Microbes, the 'silent third partners' of bee-angiosperm mutualisms.

While bee-angiosperm mutualisms are widely recognized as foundational partnerships that have shaped the diversity and structure of terrestrial ecosystems, these ancient mutualisms have been underpinned by 'silent third partners': microbes. Here, we propose reframing the canonical bee-angiosperm partnership as a three-way mutualism between bees, microbes, and angiosperms. This new conceptualization casts microbes as active symbionts, processing and protecting pollen-nectar provisions, consolidating nutrients for bee larvae, enhancing floral attractancy, facilitating plant fertilization, and defending bees and plants from pathogens. In exchange, bees and angiosperms provide their microbial associates with food, shelter, and transportation. Such microbial communities represent co-equal partners in tripartite mutualisms with bees and angiosperms, facilitating one of the most important ecological partnerships on land.

A sustainable grower-based method for entomopathogenic nematode production.

Entomopathogenic nematodes in the genera Steinernema and Heterorhabditis, produced through in vitro or in vivo methods, are effective insect biological control agents. In vivo production yields good-quality nematodes, but the costs associated with obtaining insects and labor make this production system have a low economy of scale. Conceivably, if growers can produce their own nematodes, then the cost could be reduced. Grower-based production systems described to-date are not sustainable because they rely on outside sources to obtain or calibrate inoculum. Here, we describe a self-sufficient grower-based system where the grower can produce in-house nematodes after obtaining the initial inoculum from a reliable source. We validated our approach in 2 experiments comparing in vivo nematode production from standard White traps and a grower-based approach using polyacrylamide gel. For both tested species, Steinernema carpocapsae (Weiser) and Heterorhabditis bacteriophora Poinar, the grower-based approach produced equal to or more nematodes than the standard method. For example, when comparing the average yield of S. carpocapsae-infective juveniles per Galleria mellonella cadaver (n = 30), the standard White trap method produced 159,114 ± 9,669, whereas the grower-based approach produced 244,029 ± 16,241. The sustainable system described herein has promise for wide adoption by growers.

Apex Predator Nematodes and Meso-Predator Bacteria Consume Their Basal Insect Prey through Discrete Stages of Chemical Transformations.

Microbial symbiosis drives physiological processes of higher-order systems, including the acquisition and consumption of nutrients that support symbiotic partner reproduction. Metabolic analytics provide new avenues to examine how chemical ecology, or the conversion of existing biomass to new forms, changes over a symbiotic life cycle. We applied these approaches to the nematode Steinernema carpocapsae, its mutualist bacterium, Xenorhabdus nematophila, and the insects they infect. The nematode-bacterium pair infects, kills, and reproduces in an insect until nutrients are depleted. To understand the conversion of insect biomass over time into either nematode or bacterium biomass, we integrated information from trophic, metabolomic, and gene regulation analyses. Trophic analysis established bacteria as meso-predators and primary insect consumers. Nematodes hold a trophic position of 4.6, indicative of an apex predator, consuming bacteria and likely other nematodes. Metabolic changes associated with Galleria mellonella insect bioconversion were assessed using multivariate statistical analyses of metabolomics data sets derived from sampling over an infection time course. Statistically significant, discrete phases were detected, indicating the insect chemical environment changes reproducibly during bioconversion. A novel hierarchical clustering method was designed to probe molecular abundance fluctuation patterns over time, revealing distinct metabolite clusters that exhibit similar abundance shifts across the time course. Composite data suggest bacterial tryptophan and nematode kynurenine pathways are coordinated for reciprocal exchange of tryptophan and NAD+ and for synthesis of intermediates that can have complex effects on bacterial phenotypes and nematode behaviors. Our analysis of pathways and metabolites reveals the chemistry underlying the recycling of organic material during carnivory. IMPORTANCE The processes by which organic life is consumed and reborn in a complex ecosystem were investigated through a multiomics approach applied to the tripartite Xenorhabdus bacterium-Steinernema nematode-Galleria insect symbiosis. Trophic analyses demonstrate the primary consumers of the insect are the bacteria, and the nematode in turn consumes the bacteria. This suggests the Steinernema-Xenorhabdus mutualism is a form of agriculture in which the nematode cultivates the bacterial food sources by inoculating them into insect hosts. Metabolomics analysis revealed a shift in biological material throughout progression of the life cycle: active infection, insect death, and conversion of cadaver tissues into bacterial biomass and nematode tissue. We show that each phase of the life cycle is metabolically distinct, with significant differences including those in the tricarboxylic acid cycle and amino acid pathways. Our findings demonstrate that symbiotic life cycles can be defined by reproducible stage-specific chemical signatures, enhancing our broad understanding of metabolic processes that underpin a three-way symbiosis.

