Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field
Abstract Premise The gametophytes of different fern species collected in the field can be difficult to distinguish because of their morphological similarities. Nonetheless, emerging molecular ecology techniques are starting to be used to tackle such limitations. Here, using case studies and a detail...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-03-01
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| Series: | Applications in Plant Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/aps3.11462 |
| _version_ | 1818274902667427840 |
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| author | Yi‐Hsuan Wu Ya‐Ting Ke Yuan‐Yao Chan Goang‐Jiun Wang Li‐Yaung Kuo |
| author_facet | Yi‐Hsuan Wu Ya‐Ting Ke Yuan‐Yao Chan Goang‐Jiun Wang Li‐Yaung Kuo |
| author_sort | Yi‐Hsuan Wu |
| collection | DOAJ |
| description | Abstract Premise The gametophytes of different fern species collected in the field can be difficult to distinguish because of their morphological similarities. Nonetheless, emerging molecular ecology techniques are starting to be used to tackle such limitations. Here, using case studies and a detailed protocol, we demonstrate a convenient methodology, tissue‐direct PCR (TD‐PCR), that foregoes a traditional DNA extraction and facilitates the identification of fern gametophytes, as well as enabling the elucidation of their natural distribution. Methods Based on updated plastome information, we designed a universal primer set targeting the trnL‐L‐F region, which is effective across extant ferns. We used this primer set to perform TD‐PCR on the case‐studied populations of Taiwanese Lomariopsis gametophytes, using the generated sequences for their identification. In the case study concerning the microhabitat preference of Vaginularia junghuhnii, we designed and used a taxon‐specific primer set. Results Compared with approaches requiring DNA extraction, the use of TD‐PCR with either universal or taxon‐specific primers could save significant time, money, labor, and research materials in the genetic identification of fern gametophytes. Discussion The use of modern genetic tools can aid in the identification of fern gametophytes. An updated TD‐PCR strategy not only facilitates the DNA‐based identification of gametophytes, but also promotes new avenues of research for investigating these plants in the field. |
| first_indexed | 2024-12-12T22:21:14Z |
| format | Article |
| id | doaj.art-3de7fe2509904e34aa0ab7c331114930 |
| institution | Directory Open Access Journal |
| issn | 2168-0450 |
| language | English |
| last_indexed | 2024-12-12T22:21:14Z |
| publishDate | 2022-03-01 |
| publisher | Wiley |
| record_format | Article |
| series | Applications in Plant Sciences |
| spelling | doaj.art-3de7fe2509904e34aa0ab7c3311149302022-12-22T00:09:54ZengWileyApplications in Plant Sciences2168-04502022-03-01102n/an/a10.1002/aps3.11462Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the fieldYi‐Hsuan Wu0Ya‐Ting Ke1Yuan‐Yao Chan2Goang‐Jiun Wang3Li‐Yaung Kuo4Institute of Molecular and Cellular Biology National Tsing Hua University Hsinchu City TaiwanInstitute of Molecular and Cellular Biology National Tsing Hua University Hsinchu City TaiwanCollege of Biological Science and Technology National Yang Ming Chiao Tung University Hsinchu City TaiwanDepartment of Life Science, College of Life Science National Taiwan University Taipei City TaiwanInstitute of Molecular and Cellular Biology National Tsing Hua University Hsinchu City TaiwanAbstract Premise The gametophytes of different fern species collected in the field can be difficult to distinguish because of their morphological similarities. Nonetheless, emerging molecular ecology techniques are starting to be used to tackle such limitations. Here, using case studies and a detailed protocol, we demonstrate a convenient methodology, tissue‐direct PCR (TD‐PCR), that foregoes a traditional DNA extraction and facilitates the identification of fern gametophytes, as well as enabling the elucidation of their natural distribution. Methods Based on updated plastome information, we designed a universal primer set targeting the trnL‐L‐F region, which is effective across extant ferns. We used this primer set to perform TD‐PCR on the case‐studied populations of Taiwanese Lomariopsis gametophytes, using the generated sequences for their identification. In the case study concerning the microhabitat preference of Vaginularia junghuhnii, we designed and used a taxon‐specific primer set. Results Compared with approaches requiring DNA extraction, the use of TD‐PCR with either universal or taxon‐specific primers could save significant time, money, labor, and research materials in the genetic identification of fern gametophytes. Discussion The use of modern genetic tools can aid in the identification of fern gametophytes. An updated TD‐PCR strategy not only facilitates the DNA‐based identification of gametophytes, but also promotes new avenues of research for investigating these plants in the field.https://doi.org/10.1002/aps3.11462DNA‐based identificationfern gametophyteindependent gametophytesLomariopsismolecular ecologytissue‐direct PCR |
| spellingShingle | Yi‐Hsuan Wu Ya‐Ting Ke Yuan‐Yao Chan Goang‐Jiun Wang Li‐Yaung Kuo Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field Applications in Plant Sciences DNA‐based identification fern gametophyte independent gametophytes Lomariopsis molecular ecology tissue‐direct PCR |
| title | Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field |
| title_full | Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field |
| title_fullStr | Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field |
| title_full_unstemmed | Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field |
| title_short | Integrating tissue‐direct PCR into genetic identification: An upgraded molecular ecology approach to survey fern gametophytes in the field |
| title_sort | integrating tissue direct pcr into genetic identification an upgraded molecular ecology approach to survey fern gametophytes in the field |
| topic | DNA‐based identification fern gametophyte independent gametophytes Lomariopsis molecular ecology tissue‐direct PCR |
| url | https://doi.org/10.1002/aps3.11462 |
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