Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA
Abstract Environmental DNA (eDNA) once shed can exist in numerous states with varying behaviors including degradation rates and transport potential. In this study, we consider three states of eDNA: (1) a membrane‐bound state referring to DNA enveloped in a cellular or organellar membrane, (2) a diss...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-05-01
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| Series: | Environmental DNA |
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| Online Access: | https://doi.org/10.1002/edn3.417 |
| _version_ | 1797823294855970816 |
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| author | Anish Kirtane Hannah Kleyer Kristy Deiner |
| author_facet | Anish Kirtane Hannah Kleyer Kristy Deiner |
| author_sort | Anish Kirtane |
| collection | DOAJ |
| description | Abstract Environmental DNA (eDNA) once shed can exist in numerous states with varying behaviors including degradation rates and transport potential. In this study, we consider three states of eDNA: (1) a membrane‐bound state referring to DNA enveloped in a cellular or organellar membrane, (2) a dissolved state defined as the extracellular DNA molecule in the environment without any interaction with other particles, and (3) an adsorbed state defined as extracellular DNA adsorbed to a particle surface in the environment. Capturing, isolating, and analyzing a target state of eDNA provides utility for better interpretation of eDNA degradation rates and transport potential. While methods for separating different states of DNA have been developed, they remain poorly evaluated due to the lack of state‐controlled experimentation. We evaluated the methods for separating states of eDNA from a single sample by spiking DNA from three different species to represent the three states of eDNA as state‐specific controls. We used chicken DNA to represent the dissolved state, cultured mouse cells for the membrane‐bound state, and salmon DNA adsorbed to clay particles as the adsorbed state. We performed the separation in three water matrices, two environmental and one synthetic, spiked with the three eDNA states. The membrane‐bound state was the only state that was isolated with minimal contamination from nontarget states. The membrane‐bound state also had the highest recovery (54.11 ± 19.24%), followed by the adsorbed state (5.08 ± 2.28%), and the dissolved state had the lowest total recovery (2.21 ± 2.36%). This study highlights the potential to sort the states of eDNA from a single sample and independently analyze them for more informed biodiversity assessments. However, further method development is needed to improve recovery and reduce cross‐contamination. |
| first_indexed | 2024-03-13T10:20:52Z |
| format | Article |
| id | doaj.art-919b4c6be8fc419d8e0493a44b7bc507 |
| institution | Directory Open Access Journal |
| issn | 2637-4943 |
| language | English |
| last_indexed | 2024-03-13T10:20:52Z |
| publishDate | 2023-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Environmental DNA |
| spelling | doaj.art-919b4c6be8fc419d8e0493a44b7bc5072023-05-20T12:29:57ZengWileyEnvironmental DNA2637-49432023-05-015358259610.1002/edn3.417Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNAAnish Kirtane0Hannah Kleyer1Kristy Deiner2Institute of Biogeochemistry and Pollutant Dynamics Eidgenössische Technische Hochschule Zürich Zürich SwitzerlandInstitute of Biogeochemistry and Pollutant Dynamics Eidgenössische Technische Hochschule Zürich Zürich SwitzerlandInstitute of Biogeochemistry and Pollutant Dynamics Eidgenössische Technische Hochschule Zürich Zürich SwitzerlandAbstract Environmental DNA (eDNA) once shed can exist in numerous states with varying behaviors including degradation rates and transport potential. In this study, we consider three states of eDNA: (1) a membrane‐bound state referring to DNA enveloped in a cellular or organellar membrane, (2) a dissolved state defined as the extracellular DNA molecule in the environment without any interaction with other particles, and (3) an adsorbed state defined as extracellular DNA adsorbed to a particle surface in the environment. Capturing, isolating, and analyzing a target state of eDNA provides utility for better interpretation of eDNA degradation rates and transport potential. While methods for separating different states of DNA have been developed, they remain poorly evaluated due to the lack of state‐controlled experimentation. We evaluated the methods for separating states of eDNA from a single sample by spiking DNA from three different species to represent the three states of eDNA as state‐specific controls. We used chicken DNA to represent the dissolved state, cultured mouse cells for the membrane‐bound state, and salmon DNA adsorbed to clay particles as the adsorbed state. We performed the separation in three water matrices, two environmental and one synthetic, spiked with the three eDNA states. The membrane‐bound state was the only state that was isolated with minimal contamination from nontarget states. The membrane‐bound state also had the highest recovery (54.11 ± 19.24%), followed by the adsorbed state (5.08 ± 2.28%), and the dissolved state had the lowest total recovery (2.21 ± 2.36%). This study highlights the potential to sort the states of eDNA from a single sample and independently analyze them for more informed biodiversity assessments. However, further method development is needed to improve recovery and reduce cross‐contamination.https://doi.org/10.1002/edn3.417adsorbed DNAdissolved DNAeDNAmembrane‐bound DNAqPCRstate sorting |
| spellingShingle | Anish Kirtane Hannah Kleyer Kristy Deiner Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA Environmental DNA adsorbed DNA dissolved DNA eDNA membrane‐bound DNA qPCR state sorting |
| title | Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA |
| title_full | Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA |
| title_fullStr | Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA |
| title_full_unstemmed | Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA |
| title_short | Sorting states of environmental DNA: Effects of isolation method and water matrix on the recovery of membrane‐bound, dissolved, and adsorbed states of eDNA |
| title_sort | sorting states of environmental dna effects of isolation method and water matrix on the recovery of membrane bound dissolved and adsorbed states of edna |
| topic | adsorbed DNA dissolved DNA eDNA membrane‐bound DNA qPCR state sorting |
| url | https://doi.org/10.1002/edn3.417 |
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