Exosymbiotic microbes within fermented pollen provisions are as important for the development of solitary bees as the pollen itself.

Developing bees derive significant benefits from the microbes present within their guts and fermenting pollen provisions. External microbial symbionts (exosymbionts) associated with larval diets may be particularly important for solitary bees that suffer reduced fitness when denied microbe-colonized pollen.To investigate whether this phenomenon is generalizable across foraging strategy, we examined the effects of exosymbiont presence/absence across two solitary bee species, a pollen specialist and generalist. Larvae from each species were reared on either microbe-rich natural or microbe-deficient sterilized pollen provisions allocated by a female forager belonging to their own species (conspecific-sourced pollen) or that of another species (heterospecific-sourced pollen). Our results reveal that the presence of pollen-associated microbes was critical for the survival of both the generalist and specialist larvae, regardless of whether the pollen was sourced from a conspecific or heterospecific forager.Given the positive effects of exosymbiotic microbes for larval fitness, we then examined if the magnitude of this benefit varied based on whether the microbes were provisioned by a conspecific forager (the mother bee) or a heterospecific forager. In this second study, generalist larvae were reared only on microbe-rich pollen provisions, but importantly, the sources (conspecific versus heterospecific) of the microbes and pollen were experimentally manipulated.Bee fitness metrics indicated that microbial and pollen sourcing both had significant impacts on larval performance, and the effect sizes of each were similar. Moreover, the effects of conspecific-sourced microbes and conspecific-sourced pollen were strongly positive, while that of heterospecific-sourced microbes and heterospecific-sourced pollen, strongly negative.Our findings imply that not only is the presence of exosymbionts critical for both specialist and generalist solitary bees, but more notably, that the composition of the specific microbial community within larval pollen provisions may be as critical for bee development as the composition of the pollen itself.

Drones That Deliver: Pheromone-Based Mating Disruption Deployed via Uncrewed Aerial Vehicles in U.S. Cranberries.

Cranberry fruitworm (Acrobasis vaccinii Riley (Lepidoptera: Pyralidae)) and blackheaded fireworm (Rhopobota naevana Hubner (Lepidoptera: Tortricidae)) threaten cranberry production annually by causing significant fruit damage. Up to four pesticide applications are made each year to control these insects, which are costly to producers and elevate pesticide residues in fruit. Pheromone-based mating disruption technology can provide control of these pests in cranberry production, with the potential to minimize, or eliminate, pesticide applications. In 2016, an uncrewed aerial vehicle (UAV) was investigated to apply a thick paraffin emulsion containing insect sex pheromones. Traditional agricultural equipment is not capable of applying the paraffin emulsion to cranberry beds due to the product's viscous, paste-like consistency. The first objective of this study was to retrofit an UAV (octocopter) with a novel extrusion device that had been engineered to deliver the pheromone-loaded paraffin at regular intervals during flight. The second objective was to confirm adequate distribution of the pheromones by measuring the mating disruption efficacy by monitoring male moth trap catches. The UAV was able to fly autonomously along a prescribed itinerary, deploying the paraffin product uniformly; however, the increased mass of the retrofitted UAV limited flight times to ~12 min. The number of male cranberry fruitworm and blackheaded fireworm moths caught in lure-baited traps were reduced in the paraffin-treated beds compared with untreated beds, indicating adequate distribution of the pheromones. The UAV-applied pheromones concept could be developed into a production scale application method in the future, although issues of battery life and lifting capacity will need to be resolved.

