BeeDNA: microfluidic environmental DNA metabarcoding as a tool for connecting plant and pollinator communities

Pollinators are imperiled by global declines that can impair plant reproduction, erode essential ecosystem services and resilience, and drive economic losses. Monitoring pollinator biodiversity trends is key for adaptive conservation and management, but conventional surveys are often costly, time consuming, and require taxonomic expertise. Environmental DNA (eDNA) metabarcoding surveys are booming due to their rapidity, non-invasiveness, and cost efficiency. Microfluidic technology allows multiple primer sets from different markers to be used in eDNA metabarcoding for more comprehensive species inventories whilst minimizing biases associated with individual primer sets. We evaluated microfluidic eDNA metabarcoding for pollinator community monitoring by introducing a bumblebee colony to a greenhouse flower assemblage and sampling natural flower plots. We collected nectar draws, flower swabs, or whole flower heads from four flowering species, including two occurring in both the greenhouse and field. Samples were processed using two eDNA isolation protocols before amplification with 15 primer sets for two markers (COI and 16S). Microfluidic eDNA metabarcoding detected the target bumblebee and greenhouse insects as well as common regional arthropods. Pollinator detection was maximized using whole flower heads preserved in ATL buffer and extracted with a modified Qiagen DNeasy protocol for amplification with COI primers. eDNA surveillance could enhance pollinator assessment by detecting protected and endangered species and being more applicable to remote, inaccessible locations, whilst reducing survey time, effort, and expense. Microfluidic eDNA metabarcoding requires optimization but shows promise in revealing complex networks underpinning critical ecosystem functions and services, enabling more accurate assessments of ecosystem resilience.

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Efficiency Comparison of Environmental DNA Metabarcoding of Freshwater Fishes according to Filters, Extraction Kits, Primer Sets and PCR Methods

Korean Journal of Ecology and Environment ◽

10.11614/ksl.2021.54.3.199 ◽

2021 ◽

Vol 54 (3) ◽

pp. 199-208

Author(s):

Keun-Sik Kim ◽

Keun-Yong Kim ◽

Ju-Duk Yoon

Keyword(s):

Environmental Dna ◽

Freshwater Fishes ◽

Dna Metabarcoding ◽

Efficiency Comparison ◽

Primer Sets

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The spatial network structure of intertidal meiofaunal communities derived from environmental DNA metabarcoding surveys in Northwest Iberia

10.1101/2021.03.16.435605 ◽

2021 ◽

Author(s):

Bruno Bellisario ◽

Maria Fais ◽

Sofia Duarte ◽

Pedro Vieira ◽

Carlos Canchaya ◽

...

Keyword(s):

Biological Diversity ◽

Scale Up ◽

Environmental Dna ◽

Taxonomic Diversity ◽

Ecosystem Functions ◽

Individual Species ◽

Ecological Communities ◽

Spatial Networks ◽

Dna Metabarcoding ◽

High Degree

The identification of the patterns and mechanisms behind species distribution is one of the major challenges in ecology, having also important outcomes for the conservation and management of ecosystems. This is especially true for those components of biodiversity providing essential ecosystem functions and for which standard surveys may underestimate their real taxonomic diversity due to their high degree of cryptic diversity and inherent diagnosis difficulties, such as meiofaunal communities. Environmental DNA (eDNA) metabarcoding may provide a fast and reliable way to refine and scale-up the characterization of biological diversity in complex environmental samples, allowing to bypass such drawbacks and increase the resolution of biodiversity estimates. Moreover, the possibility of integrating eDNA metabarcoding-derived data with tools and methods rooted in network theory would deepen the knowledge of the structuring processes of ecological communities in ways that cannot be predicted from studying individual species/communities in isolation. Here, a sediment eDNA metabarcoding of mitochondrial cytochrome c oxidase I (COI) and the nuclear hypervariable V4 region of the 18S rDNA (18S) was used to reconstruct the spatial networks of intertidal meiofaunal OTUs from three estuaries of North-Western Iberian Peninsula. Null models were used to identify the role of environmental and spatial constraints on the structure of COI- and 18S-derived spatial networks and to characterize the macroecological features of surveyed phyla. Our results show the feasibility of eDNA metabarcoding, not only to capture a fair amount of diversity hard to detect with standard surveys procedures, but also to identify hierarchical spatial structures in intertidal meiofaunal assemblages. This suggests that exclusivity of occurrence rather than pervasiveness appears to be the norm in meiofaunal organisms and that niche-based processes predominantly drive the spatial aggregation and contemporary distribution of meiofaunal phyla within the system.

