A male-killing Wolbachia endosymbiont is concealed by another endosymbiont and a nuclear suppressor.

Bacteria that live inside the cells of insect hosts (endosymbionts) can alter the reproduction of their hosts, including the killing of male offspring (male killing, MK). MK has only been described in a few insects, but this may reflect challenges in detecting MK rather than its rarity. Here, we identify MK Wolbachia at a low frequency (around 4%) in natural populations of Drosophila pseudotakahashii. MK Wolbachia had a stable density and maternal transmission during laboratory culture, but the MK phenotype which manifested mainly at the larval stage was lost rapidly. MK Wolbachia occurred alongside a second Wolbachia strain expressing a different reproductive manipulation, cytoplasmic incompatibility (CI). A genomic analysis highlighted Wolbachia regions diverged between the 2 strains involving 17 genes, and homologs of the wmk and cif genes implicated in MK and CI were identified in the Wolbachia assembly. Doubly infected males induced CI with uninfected females but not females singly infected with CI-causing Wolbachia. A rapidly spreading dominant nuclear suppressor genetic element affecting MK was identified through backcrossing and subsequent analysis with ddRAD SNPs of the D. pseudotakahashii genome. These findings highlight the complexity of nuclear and microbial components affecting MK endosymbiont detection and dynamics in populations and the challenges of making connections between endosymbionts and the host phenotypes affected by them.

Practical Guide to Measuring Wetland Carbon Pools and Fluxes.

Wetlands cover a small portion of the world, but have disproportionate influence on global carbon (C) sequestration, carbon dioxide and methane emissions, and aquatic C fluxes. However, the underlying biogeochemical processes that affect wetland C pools and fluxes are complex and dynamic, making measurements of wetland C challenging. Over decades of research, many observational, experimental, and analytical approaches have been developed to understand and quantify pools and fluxes of wetland C. Sampling approaches range in their representation of wetland C from short to long timeframes and local to landscape spatial scales. This review summarizes common and cutting-edge methodological approaches for quantifying wetland C pools and fluxes. We first define each of the major C pools and fluxes and provide rationale for their importance to wetland C dynamics. For each approach, we clarify what component of wetland C is measured and its spatial and temporal representativeness and constraints. We describe practical considerations for each approach, such as where and when an approach is typically used, who can conduct the measurements (expertise, training requirements), and how approaches are conducted, including considerations on equipment complexity and costs. Finally, we review key covariates and ancillary measurements that enhance the interpretation of findings and facilitate model development. The protocols that we describe to measure soil, water, vegetation, and gases are also relevant for related disciplines such as ecology. Improved quality and consistency of data collection and reporting across studies will help reduce global uncertainties and develop management strategies to use wetlands as nature-based climate solutions. Supplementary Information: The online version contains supplementary material available at 10.1007/s13157-023-01722-2.

High duty cycle moth sounds jam bat echolocation: bats counter with compensatory changes in buzz duration.

Tiger moth species vary greatly in the number of clicks they produce and the resultant duty cycle. Signals with higher duty cycles are expected to more effectively interfere with bat sonar. However, little is known about the minimum duty cycle of tiger moth signals for sonar jamming. Is there a threshold that allows us to classify moths as acoustically aposematic versus sonar jammers based on their duty cycles? We performed playback experiments with three wild-caught adult male bats, Eptesicus fuscus. Bat attacks on tethered moths were challenged using acoustic signals of Bertholdia trigona with modified duty cycles ranging from 0 to 46%. We did not find evidence for a duty cycle threshold; rather, the ability to jam the bat's sonar was a continuous function of duty cycle consistent with a steady increase in the number of clicks arriving during a critical signal processing time window just prior to the arrival of an echo. The proportion of successful captures significantly decreased as the moth duty cycle increased. Our findings suggest that moths cannot be unambiguously classified as acoustically aposematic or sonar jammers based solely on duty cycle. Bats appear to compensate for sonar jamming by lengthening the duration of their terminal buzz and they are more successful in capturing moths when they do so. In contrast to previous findings for bats performing difficult spatial tasks, the number of sonar sound groups decreased in response to high duty cycles and did not affect capture success.

A phylogeny for the Drosophila montium species group: A model clade for comparative analyses.