(More than) Hitchhikers through the network: the shared microbiome of bees and flowers.

Growing evidence reveals strong overlap between microbiomes of flowers and bees, suggesting that flowers are hubs of microbial transmission. Whether floral transmission is the main driver of bee microbiome assembly, and whether functional importance of florally sourced microbes shapes bee foraging decisions are intriguing questions that remain unanswered. We suggest that interaction network properties, such as nestedness, connectedness, and modularity, as well as specialization patterns can predict potential transmission routes of microbes between hosts. Yet microbial filtering by plant and bee hosts determines realized microbial niches. Functionally, shared floral microbes can provide benefits for bees by enhancing nutritional quality, detoxification, and disintegration of pollen. Flower microbes can also alter the attractiveness of floral resources. Together, these mechanisms may affect the structure of the flower-bee interaction network.

Microbial Diversity Associated with the Pollen Stores of Captive-Bred Bumble Bee Colonies.

The pollen stores of bumble bees host diverse microbiota that influence overall colony fitness. Yet, the taxonomic identity of these symbiotic microbes is relatively unknown. In this descriptive study, we characterized the microbial community of pollen provisions within captive-bred bumble bee hives obtained from two commercial suppliers located in North America. Findings from 16S rRNA and ITS gene-based analyses revealed that pollen provisions from the captive-bred hives shared several microbial taxa that have been previously detected among wild populations. While diverse microbes across phyla Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Ascomycota were detected in all commercial hives, significant differences were detected at finer-scale taxonomic resolution based on the supplier source. The causative agent of chalkbrood disease in honey bees, Ascosphaera apis, was detected in all hives obtained from one supplier source, although none of the hives showed symptoms of infection. The shared core microbiota across both commercial supplier sources consisted of two ubiquitous bee-associated groups, Lactobacillus and Wickerhamiella/Starmerella clade yeasts that potentially contribute to the beneficial function of the microbiome of bumble bee pollen provisions.

A Rare, Recently Discovered Nematode, Oscheius onirici (Rhabditida: Rhabditidae), Kills Drosophila suzukii (Diptera: Drosophilidae) Within Fruit.

The spotted-wing drosophila, Drosophila suzukii Matsumura, is an exotic species in North America and represents a major threat to fruit production. Efforts to manage D. suzukii have focused primarily on insecticides, but such controls may, at times, be unreliable, given that D. suzukii larvae are often ensconced within fruit. The fruit interior, however, may represent suitable foraging substrates for carnivorous/entomopathogenic nematodes. In preliminary trials, a rare nematode species, Oscheius onirici Torrini et al., was shown to be highly virulent against D. suzukii when the nematodes were applied directly to fly larvae. To address the more important question of whether this nematode would be as virulent when applied to fruit, we set up assays in which blueberries were infested with D. suzukii larvae and then sprayed with O. onirici infective juveniles (IJs). Across two laboratory trials, O. onirici IJs suppressed D. suzukii puparia by 78.2%. Oscheius onirici IJs were able to search effectively within fruit substrates, find the fly larvae therein, and kill the flies before they could pupariate. Oscheius onirici, therefore, may represent a viable new bio-control agent for D. suzukii management and should be field-tested across a broader diversity of cropping systems.

Omnivory in Bees: Elevated Trophic Positions among All Major Bee Families.

As pollen and nectar foragers, bees have long been considered strictly herbivorous. Their pollen provisions, however, are host to abundant microbial communities, which feed on the pollen before and/or while it is consumed by bee larvae. In the process, microbes convert pollen into a complex of plant and microbial components. Since microbes are analogous to metazoan consumers within trophic hierarchies, the pollen-eating microbes are, functionally, herbivores. When bee larvae consume a microbe-rich pollen complex, they ingest proteins from plant and microbial sources and thus should register as omnivores on the trophic "ladder." We tested this hypothesis by examining the isotopic compositions of amino acids extracted from native bees collected in North America over multiple years. We measured bee trophic position across the six major bee families. Our findings indicate that bee trophic identity was consistently and significantly higher than that of strict herbivores, providing the first evidence that omnivory is ubiquitous among bee fauna. Such omnivory suggests that pollen-borne microbes represent an important protein source for larval bees, which introduces new questions as to the link between floral fungicide residues and bee development.

