skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: High-throughput sequencing for community analysis: the promise of DNA barcoding to uncover diversity, relatedness, abundances and interactions in spider communities
Abstract Large-scale studies on community ecology are highly desirable but often difficult to accomplish due to the considerable investment of time, labor and, money required to characterize richness, abundance, relatedness, and interactions. Nonetheless, such large-scale perspectives are necessary for understanding the composition, dynamics, and resilience of biological communities. Small invertebrates play a central role in ecosystems, occupying critical positions in the food web and performing a broad variety of ecological functions. However, it has been particularly difficult to adequately characterize communities of these animals because of their exceptionally high diversity and abundance. Spiders in particular fulfill key roles as both predator and prey in terrestrial food webs and are hence an important focus of ecological studies. In recent years, large-scale community analyses have benefitted tremendously from advances in DNA barcoding technology. High-throughput sequencing (HTS), particularly DNA metabarcoding, enables community-wide analyses of diversity and interactions at unprecedented scales and at a fraction of the cost that was previously possible. Here, we review the current state of the application of these technologies to the analysis of spider communities. We discuss amplicon-based DNA barcoding and metabarcoding for the analysis of community diversity and molecular gut content analysis for assessing predator-prey relationships. We also highlight applications of the third generation sequencing technology for long read and portable DNA barcoding. We then address the development of theoretical frameworks for community-level studies, and finally highlight critical gaps and future directions for DNA analysis of spider communities.  more » « less
Award ID(s):
1927510 2104147
PAR ID:
10217560
Author(s) / Creator(s):
; ; ; ; ;
Date Published:
Journal Name:
Development Genes and Evolution
Volume:
230
Issue:
2
ISSN:
0949-944X
Page Range / eLocation ID:
185 to 201
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Anthropogenic influences caused depletion and subsequent recovery of marine predators, but ecological consequences of altered predator abundance are not well understood. Although many methods are used to study predator diets, methodological biases and logistical challenges preclude robust sampling schemes. We aimed to compare two non-invasive methods: metabarcoding scat-derived deoxyribonucleic acid and hard parts analysis of scat for the Northwest Atlantic grey seal ( Halichoerus grypus (Fabricius, 1791)), a species that rebounded after near extirpation. We hypothesized that metabarcoding would detect a greater diversity and frequency of prey, and that notable differences in diet will be detected since prior studies. Grey seal scat samples ( N = 247) were collected between 2018 and 2019 from Monomoy Island, Massachusetts, USA. Metabarcoding detected greater prey richness on average, with more frequent detections of clupeids (Clupeidae) and flatfish (Pleuronectiformes), whereas hard parts analysis more frequently detected phycid hakes ( Urophycis spp. Gill, 1863). Combining methods increased detections of 13 prey taxa, with 32 prey taxa identified overall. Skates (Rajidae), flatfish, clupeids, and sand lance ( Ammodytes spp. Linnaeus, 1758) were top-occurring prey. Our study highlights the importance of using multiple methods to characterize generalist predator diets using non-invasive techniques and suggests grey seal diet has changed since the early 2000s. 
    more » « less
  2. Abstract BackgroundIn light of the current biodiversity crisis, DNA barcoding is developing into an essential tool to quantify state shifts in global ecosystems. Current barcoding protocols often rely on short amplicon sequences, which yield accurate identification of biological entities in a community but provide limited phylogenetic resolution across broad taxonomic scales. However, the phylogenetic structure of communities is an essential component of biodiversity. Consequently, a barcoding approach is required that unites robust taxonomic assignment power and high phylogenetic utility. A possible solution is offered by sequencing long ribosomal DNA (rDNA) amplicons on the MinION platform (Oxford Nanopore Technologies). FindingsUsing a dataset of various animal and plant species, with a focus on arthropods, we assemble a pipeline for long rDNA barcode analysis and introduce a new software (MiniBar) to demultiplex dual indexed Nanopore reads. We find excellent phylogenetic and taxonomic resolution offered by long rDNA sequences across broad taxonomic scales. We highlight the simplicity of our approach by field barcoding with a miniaturized, mobile laboratory in a remote rainforest. We also test the utility of long rDNA amplicons for analysis of community diversity through metabarcoding and find that they recover highly skewed diversity estimates. ConclusionsSequencing dual indexed, long rDNA amplicons on the MinION platform is a straightforward, cost-effective, portable, and universal approach for eukaryote DNA barcoding. Although bulk community analyses using long-amplicon approaches may introduce biases, the long rDNA amplicons approach signifies a powerful tool for enabling the accurate recovery of taxonomic and phylogenetic diversity across biological communities. 
