binds to PS, following fertilization or parthenogenetic activation induced by SrCl2 treatment. This increase was not observed when eggs were first treated with BAPTA-AM, indicating that an increase in intracellular Ca2+ concentration was required for PS exposure. Fluorescence was observed over the entire egg surface with the exception of the regions overlying the meiotic spindle and sperm entry site. PS exposure was also observed in activated eggs obtained from CaMKIIγ null females, which are unable to exit metaphase II arrest despite displaying Ca2+ spikes. In contrast, PS exposure was not observed in TPEN-activated eggs, which exit metaphase II arrest in the absence of Ca2+ release. PS exposure was also observed when eggs were activated with ethanol but not with a Ca2+ ionophore, suggesting that the Ca2+ source and concentration are relevant for PS exposure. Last, treatment with cytochalasin D, which disrupts microfilaments, or jasplakinolide, which stabilizes microfilaments, prior to egg activation showed that PS externalization is an actin-dependent process. Thus, the Ca2+ rise during egg activation results in a transient exposure of PS in fertilized eggs that is not associated with apoptosis.Fil: Curia, Claudio Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Ernesto, Juan Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Stein, Paula. University of Pennsylvania; Estados UnidosFil: Busso, Dolores. Pontificia Universidad Católica de Chile; ChileFil: Schultz, Richard. University of Pennsylvania; Estados UnidosFil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Cohen, Debora Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); Argentin

Assisted hatching in mouse embryos using a noncontact Ho:YSGG laser system

PurposeA noncontact holmium:yttrium scandium gallium garnet (Ho:YSGG) laser system has been designed and tested for the micromanipulation of mammalian embryos. The purpose of this preliminary investigation was to determine the effectiveness of this laser for assisted hatching and evaluate its impact on embryo viability. The Ho:YSGG system, utilizing 250-microsecond pulses at a wavelength of 2.1 microns and 4 Hz, was used to remove a portion of the zona pellucida (ZP) of two- to four-cell FVB mouse embryos.ResultsIn the first experiment there was no difference in blastocyst production or hatching rates following laser or conventional assisted hatching (LAH or AH, respectively) in contrast to control embryos cultured in a 5% CO2 humidified air incubator at 37 degrees C. In the second experiment a blastocyst antihatching culture model was employed and LAH-treated embryos were cultured in a serum-free HTF medium (HTF-o). Blastocyst formation was not influenced by LAH treatment and hatching was increased (P &lt; 0.01) from 4 to 60% compared to HTF-o control group.ConclusionsThese preliminary data demonstrate the utility and nontoxic properties of the Ho:YSGG laser system for quick and precise ZP drilling

Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes

<p>Abstract</p> <p>Background</p> <p>RNA interference (RNAi) is a powerful approach to study a gene function. Transgenic RNAi is an adaptation of this approach where suppression of a specific gene is achieved by expression of an RNA hairpin from a transgene. In somatic cells, where a long double-stranded RNA (dsRNA) longer than 30 base-pairs can induce a sequence-independent interferon response, short hairpin RNA (shRNA) expression is used to induce RNAi. In contrast, transgenic RNAi in the oocyte routinely employs a long RNA hairpin. Transgenic RNAi based on long hairpin RNA, although robust and successful, is restricted to a few cell types, where long double-stranded RNA does not induce sequence-independent responses. Transgenic RNAi in mouse oocytes based on a shRNA offers several potential advantages, including simple cloning of the transgenic vector and an ability to use the same targeting construct in any cell type.</p> <p>Results</p> <p>Here we report our experience with shRNA-based transgenic RNAi in mouse oocytes. Despite optimal starting conditions for this experiment, we experienced several setbacks, which outweigh potential benefits of the shRNA system. First, obtaining an efficient shRNA is potentially a time-consuming and expensive task. Second, we observed that our transgene, which was based on a common commercial vector, was readily silenced in transgenic animals.</p> <p>Conclusions</p> <p>We conclude that, the long RNA hairpin-based RNAi is more reliable and cost-effective and we recommend it as a method-of-choice when a gene is studied selectively in the oocyte.</p

A High-Speed Congenic Strategy Using First-Wave Male Germ Cells

BACKGROUND: In laboratory mice and rats, congenic breeding is essential for analyzing the genes of interest on specific genetic backgrounds and for analyzing quantitative trait loci. However, in theory it takes about 3-4 years to achieve a strain carrying about 99% of the recipient genome at the tenth backcrossing (N10). Even with marker-assisted selection, the so-called 'speed congenic strategy', it takes more than a year at N4 or N5. METHODOLOGY/PRINCIPAL FINDINGS: Here we describe a new high-speed congenic system using round spermatids retrieved from immature males (22-25 days of age). We applied the technique to three genetically modified strains of mice: transgenic (TG), knockin (KI) and N-ethyl-N-nitrosourea (ENU)-induced mutants. The donor mice had mixed genetic backgrounds of C57BL/6 (B6):DBA/2 or B6:129 strains. At each generation, males used for backcrossing were selected based on polymorphic marker analysis and their round spermatids were injected into B6 strain oocytes. Backcrossing was repeated until N4 or N5. For the TG and ENU-mutant strains, the N5 generation was achieved on days 188 and 190 and the proportion of B6-homozygous loci was 100% (74 markers) and 97.7% (172/176 markers), respectively. For the KI strain, N4 was achieved on day 151, all the 86 markers being B6-homozygous as early as on day 106 at N3. The carrier males at the final generation were all fertile and propagated the modified genes. Thus, three congenic strains were established through rapid generation turnover between 41 and 44 days. CONCLUSIONS/SIGNIFICANCE: This new high-speed breeding strategy enables us to produce congenic strains within about half a year. It should provide the fastest protocol for precise definition of the phenotypic effects of genes of interest on desired genetic backgrounds