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A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae
PREMISE: Putatively single‐copy nuclear (SCN) loci, which are identified using genomic resources of closely related species, are ideal for phylogenomic inference. However, suitable genomic resources are not available for many clades, including Melastomataceae. We introduce a versatile approach to id...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7249273/ https://www.ncbi.nlm.nih.gov/pubmed/32477841 http://dx.doi.org/10.1002/aps3.11345 |
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author | Jantzen, Johanna R. Amarasinghe, Prabha Folk, Ryan A. Reginato, Marcelo Michelangeli, Fabian A. Soltis, Douglas E. Cellinese, Nico Soltis, Pamela S. |
author_facet | Jantzen, Johanna R. Amarasinghe, Prabha Folk, Ryan A. Reginato, Marcelo Michelangeli, Fabian A. Soltis, Douglas E. Cellinese, Nico Soltis, Pamela S. |
author_sort | Jantzen, Johanna R. |
collection | PubMed |
description | PREMISE: Putatively single‐copy nuclear (SCN) loci, which are identified using genomic resources of closely related species, are ideal for phylogenomic inference. However, suitable genomic resources are not available for many clades, including Melastomataceae. We introduce a versatile approach to identify SCN loci for clades with few genomic resources and use it to develop probes for target enrichment in the distantly related Memecylon and Tibouchina (Melastomataceae). METHODS: We present a two‐tiered pipeline. First, we identified putatively SCN loci using MarkerMiner and transcriptomes from distantly related species in Melastomataceae. Published loci and genes of functional significance were then added (384 total loci). Second, using HybPiper, we retrieved 689 homologous template sequences for these loci using genome‐skimming data from within the focal clades. RESULTS: We sequenced 193 loci common to Memecylon and Tibouchina. Probes designed from 56 template sequences successfully targeted sequences in both clades. Probes designed from genome‐skimming data within a focal clade were more successful than probes designed from other sources. DISCUSSION: Our pipeline successfully identified and targeted SCN loci in Memecylon and Tibouchina, enabling phylogenomic studies in both clades and potentially across Melastomataceae. This pipeline could be easily applied to other clades with few genomic resources. |
format | Online Article Text |
id | pubmed-7249273 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-72492732020-05-29 A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae Jantzen, Johanna R. Amarasinghe, Prabha Folk, Ryan A. Reginato, Marcelo Michelangeli, Fabian A. Soltis, Douglas E. Cellinese, Nico Soltis, Pamela S. Appl Plant Sci Application Articles PREMISE: Putatively single‐copy nuclear (SCN) loci, which are identified using genomic resources of closely related species, are ideal for phylogenomic inference. However, suitable genomic resources are not available for many clades, including Melastomataceae. We introduce a versatile approach to identify SCN loci for clades with few genomic resources and use it to develop probes for target enrichment in the distantly related Memecylon and Tibouchina (Melastomataceae). METHODS: We present a two‐tiered pipeline. First, we identified putatively SCN loci using MarkerMiner and transcriptomes from distantly related species in Melastomataceae. Published loci and genes of functional significance were then added (384 total loci). Second, using HybPiper, we retrieved 689 homologous template sequences for these loci using genome‐skimming data from within the focal clades. RESULTS: We sequenced 193 loci common to Memecylon and Tibouchina. Probes designed from 56 template sequences successfully targeted sequences in both clades. Probes designed from genome‐skimming data within a focal clade were more successful than probes designed from other sources. DISCUSSION: Our pipeline successfully identified and targeted SCN loci in Memecylon and Tibouchina, enabling phylogenomic studies in both clades and potentially across Melastomataceae. This pipeline could be easily applied to other clades with few genomic resources. John Wiley and Sons Inc. 2020-05-09 /pmc/articles/PMC7249273/ /pubmed/32477841 http://dx.doi.org/10.1002/aps3.11345 Text en © 2020 Jantzen et al. Applications in Plant Sciences is published by Wiley Periodicals, Inc. on behalf of the Botanical Society of America This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Application Articles Jantzen, Johanna R. Amarasinghe, Prabha Folk, Ryan A. Reginato, Marcelo Michelangeli, Fabian A. Soltis, Douglas E. Cellinese, Nico Soltis, Pamela S. A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title | A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title_full | A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title_fullStr | A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title_full_unstemmed | A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title_short | A two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in Melastomataceae |
title_sort | two‐tier bioinformatic pipeline to develop probes for target capture of nuclear loci with applications in melastomataceae |
topic | Application Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7249273/ https://www.ncbi.nlm.nih.gov/pubmed/32477841 http://dx.doi.org/10.1002/aps3.11345 |
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