The Drosophila montium species group is a clade of 94 named species, closely related to the model species D. melanogaster. The montium species group is distributed over a broad geographic range throughout Asia, Africa, and Australasia. Species of this group possess a wide range of morphologies, mating behaviors, and endosymbiont associations, making this clade useful for comparative analyses. We use genomic data from 42 available species to estimate the phylogeny and relative divergence times within the montium species group, and its relative divergence time from D. melanogaster. To assess the robustness of our phylogenetic inferences, we use 3 non-overlapping sets of 20 single-copy coding sequences and analyze all 60 genes with both Bayesian and maximum likelihood methods. Our analyses support monophyly of the group. Apart from the uncertain placement of a single species, D. baimaii, our analyses also support the monophyly of all seven subgroups proposed within the montium group. Our phylograms and relative chronograms provide a highly resolved species tree, with discordance restricted to estimates of relatively short branches deep in the tree. In contrast, age estimates for the montium crown group, relative to its divergence from D. melanogaster, depend critically on prior assumptions concerning variation in rates of molecular evolution across branches, and hence have not been reliably determined. We discuss methodological issues that limit phylogenetic resolution - even when complete genome sequences are available - as well as the utility of the current phylogeny for understanding the evolutionary and biogeographic history of this clade.

Spontaneous motor-behavior abnormalities in two Drosophila models of neurodevelopmental disorders.

Mutations in hundreds of genes cause neurodevelopmental disorders with abnormal motor behavior alongside cognitive deficits. Boys with fragile X syndrome (FXS), a leading monogenic cause of intellectual disability, often display repetitive behaviors, a core feature of autism. By direct observation and manual analysis, we characterized spontaneous-motor-behavior phenotypes of Drosophila dfmr1 mutants, an established model for FXS. We recorded individual 1-day-old adult flies, with mature nervous systems and prior to the onset of aging, in small arenas. We scored behavior using open-source video-annotation software to generate continuous activity timelines, which were represented graphically and quantitatively. Young dfmr1 mutants spent excessive time grooming, with increased bout number and duration; both were rescued by transgenic wild-type dfmr1+. By two grooming-pattern measures, dfmr1-mutant flies showed elevated repetitions consistent with perseveration, which is common in FXS. In addition, the mutant flies display a preference for grooming posterior body structures, and an increased rate of grooming transitions from one site to another. We raise the possibility that courtship and circadian rhythm defects, previously reported for dfmr1 mutants, are complicated by excessive grooming. We also observed significantly increased grooming in CASK mutants, despite their dramatically decreased walking phenotype. The mutant flies, a model for human CASK-related neurodevelopmental disorders, displayed consistently elevated grooming indices throughout the assay, but transient locomotory activation immediately after placement in the arena. Based on published data identifying FMRP-target transcripts and functional analyses of mutations causing human genetic neurodevelopmental disorders, we propose the following proteins as candidate mediators of excessive repetitive behaviors in FXS: CaMKIIα, NMDA receptor subunits 2A and 2B, NLGN3, and SHANK3. Together, these fly-mutant phenotypes and mechanistic insights provide starting points for drug discovery to identify compounds that reduce dysfunctional repetitive behaviors.

Genome-wide divergence among invasive populations of Aedes aegypti in California.

BACKGROUND: In the summer of 2013, Aedes aegypti Linnaeus was first detected in three cities in central California (Clovis, Madera and Menlo Park). It has now been detected in multiple locations in central and southern CA as far south as San Diego and Imperial Counties. A number of published reports suggest that CA populations have been established from multiple independent introductions. RESULTS: Here we report the first population genomics analyses of Ae. aegypti based on individual, field collected whole genome sequences. We analyzed 46 Ae. aegypti genomes to establish genetic relationships among populations from sites in California, Florida and South Africa. Based on 4.65 million high quality biallelic SNPs, we identified 3 major genetic clusters within California; one that includes all sample sites in the southern part of the state (South of Tehachapi mountain range) plus the town of Exeter in central California and two additional clusters in central California. CONCLUSIONS: A lack of concordance between mitochondrial and nuclear genealogies suggests that the three founding populations were polymorphic for two main mitochondrial haplotypes prior to being introduced to California. One of these has been lost in the Clovis populations, possibly by a founder effect. Genome-wide comparisons indicate extensive differentiation between genetic clusters. Our observations support recent introductions of Ae. aegypti into California from multiple, genetically diverged source populations. Our data reveal signs of hybridization among diverged populations within CA. Genetic markers identified in this study will be of great value in pursuing classical population genetic studies which require larger sample sizes.