Pollen-borne microbes shape bee fitness.

Teeming within pollen provisions are diverse communities of symbiotic microbes, which provide a variety of benefits to bees. Microbes themselves may represent a major dietary resource for developing bee larvae. Despite their apparent importance in sustaining bee health, evidence linking pollen-borne microbes to larval health is currently lacking. We examined the effects of microbe-deficient diets on the fitness of larval mason bees. In a series of diet manipulations, microbe-rich maternally collected pollen provisions were replaced with increasing fractions of sterilized, microbe-deficient pollen provisions before being fed to developing larvae. Convergent findings from amino acid and fatty acid trophic biomarker analyses revealed that larvae derived a substantial amount of nutrition from microbial prey and occupied a significantly higher trophic position than that of strict herbivores. Larvae feeding on increasingly sterile diets experienced significant adverse effects on growth rates, biomass and survivorship. When completely deprived of pollen-borne microbes, larvae consistently exhibited marked decline in fitness. We conclude that microbes associated with aged pollen provisions are central to bee health, not only as nutritional mutualists, but also as a major dietary component. In an era of global bee decline, the conservation of such bee-microbe interactions may represent an important facet of pollinator protection strategies.

Incidence of Oscheius onirici (Nematoda: Rhabditidae), a potentially entomopathogenic nematode from the marshlands of Wisconsin, USA.

In a search for an entomopathogenic nematode to control cranberry insect pests, three Oscheius populations (Rhabditidae) were recovered through the Galleria-bait method from one sample taken in a wild cranberry marsh in Jackson County, Wisconsin, USA. Morphological studies with light microscopy and scanning electron microscopy, as well as molecular analyses of the near-full-length small subunit rDNA gene, D2/D3 expansion segments of the large subunit rDNA gene, internal transcribed spacer, and mitochondrial cytochrome oxidase subunit 1 (CoxI) genes revealed this as Oscheius onirici, a species recently described from a karst cave soil of central Italy. The species belongs to the dolichura-group and is characterized by its DNA sequences; hermaphroditic reproduction; and males not found. A Bacillus-like bacterium appears to be associated with this nematode based on our microscopic and SEM observations; however its identity and persistent association with the nematode has not been confirmed. Nonetheless, this nematode is capable of infecting and killing the sparganothis fruitworm Sparganothis sulfureana Clemens (Lepidoptera: Tortricidae), the brown-banded cockroach Supella longipalpa Fabricius (Blattodea: Ectobiidae), and the cranberry fruitworm Acrobasis vaccinii Riley (Lepidoptera: Pyralidae), under laboratory conditions, and each in less than 72 hr. The mealworm Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae) and the greater wax moth Galleria mellonella Linnaeus (Lepidoptera: Pyralidae), are also susceptible, but take 3.5 and 5.2 days to die, respectively. This species is a new potential bio-control agent on insects.

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors.

Although solitary bees provide crucial pollination services for wild and managed crops, this species-rich group has been largely overlooked in pesticide regulation studies. The risk of exposure to fungicide residues is likely to be especially high if the spray occurs on, or near host plants while the bees are collecting pollen to provision their nests. For species of Osmia that consume pollen from a select group of plants (oligolecty), the inability to use pollen from non-host plants can increase their risk factor for fungicide-related toxicity. This manuscript describes protocols used to successfully rear oligolectic mason bees, Osmia ribifloris sensu lato, from egg to prepupal stage within cell culture plates under standardized laboratory conditions. The in vitro-reared bees are subsequently used to investigate the effects of fungicide exposure and pollen source on bee fitness. Based on a 2 × 2 fully crossed factorial design, the experiment examines the main and interactive effects of fungicide exposure and pollen source on larval fitness, quantified by prepupal biomass, larval developmental time, and survivorship. A major advantage of this technique is that using in vitro-reared bees reduces natural background variability and allows the simultaneous manipulation of multiple experimental parameters. The described protocol presents a versatile tool for hypotheses testing involving the suite of factors affecting bee health. For conservation efforts to be met with significant, lasting success, such insights into the complex interplay of physiological and environmental factors driving bee declines will prove to be critical.