    more » « less
  3. DNA metabarcoding and morphological taxonomic (microscopic) analysis of the gut contents was used to examine diet diversity of seven species of fishes collected from mesopelagic depths (200-1000 m) in the NW Atlantic Ocean Slope Water during Summer 2018 and 2019. Metabarcoding used two gene regions: V9 hypervariable region of nuclear 18S rRNA and mitochondrial cytochrome oxidase I (COI). V9 sequences were classified into 14 invertebrate prey groups, excluding fish due to predator swamping. Ecological network analysis was used to evaluate relative strengths of predator-prey linkages. Multivariate statistical analysis revealed consistently distinct diets of four fish species in 2018 and/or 2019:Argyropelecus aculeatus, Chauliodus sloani, Hygophum hygomii, andSigmops elongatus. Three other species analyzed (Malacosteus niger, Nemichthys scolopaceus, andScopelogadus beanii) showed more variability between sampling years. COI sequences were classified into eight invertebrate prey groups, within which prey species were detected and identified. Considering all predator species together, a total of 77 prey species were detected with a minimum of 1,000 COI sequences, including 22 copepods, 18 euphausiids, and 7 amphipods. Morphological prey counts were classified into seven taxonomic groups, including a gelatinous group comprised of soft-bodied organisms. The ocean twilight zone or is home to exceptional diversity and biomass of marine fish, which are key players in deep sea food webs. This study used integrative morphological-molecular analysis to provide new insights into trophic relationships and sources of productivity for mesopelagic fishes, including identification of key prey species, recognition of the importance of gelatinous prey, and characterization of differences in diet among fish predators in the NW Atlantic Slope Water. 
    more » « less
  4. Abstract Alternative ecological theories make divergent predictions about the relationship between predators and their prey. If predators exert top‐down ecosystem control, increases in predation should diminish prey abundance and could either diminish or enhance community diversity of prey species. However, if bottom‐up ecosystem controls predominate, predator populations should track underlying variation in prey diversity and abundance, which ultimately should reflect available energy. Past research, both across islands and comparing islands with the mainland, has frequently invoked the importance of predation in regulating lizard abundance and diversity, suggesting an important role of top‐down control when predators are present. However, others have posited a stronger role of food limitation, via competition or bottom‐up forces. If top‐down control predominates, then negative correlations between prey abundance and predator occurrence should emerge within and among islands. Using data from eBird, we inferred landscape‐level presence data for bird species on the islands of Jamaica and Hispaniola. By summing occurrence probabilities of all known anole‐predator birds, we estimated total avian predation pressure and combined these estimates with anole community data from a mark‐recapture study that spanned spatial and climatic gradients on both islands. Avian predators and anole lizards were both affected by climate, with total predator occurrence, anole abundance and anole species richness increasing with mean annual temperature. Anole abundance and predator occurrence showed a curvilinear relationship, where abundance and predator occurrence increased together until predator occurrence became sufficiently high that anole abundance was negatively impacted. This indicates that bottom‐up ecosystem controls drive richness of both anoles and their predators, mitigating the negative effects predators might have on their prey, at least until predator occurrence reaches a threshold. We did not detect consistent evidence of predator occurrence reducing anole community richness. These findings support past research showing that islands with more predators tend to have lower prey abundances, but it does not seem that these top‐down forces are strongly limiting species coexistence. Instead, bottom‐up forces linked with climate may be more important drivers of diversity in both lizards and their avian predators on these islands. 
    more » « less
  5. 1. Camera trap technology has galvanized the study of predator-prey ecology in wild animal communities by expanding the scale and diversity of predator-prey interactions that can be analyzed. While observational data from systematic camera arrays have informed inferences on the spatiotemporal outcomes of predator-prey interactions, the capacity for observational studies to identify mechanistic drivers of species interactions is limited. 2. Experimental study designs that utilize camera traps uniquely allow for testing hypothesized mechanisms that drive predator and prey behavior, incorporating environmental realism not possible in the lab while benefiting from the distinct capacity of camera traps to generate large data sets from multiple species with minimal observer interference. However, such pairings of camera traps with experimental methods remain underutilized. 3. We review recent advances in the experimental application of camera traps to investigate fundamental mechanisms underlying predator-prey ecology and present a conceptual guide for designing experimental camera trap studies. 4. Only 9% of camera trap studies on predator-prey ecology in our review mention experimental methods, but the application of experimental approaches is increasing. To illustrate the utility of camera trap-based experiments using a case study, we propose a study design that integrates observational and experimental techniques to test a perennial question in predator-prey ecology: how prey balance foraging and safety, as formalized by the risk allocation hypothesis. We discuss applications of camera trap-based experiments to evaluate the diversity of anthropogenic influences on wildlife communities globally. Finally, we review challenges to conducting experimental camera trap studies. 5. Experimental camera trap studies have already begun to play an important role in understanding the predator-prey ecology of free-living animals, and such methods will become increasingly critical to quantifying drivers of community interactions in a rapidly changing world. We recommend increased application of experimental methods in the study of predator and prey responses to humans, synanthropic and invasive species, and other anthropogenic disturbances. 
    more » « less