Characteristics of tiger moth (Erebidae: Arctiinae) anti-bat sounds can be predicted from tymbal morphology.

BACKGROUND: Acoustic signals are used by many animals to transmit information. Variation in the acoustic characteristics of these signals often covaries with morphology and can relay information about an individual's fitness, sex, species, and/or other characteristics important for both mating and defense. Tiger moths (Lepidoptera: Erebidae: Arctiinae) use modified cuticular plates called "tymbal organs" to produce ultrasonic clicks which can aposematically signal their toxicity, mimic the signals of other species, or, in some cases, disrupt bat echolocation. The morphology of the tymbal organs and the sounds they produce vary greatly between species, but it is unclear how the variation in morphology gives rise to the variation in acoustic characteristics. This is the first study to determine how the morphological features of tymbals can predict the acoustic characteristics of the signals they produce. RESULTS: We show that the number of striations on the tymbal surface (historically known as "microtymbals") and, to a lesser extent, the ratio of the projected surface area of the tymbal to that of the thorax have a strong, positive correlation with the number of clicks a moth produces per unit time. We also found that some clades have significantly different regression coefficients, and thus the relationship between microtymbals and click rate is also dependent on the shared ancestry of different species. CONCLUSIONS: Our predictive model allows the click rates of moths to be estimated using preserved material (e.g., from museums) in cases where live specimens are unavailable. This has the potential to greatly accelerate our understanding of the distribution of sound production and acoustic anti-bat strategies employed by tiger moths. Such knowledge will generate new insights into the evolutionary history of tiger moth anti-predator defenses on a global scale.

Active acoustic interference elicits echolocation changes in heterospecific bats.

Echolocating bats often forage in the presence of both conspecific and heterospecific individuals, which have the potential to produce acoustic interference. Recent studies have shown that at least one bat species, the Brazilian free-tailed bat (Tadarida brasiliensis), produces specialized social signals that disrupt the sonar of conspecific competitors. We herein discuss the differences between passive and active jamming signals and test whether heterospecific jamming occurs in species overlapping spatiotemporally, as well as whether such interference elicits a jamming avoidance response. We compare the capture rates of tethered moths and the echolocation parameters of big brown bats (Eptesicus fuscus) challenged with the playback of the jamming signal normally produced by Brazilian free-tailed bats and playback of deconstructed versions of this signal. There were no differences in the capture rates of targets with and without the jamming signal, although significant changes in both spectral and temporal features of the bats' echolocation were observed. These changes are consistent with improvements of the signal-to-noise ratio in the presence of acoustic interference. Accordingly, we propose to expand the traditional definition of the jamming avoidance response, stating that echolocation changes in response to interference should decrease similarity between the two signals, to include any change that increases the ability to separate returning echoes from active jamming stimuli originating from conspecific and heterospecific organisms. Flexibility in echolocation is an important characteristic for overcoming various forms of acoustic interference and may serve a purpose in interspecific interactions as well as intraspecific ones.

Causal mechanisms of soil organic matter decomposition: deconstructing salinity and flooding impacts in coastal wetlands.

Coastal wetlands significantly contribute to global carbon storage potential. Sea-level rise and other climate-change-induced disturbances threaten coastal wetland sustainability and carbon storage capacity. It is critical that we understand the mechanisms controlling wetland carbon loss so that we can predict and manage these resources in anticipation of climate change. However, our current understanding of the mechanisms that control soil organic matter decomposition, in particular the impacts of elevated salinity, are limited, and literature reports are contradictory. In an attempt to improve our understanding of these complex processes, we measured root and rhizome decomposition and developed a causal model to identify and quantify the mechanisms that influence soil organic matter decomposition in coastal wetlands that are impacted by sea-level rise. We identified three causal pathways: (1) a direct pathway representing the effects of flooding on soil moisture, (2) a direct pathway representing the effects of salinity on decomposer microbial communities and soil biogeochemistry, and (3) an indirect pathway representing the effects of salinity on litter quality through changes in plant community composition over time. We used this model to test the effects of alternate scenarios on the response of tidal freshwater forested wetlands and oligohaline marshes to short- and long-term climate-induced disturbances of flooding and salinity. In tidal freshwater forested wetlands, the model predicted less decomposition in response to drought, hurricane salinity pulsing, and long-term sea-level rise. In contrast, in the oligohaline marsh, the model predicted no change in response to drought and sea-level rise, and increased decomposition following a hurricane salinity pulse. Our results show that it is critical to consider the temporal scale of disturbance and the magnitude of exposure when assessing the effects of salinity intrusion on carbon mineralization in coastal wetlands. Here, we identify three causal mechanisms that can reconcile disparities between long-term and short-term salinity impacts on organic matter decomposition.