Variable Isotopic Compositions of Host Plant Populations Preclude Assessment of Aphid Overwintering Sites.

Soybean aphid (Aphis glycines Matsumura) is a pest of soybean in the northern Midwest whose migratory patterns have been difficult to quantify. Improved knowledge of soybean aphid overwintering sites could facilitate the development of control efforts with exponential impacts on aphid densities on a regional scale. In this preliminary study, we explored the utility of variation in stable isotopes of carbon and nitrogen to distinguish soybean aphid overwintering origins. We compared variation in bulk 13C and 15N content in buckthorn (Rhamnus cathartica L.) and soybean aphids in Wisconsin, among known overwintering locations in the northern Midwest. Specifically, we looked for associations between buckthorn and environmental variables that could aid in identifying overwintering habitats. We detected significant evidence of correlation between the bulk 13C and 15N signals of soybean aphids and buckthorn, despite high variability in stable isotope composition within and among buckthorn plants. Further, the 15N signal in buckthorn varied predictably with soil composition. However, lack of sufficient differentiation of geographic areas along axes of isotopic and environmental variation appears to preclude the use of carbon and nitrogen isotopic signals as effective predictors of likely aphid overwintering sites. These preliminary data suggest the need for future work that can further account for variability in 13C and 15N within/among buckthorn plants, and that explores the utility of other stable isotopes in assessing likely aphid overwintering sites.

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health.

Growers often use fungicide sprays during bloom to protect crops against disease, which exposes bees to fungicide residues. Although considered "bee-safe," there is mounting evidence that fungicide residues in pollen are associated with bee declines (for both honey and bumble bee species). While the mechanisms remain relatively unknown, researchers have speculated that bee-microbe symbioses are involved. Microbes play a pivotal role in the preservation and/or processing of pollen, which serves as nutrition for larval bees. By altering the microbial community, it is likely that fungicides disrupt these microbe-mediated services, and thereby compromise bee health. This manuscript describes the protocols used to investigate the indirect mechanism(s) by which fungicides may be causing colony decline. Cage experiments exposing bees to fungicide-treated flowers have already provided the first evidence that fungicides cause profound colony losses in a native bumble bee (Bombus impatiens). Using field-relevant doses of fungicides, a series of experiments have been developed to provide a finer description of microbial community dynamics of fungicide-exposed pollen. Shifts in the structural composition of fungal and bacterial assemblages within the pollen microbiome are investigated by next-generation sequencing and metagenomic analysis. Experiments developed herein have been designed to provide a mechanistic understanding of how fungicides affect the microbiome of pollen-provisions. Ultimately, these findings should shed light on the indirect pathway through which fungicides may be causing colony declines.

Unpacking brown food-webs: Animal trophic identity reflects rampant microbivory.

Detritivory is the dominant trophic paradigm in most terrestrial, aquatic, and marine ecosystems, yet accurate measurement of consumer trophic position within detrital (="brown") food webs has remained unresolved. Measurement of detritivore trophic position is complicated by the fact that detritus is suffused with microbes, creating a detrital complex of living and nonliving biomass. Given that microbes and metazoans are trophic analogues of each other, animals feeding on detrital complexes are ingesting other detritivores (microbes), which should elevate metazoan trophic position and should be rampant within brown food webs. We tested these hypotheses using isotopic (15N) analyses of amino acids extracted from wild and laboratory-cultured consumers. Vertebrate (fish) and invertebrate detritivores (beetles and moths) were reared on detritus, with and without microbial colonization. In the field, detritivorous animal specimens were collected and analyzed to compare trophic identities among laboratory-reared and free-roaming detritivores. When colonized by bacteria or fungi, the trophic positions of detrital complexes increased significantly over time. The magnitude of trophic inflation was mediated by the extent of microbial consumption of detrital substrates. When detrital complexes were fed to vertebrate and invertebrate animals, the consumers registered similar degrees of trophic inflation, albeit one trophic level higher than their diets. The wild-collected detritivore fauna in our study exhibited significantly elevated trophic positions. Our findings suggest that the trophic positions of detrital complexes rise predictably as microbes convert nonliving organic matter into living microbial biomass. Animals consuming such detrital complexes exhibit similar trophic inflation, directly attributable to the assimilation of microbe-derived amino acids. Our data demonstrate that detritivorous microbes elevate metazoan trophic position, suggesting that detritivory among animals is, functionally, omnivory. By quantifying the impacts of microbivory on the trophic positions of detritivorous animals and then tracking how these effects propagate "up" food chains, we reveal the degree to which microbes influence consumer groups within trophic hierarchies. The trophic inflation observed among our field-collected fauna further suggests that microbial proteins represent an immense contribution to metazoan biomass. Collectively, these findings provide an empirical basis to interpret detritivore trophic identity, and further illuminate the magnitude of microbial contributions to food webs.

