Influence of DNA extraction kits on the fungal DNA metabarcoding for freshwater environmental DNA

Background: Environmental DNA (eDNA) metabarcoding is a rapidly expanding technique for efficient biodiversity monitoring, especially of animals. Recently, the usefulness of aquatic eDNA in monitoring the diversity of both terrestrial and aquatic fungi has been suggested. In eDNA studies, different experimental factors, such as DNA extraction kits or methods, can affect the subsequent analyses and the results of DNA metabarcoding. However, few methodological studies have been carried out on eDNA of fungi, and little is known about how experimental procedures can affect the results of biodiversity analysis. In this study, we focused on the effect of the DNA extraction method on fungal DNA metabarcoding using freshwater samples obtained from rivers and lakes. Methods: DNA was extracted from freshwater samples using the DNeasy PowerSoil kit, which is mainly used to extract microbial DNA from soil, and the DNeasy Blood & Tissue kit, which is commonly used for eDNA studies on animals. We then compared PCR inhibition and fungal DNA metabarcoding results [i.e., operational taxonomic unit (OTU) number and composition] of the extracted samples. Results: No PCR inhibition was detected in any of the samples, and no significant differences in the number of OTUs and OTU compositions were detected between the samples processed using different kits. These results indicate that both DNA extraction kits may provide similar diversity results for the river and lake samples evaluated in this study. Therefore, it may be possible to evaluate the diversity of fungi using a unified experimental method, even with samples obtained for diversity studies on other taxa such as those of animals.

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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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Environmental DNA metabarcoding for benthic monitoring: A review of sediment sampling and DNA extraction methods

The Science of The Total Environment ◽

10.1016/j.scitotenv.2021.151783 ◽

2021 ◽

pp. 151783

Author(s):

J. Pawlowski ◽

K. Bruce ◽

K. Panksep ◽

F.I. Aguirre ◽

S. Amafitano ◽

...

Keyword(s):

Dna Extraction ◽

Environmental Dna ◽

Extraction Methods ◽

Dna Metabarcoding ◽

Sediment Sampling ◽

Dna Extraction Methods

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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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