Molecular diversity of the Metarhizium anisopliae lineage in an agricultural field

Entomopathogenic fungal isolates identified by morphology as Metarhizium anisopliae may belong to different species when identified by molecular characters. We isolated Metarhizium spp. from an experimental agricultural field under both conventional and organic farming regimes using Tenebrio molitor as bait insect to assess the molecular diversity within the soil. Isolates were analyzed using DNA sequencing and applying SSR markers. Within the former M. anisopliae lineage, we found M. brunneum (86.3%), M. robertsii (11.3%) and M. majus (3.4%) in the soil samples. Several genotypes of each species were identified based on SSR markers. Differences in abundance of the species and their genotypes suggest different adaptations to the soil environment of the agricultural field. There were no effects of conventinal or organic farming regimes on diversity of the fungi

A Robust and Sensitive Synthetic Sensor to Monitor the Transcriptional Output of the Cytokinin Signaling Network in Planta

Cytokinins are classic plant hormones that orchestrate plant growth, development, and physiology. They affect gene expression in target cells by activating a multistep phosphorelay network. Type-B response regulators, acting as transcriptional activators, mediate the final step in the signaling cascade. Previously, we have introduced a synthetic reporter, Two Component signaling Sensor (TCS)::green fluorescent protein (GFP), which reflects the transcriptional activity of type-B response regulators. TCS::GFP was instrumental in uncovering roles of cytokinin and deepening our understanding of existing functions. However, TCS-mediated expression of reporters is weak in some developmental contexts where cytokinin signaling has a documented role, such as in the shoot apical meristem or in the vasculature of Arabidopsis (Arabidopsis thaliana). We also observed that GFP expression becomes rapidly silenced in TCS::GFP transgenic plants. Here, we present an improved version of the reporter, TCS new (TCSn), which, compared with TCS, is more sensitive to phosphorelay signaling in Arabidopsis and maize (Zea mays) cellular assays while retaining its specificity. Transgenic Arabidopsis TCSn::GFP plants exhibit strong and dynamic GFP expression patterns consistent with known cytokinin functions. In addition, GFP expression has been stable over generations, allowing for crosses with different genetic backgrounds. Thus, TCSn represents a significant improvement to report the transcriptional output profile of phosphorelay signaling networks in Arabidopsis, maize, and likely other plants that display common response regulator DNA-binding specificities

Virulence of in vivo and in vitro produced conidia of Metarhizium brunneum strains for control of wireworms

Wireworms are the soil inhabiting larvae of click beetles and can cause severe damage to arable crops such as potatoes (Solanum tuberosum, L.). Several strains of the entomopathogenic fungus Metarhizium brunneum (Petch) are pathogenic to wireworms. In this study, three European strains of M. brunneum were tested in the laboratory against the most damaging wireworm species in Europe, Agriotes lineatus (L.), Agriotes obscurus (L.) and Agriotes sputator (L.). A Swiss strain, isolated from an A. obscurus cadaver, proved to be most effective, killing up to 73% of A. lineatus and 83% A. obscurus individuals, respectively. The median lethal time (LT50) was 21 days post inoculation (dpi) for A. lineatus and 14 dpi for A. obscurus. The strain did not lose virulence through subsequent cultivation on artificial medium and thus seems to be suitable for mass production as a biocontrol agent for wireworm control

Assessing potential hybridization between a hypothetical gene drive-modified Drosophila suzukii and nontarget Drosophila species.

Genetically engineered gene drives (geGD) are potentially powerful tools for suppressing or even eradicating populations of pest insects. Before living geGD insects can be released into the environment, they must pass an environmental risk assessment to ensure that their release will not cause unacceptable harm to non-targeted entities of the environment. A key research question concerns the likelihood that nontarget species will acquire the functional GD elements; such acquisition could lead to reduced abundance or loss of those species and to a disruption of the ecosystem services they provide. The main route for gene flow is through hybridization between the geGD insect strain and closely related species that co-occur in the area of release and its expected dispersal. Using the invasive spotted-wing drosophila, Drosophila suzukii, as a case study, we provide a generally applicable strategy on how a combination of interspecific hybridization experiments, behavioral observations, and molecular genetic analyses can be used to assess the potential for hybridization

Temporal metabolic profiling of theQuercus suber-Phytophthora cinnamomisystem by middle-infrared spectroscopy