How moths escape bats: predicting outcomes of predator-prey interactions.

What determines whether fleeing prey escape from attacking predators? To answer this question, biologists have developed mathematical models that incorporate attack geometries, pursuit and escape trajectories, and kinematics of predator and prey. These models have rarely been tested using data from actual predator-prey encounters. To address this problem, we recorded multi-camera infrared videography of bat-insect interactions in a large outdoor enclosure. We documented 235 attacks by four Myotis volans bats on a variety of moths. Bat and moth flight trajectories from 50 high-quality attacks were reconstructed in 3-D. Despite having higher maximum velocity, deceleration and overall turning ability, bats only captured evasive prey in 69 of 184 attacks (37.5%); bats captured nearly all moths not evading attack (50 of 51; 98%). Logistic regression indicated that prey radial acceleration and escape angle were the most important predictors of escape success (44 of 50 attacks correctly classified; 88%). We found partial support for the turning gambit mathematical model; however, it underestimated the escape threshold by 25% of prey velocity and did not account for prey escape angle. Whereas most prey escaping strikes flee away from predators, moths typically escaped chasing bats by turning with high radial acceleration toward 'safety zones' that flank the predator. This strategy may be widespread in prey engaged in chases. Based on these findings, we developed a novel geometrical model of predation. We discuss implications of this model for the co-evolution of predator and prey kinematics and pursuit and escape strategies.

Rapid host switching of Wolbachia and even more rapid turnover of their phages and incompatibility-causing loci.

About half of all insect species carry maternally inherited Wolbachia alphaproteobacteria, making Wolbachia the most common endosymbionts known in nature. Often Wolbachia spread to high frequencies within populations due to cytoplasmic incompatibility (CI), a Wolbachia-induced sperm modification caused by prophage-associated genes (cifs) that kill embryos without Wolbachia. Several Wolbachia variants also block viruses, including wMel from Drosophila melanogaster when transinfected into the mosquito Aedes aegypti. CI enables the establishment and stable maintenance of pathogen-blocking wMel in natural Ae. aegypti populations. These transinfections are reducing dengue disease incidence on multiple continents. While it has long been known that closely related Wolbachia occupy distantly related hosts, the timing of Wolbachia host switching and molecular evolution has not been widely quantified. We provide a new, conservative calibration for Wolbachia chronograms based on examples of co-divergence of Wolbachia and their insect hosts. Synthesizing publicly available and new genomic data, we use our calibration to demonstrate that wMel-like variants separated by only about 370,000 years have naturally colonized holometabolous dipteran and hymenopteran insects that diverged approximately 350 million years ago. Data from Wolbachia variants closely related to those currently dominant in D. melanogaster and D. simulans illustrate that cifs are rapidly acquired and lost among Wolbachia genomes, on a time scale of 104-105 years. This turnover occurs with and without the Wovirus prophages that contain them, with closely related cifs found in distantly related phages and distantly related cifs found in closely related phages. We present evidence for purifying selection on CI rescue function and on particular Cif protein domains. Our results quantify the tempo and mode of rapid host switching and horizontal gene transfer that underlie the spread and diversity of Wolbachia sampled from diverse host species. The wMel variants we highlight from hosts in different climates may offer new options for broadening Wolbachia-based biocontrol of diseases and pests.

Sound strategies: the 65-million-year-old battle between bats and insects.

The intimate details regarding the coevolution of bats and moths have been elucidated over the past 50 years. The bat-moth story began with the evolution of bat sonar, an exquisite ultrasonic system for tracking prey through the night sky. Moths countered with ears tuned to the high frequencies of bat echolocation and with evasive action through directed turns, loops, spirals, drops, and power dives. Some bat species responded by moving the frequency and intensity of their echolocation cries away from the peak sensitivity of moth ears, and the arms race was on. Tiger moths countered by producing anti-bat sounds. Do the sounds advertise moth toxicity, similar to the bright coloration of butterflies; do they startle the bat, giving the moth a momentary advantage in their aerobatic battle; or do they jam the sonar of the bat? The answer is yes. They do all and more in different situations and in different species. Any insect that flies at night must deal with bat predation. Beetles, mantids, true crickets, mole crickets, katydids, green lacewings, and locusts have anti-bat strategies, and we have just scratched the surface. In an exciting new twist, researchers are taking the technologies developed in the laboratory back into the field, where they are poised to appreciate the full richness of this remarkable predator-prey interaction.

