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Effect of fuchsin fixation of pollen on DNA barcode recovery

Pollen grains attached to insects are a valuable source of ecological information which can be used to reconstruct visitation networks. Morphological pollen identification relies on light microscopy with pollen usually stained and mounted in fuchsin jelly, which is also used to remove pollen from th...

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Autores principales: Streicher, Melanie B., Johnson, Steven D., Willows‐Munro, Sandi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10468989/
https://www.ncbi.nlm.nih.gov/pubmed/37664513
http://dx.doi.org/10.1002/ece3.10475
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author Streicher, Melanie B.
Johnson, Steven D.
Willows‐Munro, Sandi
author_facet Streicher, Melanie B.
Johnson, Steven D.
Willows‐Munro, Sandi
author_sort Streicher, Melanie B.
collection PubMed
description Pollen grains attached to insects are a valuable source of ecological information which can be used to reconstruct visitation networks. Morphological pollen identification relies on light microscopy with pollen usually stained and mounted in fuchsin jelly, which is also used to remove pollen from the bodies of insects. Pollen embedded in fuchsin jelly could potentially be used for DNA barcoding and metabarcoding (large‐scale taxonomic identification of complex mixed samples) and thus provide additional information for pollination networks. In this study, we determine whether fuchsin‐embedded pollen can be used for downstream molecular applications. We evaluate the quality of plant barcode (ITS) sequences amplified from DNA extracted from both fresh (untreated) pollen, and pollen which had been embedded in fuchsin jelly. We show that the addition of fuchsin to DNA extraction does not impact DNA barcode sequence quality during short‐term storage. DNA extractions from both untreated and fuchsin‐treated pollen produced reliable barcode sequences of high quality. Our findings suggest that pollen which has been collected, stained, and embedded in fuchsin jelly for preliminary microscopy work can be used within several days for downstream genetic analysis, though the quality of DNA from pollen stored in fuchsin jelly for extended periods is yet to be established.
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spelling pubmed-104689892023-09-01 Effect of fuchsin fixation of pollen on DNA barcode recovery Streicher, Melanie B. Johnson, Steven D. Willows‐Munro, Sandi Ecol Evol Research Articles Pollen grains attached to insects are a valuable source of ecological information which can be used to reconstruct visitation networks. Morphological pollen identification relies on light microscopy with pollen usually stained and mounted in fuchsin jelly, which is also used to remove pollen from the bodies of insects. Pollen embedded in fuchsin jelly could potentially be used for DNA barcoding and metabarcoding (large‐scale taxonomic identification of complex mixed samples) and thus provide additional information for pollination networks. In this study, we determine whether fuchsin‐embedded pollen can be used for downstream molecular applications. We evaluate the quality of plant barcode (ITS) sequences amplified from DNA extracted from both fresh (untreated) pollen, and pollen which had been embedded in fuchsin jelly. We show that the addition of fuchsin to DNA extraction does not impact DNA barcode sequence quality during short‐term storage. DNA extractions from both untreated and fuchsin‐treated pollen produced reliable barcode sequences of high quality. Our findings suggest that pollen which has been collected, stained, and embedded in fuchsin jelly for preliminary microscopy work can be used within several days for downstream genetic analysis, though the quality of DNA from pollen stored in fuchsin jelly for extended periods is yet to be established. John Wiley and Sons Inc. 2023-08-31 /pmc/articles/PMC10468989/ /pubmed/37664513 http://dx.doi.org/10.1002/ece3.10475 Text en © 2023 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Streicher, Melanie B.
Johnson, Steven D.
Willows‐Munro, Sandi
Effect of fuchsin fixation of pollen on DNA barcode recovery
title Effect of fuchsin fixation of pollen on DNA barcode recovery
title_full Effect of fuchsin fixation of pollen on DNA barcode recovery
title_fullStr Effect of fuchsin fixation of pollen on DNA barcode recovery
title_full_unstemmed Effect of fuchsin fixation of pollen on DNA barcode recovery
title_short Effect of fuchsin fixation of pollen on DNA barcode recovery
title_sort effect of fuchsin fixation of pollen on dna barcode recovery
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10468989/
https://www.ncbi.nlm.nih.gov/pubmed/37664513
http://dx.doi.org/10.1002/ece3.10475
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