The oomycete Phytophthora cinnamomi is an aggressive plant pathogen, detrimental to many ecosystems including cork oak (Quercus suber) stands, and can inflict great losses in one of the greatest ‘hotspots’ for biodiversity in the world. Here, we applied Fourier transform-infrared (FT-IR) spectroscopy combined with chemometrics to disclose the metabolic patterns of cork oak roots and P. cinnamomi mycelium during the early hours of the interaction. As early as 2 h post-inoculation (hpi), cork oak roots showed altered metabolic patterns with significant variations for regions associated with carbohydrate, glycoconjugate and lipid groups when compared to mockinoculated plants. These variations were further extended at 8 hpi. Surprisingly, at 16 hpi, the metabolic changes in inoculated and mock-inoculated plants were similar, and at 24 hpi, the metabolic patterns of the regions mentioned above were inverted when compared to samples collected at 8 hpi. Principal component analysis of the FT-IR spectra confirmed that the metabolic patterns of inoculated cork oak roots could be readily distinguished from those of mock-inoculated plants at 2, 8 and 24 hpi, but not at 16 hpi. FT-IR spectral analysis from mycelium of P. cinnamomi exposed to cork oak root exudates revealed contrasting variations for regions associated with protein groups at 16 and 24 h post-exposure (hpe), whereas carbohydrate and glycoconjugate groups varied mainly at 24 hpe. Our results revealed early alterations in the metabolic patterns of the host plant when interacting with the biotrophic pathogen. In addition, the FTIR technique can be successfully applied to discriminate infected cork oak plants from mock-inoculated plants, although these differences were dynamic with time. To a lesser extent, the metabolic patterns of P. cinnamomi were also altered when exposed to cork oak root exudates

Entwicklung eines generischen Gemeindereferenzprozess-modells in Anlehnung an das ARIS-Modell

Die durchgängige Einführung von E-Government geht im Gegensatz zum Thema E- Business in der Privatwirtschaft eher beschaulich voran. Dies hängt u.a. auch damit zusammen, dass an der Schnittstelle von Verwaltung und IT noch größerer Entwicklungsbedarf ersichtlich ist. Konzeptionell ist dazu noch einige Grundlagenarbeit zu leisten, bis hier mehr Konsistenz in eine Verwaltungseinheits-übergreifende Entwicklung entsteht, sodass sich Interoperabilität zu einem durchgängig umgesetzten Prinzip etabliert. Dieser Beitrag schildert ein integriertes Konzept aus Sicht von Gemeindeverwaltungen, das aber auf allen föderalen Ebenen umgesetzt werden kann. Das Konzept dient der Abbildung von Geschäftsprozessen und den damit verbundenen Aspekten wie Formulare, Informationen, Leistungen, Aufgaben-, Kompetenzund Veranwortungs-Zuordnungen sowie Organisationsund Governance-Aspekten. Als Grundlage dazu dient das für das vorliegende Vorhaben adaptierte ARIS-Modell. In einem iterativen Vorgehen ist das Modell mit Gemeindemitarbeitenden auf die Bedürfnisse einer Gemeinde angepasst worden. Die vorliegende Prozesslandkarte ist in die Bereiche Governance-, Führungs-, Kernund Unterstützungsaufgaben gegliedert. Differenziert werden die folgenden Sichten auf Prozessebene: Eigentliche Prozesssicht, Organisationssicht mit Aufgaben, Verantwortungen und Zuständigkeiten, Informationssicht mit Formularen, Dokumenten und relevanten Gesetzen, Sicht auf Kontrollflüsse, Gateways und Geschäftsregeln sowie die Sicht auf die vom Prozessablauf betroffenen IT-Systeme