Twenty-Seven Year Response of South Carolina Coastal Plain Forests Affected by Hurricane Hugo.

In 1989, Hurricane Hugo inflicted catastrophic damage on approximately 1.8 million ha of forested land in South Carolina. The purpose of this study was to monitor species compositional shifts and structural changes in several forest types following the hurricane's disturbance. The immediate consequences of hurricane damage are well documented, but there are few studies based on the long-term compositional and structural changes that may result from hurricane disturbance, especially in temperate forest ecosystems. Forty-two forested plots were monitored within four study areas that received varying degrees of hurricane damage. Inventories included species, damage class, tree diameter, and regeneration. The objectives of this study were (1) to compare the recovery speed of wetland forests (e.g., bottomland hardwood swamps and cypress-tupelo swamps) to that of upland pine and hardwood forests; (2) to discover how the degree of hurricane damage can affect the timing and the pattern of forest recovery in the coastal plain; and (3) to compare individual species response patterns across different forest types and at different levels of initial damage. Over the 27-year period following the hurricane, successional pathways have been variable among plots of different forest types and intensity of initial disturbance. We have observed an expected increase in basal area (BA) following the disturbance. Sapling populations in many species have increased dramatically, and some of these populations have begun to thin in recent years. In several forest types, loblolly pine (Pinus taeda L.-not a predominant species in these sites prior to the hurricane) responded quickly and overtook some dominant species in BA and tree/sapling abundance. Several other species that were not a major component of the tree strata (wax myrtle [Morella cerifera (L.) Small], green ash [Fraxinus pennsylvanica Marsh.], and the invasive Chinese tallow [Triadica sebifera (L.) Small]) showed a large increase in sapling population. Overall, recovery speed and species resilience were specific to forest types and damage severity. The intensity and frequency of hurricanes may increase in the future as sea surface temperatures rise. Understanding how coastal forests respond to major hurricanes in the short-term and the long-term will aid us in preparing for future hurricanes and for potential changes in disturbance regimes.

Sonar jamming in the field: effectiveness and behavior of a unique prey defense.

Bats and insects provide a model system for integrating our understanding of predator-prey ecology, animal behavior and neurophysiology. Previous field studies of bat-insect interactions have been limited by the technological challenges involved with studying nocturnal, volant animals that use ultrasound and engage in battles that frequently last a fraction of a second. We overcame these challenges using a robust field methodology that included multiple infrared cameras calibrated for three-dimensional reconstruction of bat and moth flight trajectories and four ultrasonic microphones that provided a spatial component to audio recordings. Our objectives were to document bat-moth interactions in a natural setting and to test the effectiveness of a unique prey defense - sonar jamming. We tested the effect of sonar jamming by comparing the results of interactions between bats and Grote's tiger moth, Bertholdia trigona, with their sound-producing organs either intact or ablated. Jamming was highly effective, with bats capturing more than 10 times as many silenced moths as clicking moths. Moths frequently combined their acoustic defense with two separate evasive maneuvers: flying away from the bat and diving. Diving decreased bat capture success for both clicking and silenced moths, while flying away did not. The diving showed a strong directional component, a first for insect defensive maneuvers. We discuss the timing of B. trigona defensive maneuvers - which differs from that of other moths - in the context of moth auditory neuroethology. Studying bat-insect interactions in their natural environment provides valuable information that complements work conducted in more controlled settings.

cifB-transcript levels largely explain cytoplasmic incompatibility variation across divergent Wolbachia.

