A Mediator subunit imparts robustness to a polyphenism decision.

Polyphenism is a type of developmental plasticity that translates continuous environmental variability into discontinuous phenotypes. Such discontinuity likely requires a switch between alternative gene-regulatory networks, a principle that has been borne out by mechanisms found to promote morph-specific gene expression. However, whether robustness is required to execute a polyphenism decision has awaited testing at the molecular level. Here, we used a nematode model for polyphenism, Pristionchus pacificus, to identify the molecular regulatory factors that ensure the development of alternative forms. This species has a dimorphism in its adult feeding structures, specifically teeth, which are a morphological novelty that allows predation on other nematodes. Through a forward genetic screen, we determined that a duplicate homolog of the Mediator subunit MDT-15/MED15, P. pacificus MDT-15.1, is necessary for the polyphenism and the robustness of the resulting phenotypes. This transcriptional coregulator, which has a conserved role in metabolic responses to nutritional stress, coordinates these processes with its effects on this diet-induced polyphenism. Moreover, this MED15 homolog genetically interacts with two nuclear receptors, NHR-1 and NHR-40, to achieve dimorphism: Single and double mutants for these three factors result in morphologies that together produce a continuum of forms between the extremes of the polyphenism. In summary, we have identified a molecular regulator that confers discontinuity to a morphological polyphenism, while also identifying a role for MED15 as a plasticity effector.

Toxicological impacts of microplastics on virulence, reproduction and physiological process of entomopathogenic nematodes.

Microplastics have emerged as significant and concerning pollutants within soil ecosystems. Among the soil biota, entomopathogenic nematodes (EPNs) are lethal parasites of arthropods, and are considered among the most effective biological agents against pests. Infective juveniles (IJs) of EPNs, as they navigate the soil matrix scavenging for arthropod hosts to infect, they could potentially encounter microplastics. Howver, the impact of microplastics on EPNs has not been fully elucidated yet. We addressed this gap by subjecting Steinernema feltiae EPNs to polystyrene microplastics (PS-MPs) with various sizes, concentrations, and exposure durations. After confirming PS-MP ingestion by S. feltiae using fluorescent dyes, we found that the PS-MPs reduced the survival, reproduction, and pathogenicity of the tested EPNs, with effects intensifying for smaller PS-MPs (0.1-1 µm) at higher concentrations (105 µg/L). Furthermore, exposure to PS-MPs triggered oxidative stress in S. feltiae, leading to increased reactive oxygen species levels, compromised mitochondrial membrane potential, and increased antioxidative enzyme activity. Furthermore, transcriptome analyses revealed PS-MP-induced suppression of mitochondrial function and oxidative phosphorylation pathways. In conclusion, we show that ingestion of PS-MPs by EPNs can compromise their fitness, due to multple toxicity effects. Our results bear far-reaching consequences, as the presence of microplastics in soil ecosystems could undermine the ecological role of EPNs in regulating pest populations.

Conserved nuclear hormone receptors controlling a novel plastic trait target fast-evolving genes expressed in a single cell.

Environment shapes development through a phenomenon called developmental plasticity. Deciphering its genetic basis has potential to shed light on the origin of novel traits and adaptation to environmental change. However, molecular studies are scarce, and little is known about molecular mechanisms associated with plasticity. We investigated the gene regulatory network controlling predatory vs. non-predatory dimorphism in the nematode Pristionchus pacificus and found that it consists of genes of extremely different age classes. We isolated mutants in the conserved nuclear hormone receptor nhr-1 with previously unseen phenotypic effects. They disrupt mouth-form determination and result in animals combining features of both wild-type morphs. In contrast, mutants in another conserved nuclear hormone receptor nhr-40 display altered morph ratios, but no intermediate morphology. Despite divergent modes of control, NHR-1 and NHR-40 share transcriptional targets, which encode extracellular proteins that have no orthologs in Caenorhabditis elegans and result from lineage-specific expansions. An array of transcriptional reporters revealed co-expression of all tested targets in the same pharyngeal gland cell. Major morphological changes in this gland cell accompanied the evolution of teeth and predation, linking rapid gene turnover with morphological innovations. Thus, the origin of feeding plasticity involved novelty at the level of genes, cells and behavior.

Study on host-seeking behavior and chemotaxis of entomopathogenic nematodes using Pluronic F-127 gel.

Pluronic F-127 gel (PF127) has proven to be a powerful medium in which to study host-finding behavior and chemotaxis for plant-parasitic nematodes. Pluronic gel can also be used to study host-habitat seeking behavior of entomopathogenic nematodes (EPN), which are natural enemies of root-feeding insect pests. In this study, PF127 was used to study tritrophic interactions among EPNs, host-habitat roots and insects. We also tested whether EPN aggregated to acetic acid (pH gradient) which mimicked the conditions near the roots. The chive root gnat Bradysia odoriphaga alone significantly attracted more nematodes than chive roots alone or the combination of roots plus insects. The attractiveness of B. odoriphaga differed (3.7-15.4%) among all tested species/strains of EPNs. In addition, we found that Heterorhabditis spp. and Steinernema spp. infective juveniles responded to pH gradients formed by acetic acid in Pluronic gel. The preferred pH ranges for strains of H. bacteriophora and H. megidis were from 4.32-5.04, and from 5.37-6.92 for Steinernema species, indicating that Heterorhabditis spp. prefer low pH conditions than Steinernema species. A narrow pH gradient between 6.84 and 7.05 was detected around chive root tips in which EPN was attracted. These results suggest that Pluronic gel can be broadly used for the study of host or host-habitat seeking behaviors and chemotaxis of nematodes.