Insect pathogens as biological control agents: back to the future

The development and use of entomopathogens as classical, conservation and augmentative biological control agents have included a number of successes and some setbacks in the past 15 years. In this forum paper we present current information on development, use and future directions of insect-specific viruses, bacteria, fungi and nematodes as components of integrated pest management strategies for control of arthropod pests of crops, forests, urban habitats, and insects of medical and veterinary importance. Insect pathogenic viruses are a fruitful source of MCAs, particularly for the control of lepidopteran pests. Most research is focused on the baculoviruses, important pathogens of some globally important pests for which control has become difficult due to either pesticide resistance or pressure to reduce pesticide residues. Baculoviruses are accepted as safe, readily mass produced, highly pathogenic and easily formulated and applied control agents. New baculovirus products are appearing in many countries and gaining an increased market share. However, the absence of a practical in vitro mass production system, generally higher production costs, limited post application persistence, slow rate of kill and high host specificity currently contribute to restricted use in pest control. Overcoming these limitations are key research areas for which progress could open up use of insect viruses to much larger markets. A small number of entomopathogenic bacteria have been commercially developed for control of insect pests. These include several Bacillus thuringiensis sub-species, Lysinibacillus (Bacillus) sphaericus, Paenibacillus spp. and Serratia entomophila. B. thuringiensis sub-species kurstaki is the most widely used for control of pest insects of crops and forests, and B. thuringiensis sub-species israelensis and L. sphaericus are the primary pathogens used for medically important pests including dipteran vectors,. These pathogens combine the advantages of chemical pesticides and microbial control agents (MCAs): they are fast acting, easy to produce at a relatively low cost, easy to formulate, have a long shelf life and allow delivery using conventional application equipment and systemics (i.e. in transgenic plants). Unlike broad spectrum chemical pesticides, B. thuringiensis toxins are selective and negative environmental impact is very limited. Of the several commercially produced MCAs, B. thuringiensis (Bt) has more than 50% of market share. Extensive research, particularly on the molecular mode of action of Bt toxins, has been conducted over the past two decades. The Bt genes used in insect-resistant transgenic crops belong to the Cry and vegetative insecticidal protein families of toxins. Bt has been highly efficacious in pest management of corn and cotton, drastically reducing the amount of broad spectrum chemical insecticides used while being safe for consumers and non-target organisms. Despite successes, the adoption of Bt crops has not been without controversy. Although there is a lack of scientific evidence regarding their detrimental effects, this controversy has created the widespread perception in some quarters that Bt crops are dangerous for the environment. In addition to discovery of more efficacious isolates and toxins, an increase in the use of Bt products and transgenes will rely on innovations in formulation, better delivery systems and ultimately, wider public acceptance of transgenic plants expressing insect-specific Bt toxins. Fungi are ubiquitous natural entomopathogens that often cause epizootics in host insects and possess many desirable traits that favor their development as MCAs. Presently, commercialized microbial pesticides based on entomopathogenic fungi largely occupy niche markets. A variety of molecular tools and technologies have recently allowed reclassification of numerous species based on phylogeny, as well as matching anamorphs (asexual forms) and teleomorphs (sexual forms) of several entomopathogenic taxa in the Phylum Ascomycota. Although these fungi have been traditionally regarded exclusively as pathogens of arthropods, recent studies have demonstrated that they occupy a great diversity of ecological niches. Entomopathogenic fungi are now known to be plant endophytes, plant disease antagonists, rhizosphere colonizers, and plant growth promoters. These newly understood attributes provide possibilities to use fungi in multiple roles. In addition to arthropod pest control, some fungal species could simultaneously suppress plant pathogens and plant parasitic nematodes as well as promote plant growth. A greater understanding of fungal ecology is needed to define their roles in nature and evaluate their limitations in biological control. More efficient mass production, formulation and delivery systems must be devised to supply an ever increasing market. More testing under field conditions is required to identify effects of biotic and abiotic factors on efficacy and persistence. Lastly, greater attention must be paid to their use within integrated pest management programs; in particular, strategies that incorporate fungi in combination with arthropod predators and parasitoids need to be defined to ensure compatibility and maximize efficacy. Entomopathogenic nematodes (EPNs) in the genera Steinernema and Heterorhabditis are potent MCAs. Substantial progress in research and application of EPNs has been made in the past decade. The number of target pests shown to be susceptible to EPNs has continued to increase. Advancements in this regard primarily have been made in soil habitats where EPNs are shielded from environmental extremes, but progress has also been made in use of nematodes in above-ground habitats owing to the development of improved protective formulations. Progress has also resulted from advancements in nematode production technology using both in vivo and in vitro systems; novel application methods such as distribution of infected host cadavers; and nematode strain improvement via enhancement and stabilization of beneficial traits. Innovative research has also yielded insights into the fundamentals of EPN biology including major advances in genomics, nematode-bacterial symbiont interactions, ecological relationships, and foraging behavior. Additional research is needed to leverage these basic findings toward direct improvements in microbial control