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COI metabarcoding primer choice affects richness and recovery of indicator taxa in freshwater systems

10.1101/572628 ◽

2019 ◽

Cited By ~ 2

Author(s):

Mehrdad Hajibabaei ◽

Teresita M. Porter ◽

Michael Wright ◽

Josip Rudar

Keyword(s):

Beta Diversity ◽

High Throughput Sequencing ◽

Environmental Dna ◽

Sequencing Technology ◽

Indicator Taxa ◽

Combined Use ◽

Dna Metabarcoding ◽

Primer Sets ◽

Major Attention ◽

Biodiversity Analysis

AbstractDNA-based biodiversity analysis has gained major attention due to the use of high throughput sequencing technology in approaches such as mixed community or environmental DNA metabarcoding. Many cytochrome c oxidase subunit I (COI) primer sets are now available for such work. The purpose of this study is to look at how COI primer choice affects the recovery of arthropod richness, beta diversity, and recovery of site indicator taxa in benthos kick-net samples typically used in freshwater biomonitoring. We examine 6 commonly used COI primer sets, on samples collected from 6 freshwater sites. Richness is sensitive to primer choice and the combined use of additional multiple COI amplicons recovers higher richness. Thus, to recover maximum richness, multiple primer sets should be used with COI metabarcoding. Samples consistently cluster by site regardless of amplicon choice or PCR replicate. Thus, for broadscale community analyses, overall beta diversity patterns are robust to COI marker choice. Additionally, the recovery of traditional freshwater bioindicator assemblages such as Ephemeroptera, Trichoptera, Plectoptera, and Diptera may not fully capture the diversity of broadscale arthropod site indicators that can be recovered from COI metabarcoding. Based on these results, studies that use different COI amplicons may not be directly comparable. This work will help future biodiversity and biomonitoring studies develop not just standardized, but optimized workflows that maximize taxon-detection or order taxa along gradients.

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COMPARATIVE STUDY BETWEEN FISH COLLECTION BY NATIONAL CENSUS ON RIVER ENVIRONMENT AND ENVIRONMENTAL DNA METABARCODING

Journal of Japan Society of Civil Engineers Ser B1 (Hydraulic Engineering) ◽

10.2208/jscejhe.74.5_i_415 ◽

2018 ◽

Vol 74 (5) ◽

pp. I_415-I_420 ◽

Cited By ~ 1

Author(s):

Yoshihisa AKAMATSU ◽

Takayoshi TSUZUKI ◽

Ryota YOKOYAMA ◽

Yayoi FUNAHASHI ◽

Munehiro OHTA ◽

...

Keyword(s):

Comparative Study ◽

Environmental Dna ◽

Dna Metabarcoding ◽

Fish Collection

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Environmental DNA for functional diversity

10.1093/oso/9780198767220.003.0010 ◽

2018 ◽

Cited By ~ 1

Author(s):

Pierre Taberlet ◽

Aurélie Bonin ◽

Lucie Zinger ◽

Eric Coissac

Keyword(s):

Functional Groups ◽

Functional Diversity ◽

Environmental Dna ◽

Soil Organisms ◽

Meaningful Information ◽

Dna Metabarcoding ◽

Pros And Cons ◽

Target Community

Chapter 10 “Environmental DNA for functional diversity” discusses the potential of environmental DNA to assess functional diversity. It first focuses on DNA metabarcoding and discusses the extent to which this approach can be used and/or optimized to retrieve meaningful information on the functions of the target community. This knowledge usually involves coarsely defined functional groups (e.g., woody, leguminous, graminoid plants; shredders or decomposer soil organisms; pathogenicity or decomposition role of certain microorganisms). Chapter 10 then introduces metagenomics and metatranscriptomics approaches, their advantages, but also the challenges and solutions to appropriately sampling, sequencing these complex DNA/RNA populations. Chapter 10 finally presents several strategies and software to analyze metagenomes/metatranscriptomes, and discusses their pros and cons.

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Environmental DNA

10.1093/oso/9780198767220.001.0001 ◽

2018 ◽

Cited By ~ 137

Author(s):

Pierre Taberlet ◽

Aurélie Bonin ◽

Lucie Zinger ◽

Eric Coissac

Keyword(s):

Graduate Students ◽

Feeding Habits ◽

Environmental Dna ◽

Research Field ◽

Background Information ◽

Ecological Processes ◽

Biodiversity Research ◽

Dna Metabarcoding ◽

Standard Information

Environmental DNA (eDNA), i.e. DNA released in the environment by any living form, represents a formidable opportunity to gather high-throughput and standard information on the distribution or feeding habits of species. It has therefore great potential for applications in ecology and biodiversity management. However, this research field is fast-moving, involves different areas of expertise and currently lacks standard approaches, which calls for an up-to-date and comprehensive synthesis. Environmental DNA for biodiversity research and monitoring covers current methods based on eDNA, with a particular focus on “eDNA metabarcoding”. Intended for scientists and managers, it provides the background information to allow the design of sound experiments. It revisits all steps necessary to produce high-quality metabarcoding data such as sampling, metabarcode design, optimization of PCR and sequencing protocols, as well as analysis of large sequencing datasets. All these different steps are presented by discussing the potential and current challenges of eDNA-based approaches to infer parameters on biodiversity or ecological processes. The last chapters of this book review how DNA metabarcoding has been used so far to unravel novel patterns of diversity in space and time, to detect particular species, and to answer new ecological questions in various ecosystems and for various organisms. Environmental DNA for biodiversity research and monitoring constitutes an essential reading for all graduate students, researchers and practitioners who do not have a strong background in molecular genetics and who are willing to use eDNA approaches in ecology and biomonitoring.

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