Intra-trophic isotopic discrimination of 15N/14N for amino acids in autotrophs: Implications for nitrogen dynamics in ecological studies.

The differential discrimination of nitrogen isotopes (15N/14N) within amino acids in consumers and their diets has been routinely used to estimate organismal tropic position (TP). Analogous isotopic discrimination can occur within plants, particularly in organs lacking chloroplasts. Such discrimination likely arises from the catabolic deamination of amino acids, resulting in a numerical elevation of estimated TP, within newly synthesized biomass. To investigate this phenomenon, we examined the 15N/14N of amino acids (δ15 NAA) in spring leaves and flowers from eight deciduous and two annual plants. These plants were classified on the basis of their time of bloom, plants that bloomed when their leaves were absent (Type I) versus plants that bloomed while leaves were already present (Type II). Based on the δ15 NAA values from leaves, both plant types occupied comparable and ecologically realistic mean TPs (=1.0 ± 0.1, mean ± 1σ). However, the estimated TPs of flowers varied significantly (Type I: 2.2 ± 0.2; Type II: 1.0 ± 0.1). We hypothesize that these results can be interpreted by the following sequence of events: (1) Type I floral biomass is synthesized in absence of active photosynthesis; (2) the catabolic deamination of amino acids in particular, leaves behind 15N in the residual pool of amino acids; and (3) the incorporation of these 15N-enriched amino acids within the biomass of Type I flowers results in the numerical elevation of the TPs. In contrast, the actively photosynthesizing Type II leaves energetically sustain the synthesis of Type II flower biomass, precluding any reliance on catabolic deamination of amino acids. Amino acids within Type II flowers are therefore isotopically comparable to the Type II leaves. These findings demonstrate the idiosyncratic nature of the δ15 NAA values within autotrophic organs and have implications for interpreting trophic hierarchies using primary producers and their consumers.

Multi-Species Mating Disruption in Cranberries (Ericales: Ericaceae): Early Evidence Using a Flowable Emulsion.

Pheromone-based mating disruption has proven to be a powerful pest management tactic in many cropping systems. However, in the cranberry system, a viable mating disruption program does not yet exist. There are commercially available pheromones for several of the major pests of cranberries, including the cranberry fruitworm, Acrobasis vaccinii Riley (Lepidoptera: Pyralidae) and blackheaded fireworm, Rhopobota naevana (Hübner) (Lepidoptera: Tortricidae). Previous studies have shown that mating disruption represents a promising approach for R. naevana management although carrier and delivery technologies have remained unresolved. The present study examined the suitability of Specialized Pheromone & Lure Application Technology (SPLAT; ISCA Technologies, Inc., Riverside, CA), a proprietary wax and oil blend, to serve as a pheromone carrier in the cranberry system. In 2013 and 2014, we tested a blend of pheromones targeting A. vaccinii and R. naevana in field-scale, replicated trials. Pheromones were loaded into SPLAT and the resulting "SPLAT BFW CFW" formulation was deployed in commercial cranberry marshes. We compared moth trap-catch counts within SPLAT-treated blocks to those of conventionally managed blocks. In 2013, applications of SPLAT BFW CFW resulted in highly successful disruption of R. naevana and promising, though inconsistent, disruption of A. vaccinii. To improve disruption of A. vaccinii, the pheromone load was increased in 2014, providing 92% and 74% reductions in trap-catch for R. naevana and A. vaccinii, respectively. Importantly, larval infestation rates in SPLAT-treated blocks were lower than those of conventionally managed blocks. These results suggest that a multispecies mating disruption system (SPLAT BFW CFW) may represent an effective pesticide-alternative for serious pests of cranberries.