Divergent hosts often associate with intracellular microbes that influence their fitness. Maternally transmitted Wolbachia bacteria are the most common of these endosymbionts, due largely to cytoplasmic incompatibility (CI) that kills uninfected embryos fertilized by Wolbachia-infected males. Closely related infections in females rescue CI, providing a relative fitness advantage that drives Wolbachia to high frequencies. One prophage-associated gene (cifA) governs rescue, and two contribute to CI (cifA and cifB), but CI strength ranges from very strong to very weak for unknown reasons. Here, we investigate CI-strength variation and its mechanistic underpinnings in a phylogenetic context across 20 million years (MY) of Wolbachia evolution in Drosophila hosts diverged up to 50 MY. These Wolbachia encode diverse Cif proteins (100% to 7.4% pairwise similarity), and AlphaFold structural analyses suggest that CifB sequence similarities do not predict structural similarities. We demonstrate that cifB-transcript levels in testes explain CI strength across all but two focal systems. Despite phylogenetic discordance among cifs and the bulk of the Wolbachia genome, closely related Wolbachia tend to cause similar CI strengths and transcribe cifB at similar levels. This indicates that other non-cif regions of the Wolbachia genome modulate cif-transcript levels. CI strength also increases with the length of the host's larval life stage, presumably due to prolonged cif action. Our findings reveal that cifB-transcript levels largely explain CI strength, while highlighting other covariates. Elucidating CI's mechanism contributes to our understanding of Wolbachia spread in natural systems and to improving the efficacy of CI-based biocontrol of arboviruses and agricultural pests globally.

Temperature effects on cellular host-microbe interactions explain continent-wide endosymbiont prevalence.

Endosymbioses influence host physiology, reproduction, and fitness, but these relationships require efficient microbe transmission between host generations to persist. Maternally transmitted Wolbachia are the most common known endosymbionts,1 but their frequencies vary widely within and among host populations for unknown reasons.2,3 Here, we integrate genomic, cellular, and phenotypic analyses with mathematical models to provide an unexpectedly simple explanation for global wMel Wolbachia prevalence in Drosophila melanogaster. Cooling temperatures decrease wMel cellular abundance at a key stage of host oogenesis, producing temperature-dependent variation in maternal transmission that plausibly explains latitudinal clines of wMel frequencies on multiple continents. wMel sampled from a temperate climate targets the germline more efficiently in the cold than a recently differentiated tropical variant (∼2,200 years ago), indicative of rapid wMel adaptation to climate. Genomic analyses identify a very narrow list of wMel alleles-most notably, a derived stop codon in the major Wolbachia surface protein WspB-that underlie thermal sensitivity of cellular Wolbachia abundance and covary with temperature globally. Decoupling temperate wMel and host genomes further reduces transmission in the cold, a pattern that is characteristic of host-microbe co-adaptation to a temperate climate. Complex interactions among Wolbachia, hosts, and the environment (GxGxE) mediate wMel cellular abundance and maternal transmission, implicating temperature as a key determinant of Wolbachia spread and equilibrium frequencies, in conjunction with Wolbachia effects on host fitness and reproduction.4,5 Our results motivate the strategic use of locally selected wMel variants for Wolbachia-based biocontrol efforts, which protect millions of individuals from arboviruses that cause human disease.6.

How do tiger moths jam bat sonar?

The tiger moth Bertholdia trigona is the only animal in nature known to defend itself by jamming the sonar of its predators - bats. In this study we analyzed the three-dimensional flight paths and echolocation behavior of big brown bats (Eptesicus fuscus) attacking B. trigona in a flight room over seven consecutive nights to determine the acoustic mechanism of the sonar-jamming defense. Three mechanisms have been proposed: (1) the phantom echo hypothesis, which states that bats misinterpret moth clicks as echoes; (2) the ranging interference hypothesis, which states that moth clicks degrade the bats' precision in determining target distance; and (3) the masking hypothesis, which states that moth clicks mask the moth echoes entirely, making the moth temporarily invisible. On nights one and two of the experiment, the bats appeared startled by the clicks; however, on nights three through seven, the bats frequently missed their prey by a distance predicted by the ranging interference hypothesis (∼15-20 cm). Three-dimensional simulations show that bats did not avoid phantom targets, and the bats' ability to track clicking prey contradicts the predictions of the masking hypothesis. The moth clicks also forced the bats to reverse their stereotyped pattern of echolocation emissions during attack, even while bats continued pursuit of the moths. This likely further hinders the bats' ability to track prey. These results have implications for the evolution of sonar jamming in tiger moths, and we suggest evolutionary pathways by which sonar jamming may have evolved from other tiger moth defense mechanisms.