Adaptive value of a predatory mouth-form in a dimorphic nematode.

Polyphenisms can be adaptations to environments that are heterogeneous in space and time, but to persist they require conditional-specific advantages. The nematode Pristionchus pacificus is a facultative predator that displays an evolutionarily conserved polyphenism of its mouthparts. During development, P. pacificus irreversibly executes either a eurystomatous (Eu) or stenostomatous (St) mouth-form, which differ in the shape and number of movable teeth. The Eu form, which has an additional tooth, is more complex than the St form and is thus more highly derived relative to species lacking teeth. Here, we investigate a putative fitness trade-off for the alternative feeding-structures of P. pacificus. We show that the complex Eu form confers a greater ability to kill prey. When adults were provided with a prey diet, Eu nematodes exhibited greater fitness than St nematodes by several measures, including longevity, offspring survival and fecundity when followed by bacterial feeding. However, the two mouth-forms had similar fecundity when fed ad libitum on bacteria, a condition that would confer benefit on the more rapidly developing St form. Thus, the two forms show conditional fitness advantages in different environments. This study provides, to our knowledge, the first functional context for dimorphism in a model for the genetics of plasticity.

Tarantobelus arachnicida (Nematoda: Rhabditida) invasion of exotic spiders in Poland

Nematodes of the Brevibuccidae family were stated among spiders of the Theraphosidae family (the South American species), which were bred in Poland. The first sign of infection was anorexia which led to gradually increasing lethargy progressed to a huddled posture. Additionally, a white discharge with nematodes between mouth and chelicerae was noted. All of the derived nematodes were morphologically identified and determined to the species Tarantobelus arachnicida. A molecular analysis covered amplification and sequencing of small subunit ribosomal RNA (18S rRNA). A post mortem examination demonstrated the presence of nematodes not only near the chelicerae, but also inside the intestine, hence the source of infection might be insects used as a food. The research showed that such kind of infection is an important disease, which poses a serious risk to the breeding spiders. To date there is no effective treatment, however, we demonstrated that usage of the Lugol's solution seems to be promising.

Serotonin Drives Predatory Feeding Behavior via Synchronous Feeding Rhythms in the Nematode Pristionchus pacificus.

Feeding behaviors in a wide range of animals are regulated by the neurotransmitter serotonin, although the exact neural circuits and associated mechanism are often unknown. The nematode Pristionchus pacificus can kill other nematodes by opening prey cuticles with movable teeth. Previous studies showed that exogenous serotonin treatment induces a predatory-like tooth movement and slower pharyngeal pumping in the absence of prey; however, physiological functions of serotonin during predation and other behaviors in P. pacificus remained completely unknown. Here, we investigate the roles of serotonin by generating mutations in Ppa-tph-1 and Ppa-bas-1, two key serotonin biosynthesis enzymes, and by genetic ablation of pharynx-associated serotonergic neurons. Mutations in Ppa-tph-1 reduced the pharyngeal pumping rate during bacterial feeding compared with wild-type. Moreover, the loss of serotonin or a subset of serotonergic neurons decreased the success of predation, but did not abolish the predatory feeding behavior completely. Detailed analysis using a high-speed camera revealed that the elimination of serotonin or the serotonergic neurons disrupted the timing and coordination of predatory tooth movement and pharyngeal pumping. This loss of synchrony significantly reduced the efficiency of successful predation events. These results suggest that serotonin has a conserved role in bacterial feeding and in addition drives the feeding rhythm of predatory behavior in Pristionchus.

Efficacy of Entomopathogenic Nematodes and Sprayable Polymer Gel Against Crucifer Flea Beetle (Coleoptera: Chrysomelidae) on Canola.

The crucifer flea beetle, Phyllotreta cruciferae (Goeze), is a key pest of canola (Brassica napus L.) in the northern Great Plains of North America. The efficacies of entomopathogenic nematodes (Steinernema spp. and Heterorhabditis spp.), a sprayable polymer gel, and a combination of both were assessed on canola for flea beetle management. Plots were treated soon after colonization by adult flea beetles, when canola was in the cotyledon to one-leaf stage. Ten plants along a 3.6-m section of row were selected and rated at pre-treatment and 7 and 14 d post treatment using the damage-rating scheme advanced by the European Plant Protection Organization, where 1 = 0%, 2 = 2%, 3 = 5%, 4 = 10%, and 5 = 25% leaf area injury. Under moderate flea beetle feeding pressure (1-3.3% leaf area damaged), seeds treated with Gaucho 600 (Bayer CropScience LP Raleigh, NC) (imidacloprid) produced the highest yield (843.2 kg/ha). Meanwhile, Barricade (Barricade International, Inc. Hobe Sound, FL) (polymer gel; 1%) + Scanmask (BioLogic Company Inc, Willow Hill, PA) (Steinernema feltiae) resulted in the highest yields: 1020.8 kg/ha under high (2.0-5.3% leaf area damaged), and 670.2 kg/ha at extremely high (4.3-8.6 % leaf area damaged) feeding pressure. Our results suggest that Barricade (1%) + Scanmask (S. feltiae) can serve as an alternative to the conventional chemical seed treatment. Moreover, Scanmask (S. feltiae) can be used to complement the effects of seed treatment after its protection has run out.