Flight Synchrony among the Major Moth Pests of Cranberries in the Upper Midwest, USA.

The cranberry fruitworm (Acrobasis vaccinii Riley), sparganothis fruitworm (Sparganothis sulfureana Clemens), and blackheaded fireworm (Rhopobota naevana Hübner) are historically significant pests of cranberries (Vaccinium macrocarpon Aiton) in the Upper Midwest (Wisconsin), USA. Their respective natural histories are well documented but correlations between developmental benchmarks (e.g., larval eclosion) and degree-day accruals are not yet known. Treatment timings are critical to the optimization of any given control tactic, and degree-day accrual facilitates optimization by quantifying the developmental status of pest populations. When key developmental benchmarks in the pest life cycle are linked to degree-days, real-time weather data can be used to predict precise treatment timings. Here, we provide the degree-day accumulations associated with discrete biological events (i.e., initiation of flight and peak flight) for the three most consistent moth pests of cranberries in Wisconsin. Moths were trapped each spring and summer from 2003 to 2011. To characterize flight dynamics and average timing of flight initiation, pheromone-baited trap-catch data were tallied for all three pest species within each of seven growing seasons. These flight dynamics were then associated with the corresponding degree-day accumulations generated using the cranberry plant's developmental thresholds. Finally, models were fit to the data in order to determine the peak flight of each species. The initiation of the spring flight among all three moth species was highly synchronous, aiding in the timing of control tactics; however, there were substantial differences in the timing of peak flight among the moth species. Characterization of the relationship between temperature and pest development allows pest management professionals to target specific life stages, improving the efficacy of any given pest control tactic.

Exploiting genotyping by sequencing to characterize the genomic structure of the American cranberry through high-density linkage mapping.

BACKGROUND: The application of genotyping by sequencing (GBS) approaches, combined with data imputation methodologies, is narrowing the genetic knowledge gap between major and understudied, minor crops. GBS is an excellent tool to characterize the genomic structure of recently domesticated (~200 years) and understudied species, such as cranberry (Vaccinium macrocarpon Ait.), by generating large numbers of markers for genomic studies such as genetic mapping. RESULTS: We identified 10842 potentially mappable single nucleotide polymorphisms (SNPs) in a cranberry pseudo-testcross population wherein 5477 SNPs and 211 short sequence repeats (SSRs) were used to construct a high density linkage map in cranberry of which a total of 4849 markers were mapped. Recombination frequency, linkage disequilibrium (LD), and segregation distortion at the genomic level in the parental and integrated linkage maps were characterized for first time in cranberry. SSR markers, used as the backbone in the map, revealed high collinearity with previously published linkage maps. The 4849 point map consisted of twelve linkage groups spanning 1112 cM, which anchored 2381 nuclear scaffolds accounting for ~13 Mb of the estimated 470 Mb cranberry genome. Bin mapping identified 592 and 672 unique bins in the parentals and a total of 1676 unique marker positions in the integrated map. Synteny analyses comparing the order of anchored cranberry scaffolds to their homologous positions in kiwifruit, grape, and coffee genomes provided initial evidence of homology between cranberry and closely related species. CONCLUSIONS: GBS data was used to rapidly saturate the cranberry genome with markers in a pseudo-testcross population. Collinearity between the present saturated genetic map and previous cranberry SSR maps suggests that the SNP locations represent accurate marker order and chromosome structure of the cranberry genome. SNPs greatly improved current marker genome coverage, which allowed for genome-wide structure investigations such as segregation distortion, recombination, linkage disequilibrium, and synteny analyses. In the future, GBS can be used to accelerate cranberry molecular breeding through QTL mapping and genome-wide association studies (GWAS).