Arabidopsis CALMODULIN-LIKE 38 Regulates Hypoxia-Induced Autophagy of SUPPRESSOR OF GENE SILENCING 3 Bodies.

During the energy crisis associated with submergence stress, plants restrict mRNA translation and rapidly accumulate stress granules that act as storage hubs for arrested mRNA complexes. One of the proteins associated with hypoxia-induced stress granules in Arabidopsis thaliana is the calcium-sensor protein CALMODULIN-LIKE 38 (CML38). Here, we show that SUPPRESSOR OF GENE SILENCING 3 (SGS3) is a CML38-binding protein, and that SGS3 and CML38 co-localize within hypoxia-induced RNA stress granule-like structures. Hypoxia-induced SGS3 granules are subject to turnover by autophagy, and this requires both CML38 as well as the AAA+-ATPase CELL DIVISION CYCLE 48A (CDC48A). CML38 also interacts directly with CDC48A, and CML38 recruits CDC48A to CML38 granules in planta. Together, this work demonstrates that SGS3 associates with stress granule-like structures during hypoxia stress that are subject to degradation by CML38 and CDC48-dependent autophagy. Further, the work identifies direct regulatory targets for the hypoxia calcium-sensor CML38, and suggest that CML38 association with stress granules and associated regulation of autophagy may be part of the RNA regulatory program during hypoxia stress.

Wolbachia in the spittlebug Prosapia ignipectus: Variable infection frequencies, but no apparent effect on host reproductive isolation.

Animals serve as hosts for complex communities of microorganisms, including endosymbionts that live inside their cells. Wolbachia bacteria are perhaps the most common endosymbionts, manipulating host reproduction to propagate. Many Wolbachia cause cytoplasmic incompatibility (CI), which results in reduced egg hatch when uninfected females mate with infected males. Wolbachia that cause intense CI spread to high and relatively stable frequencies, while strains that cause weak or no CI tend to persist at intermediate, often variable, frequencies. Wolbachia could also contribute to host reproductive isolation (RI), although current support for such contributions is limited to a few systems. To test for Wolbachia frequency variation and effects on host RI, we sampled several local Prosapia ignipectus (Fitch) (Hemiptera: Cercopidae) spittlebug populations in the northeastern United States over two years, including closely juxtaposed Maine populations with different monomorphic color forms, "black" and "lined." We discovered a group-B Wolbachia (wPig) infecting P. ignipectus that diverged from group-A Wolbachia-like model wMel and wRi strains in Drosophila-6 to 46 MYA. Populations of the sister species Prosapia bicincta (Say) from Hawaii and Florida are uninfected, suggesting that P. ignipectus acquired wPig after their initial divergence. wPig frequencies were generally high and variable among sites and between years. While phenotyping wPig effects on host reproduction is not currently feasible, the wPig genome contains three divergent sets of CI loci, consistent with high wPig frequencies. Finally, Maine monomorphic black and monomorphic lined populations of P. ignipectus share both wPig and mtDNA haplotypes, implying no apparent effect of wPig on the maintenance of this morphological contact zone. We hypothesize P. ignipectus acquired wPig horizontally as observed for many Drosophila species, and that significant CI and variable transmission produce high but variable wPig frequencies.

Population genomics of Drosophila suzukii reveal longitudinal population structure and signals of migrations in and out of the continental United States.

Drosophila suzukii, or spotted-wing drosophila, is now an established pest in many parts of the world, causing significant damage to numerous fruit crop industries. Native to East Asia, D. suzukii infestations started in the United States a decade ago, occupying a wide range of climates. To better understand invasion ecology of this pest, knowledge of past migration events, population structure, and genetic diversity is needed. In this study, we sequenced whole genomes of 237 individual flies collected across the continental United States, as well as several sites in Europe, Brazil, and Asia, to identify and analyze hundreds of thousands of genetic markers. We observed strong population structure between Western and Eastern US populations, but no evidence of any population structure between different latitudes within the continental United States, suggesting that there are no broad-scale adaptations occurring in response to differences in winter climates. We detect admixture from Hawaii to the Western United States and from the Eastern United States to Europe, in agreement with previously identified introduction routes inferred from microsatellite analysis. We also detect potential signals of admixture from the Western United States back to Asia, which could have important implications for shipping and quarantine policies for exported agriculture. We anticipate this large genomic dataset will spur future research into the genomic adaptations underlying D. suzukii pest activity and development of novel control methods for this agricultural pest.