Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein
Premise Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whethe...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2020-05-01
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| Series: | Applications in Plant Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/aps3.11346 |
| _version_ | 1818468104406040576 |
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| author | Eastyn L. Newsome Grace L. Brock Jared Lutz Robert L. Baker |
| author_facet | Eastyn L. Newsome Grace L. Brock Jared Lutz Robert L. Baker |
| author_sort | Eastyn L. Newsome |
| collection | DOAJ |
| description | Premise Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of Brassica rapa. Methods FAA (formaldehyde, glacial acetic acid, and ethanol)‐fixed samples were stored in ethanol. Without performing any additional clearing or staining, we tested two methods of image acquisition at three locations along the proximal‐distal axis of the laminae and estimated the patterns of venation using the program phenoVein. We developed and made available an R script to handle the phenoVein output and then analyzed our data using linear mixed‐effects models. Results Beyond fixation and storage, staining and clearing are not necessary to estimate leaf venation using phenoVein if the images are acquired using a stereomicroscope. All estimates of venation required some manual adjustment. We found a significant effect of location within the laminae for all aspects of venation. Discussion By removing the clearing and staining steps and utilizing the semi‐automated program phenoVein, we quickly and cheaply acquired leaf venation data. Venation may be an important target for crop breeding efforts, particularly if intralaminar variation correlates with variation in physiological processes, which remains an open question. |
| first_indexed | 2024-04-13T21:09:23Z |
| format | Article |
| id | doaj.art-6519a91f032f4b908ef4585e8e06c87f |
| institution | Directory Open Access Journal |
| issn | 2168-0450 |
| language | English |
| last_indexed | 2024-04-13T21:09:23Z |
| publishDate | 2020-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Applications in Plant Sciences |
| spelling | doaj.art-6519a91f032f4b908ef4585e8e06c87f2022-12-22T02:29:52ZengWileyApplications in Plant Sciences2168-04502020-05-0185n/an/a10.1002/aps3.11346Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVeinEastyn L. Newsome0Grace L. Brock1Jared Lutz2Robert L. Baker3Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USADepartment of Biology Miami University 700 E High Street Oxford Ohio 45056 USADepartment of Biology Miami University 700 E High Street Oxford Ohio 45056 USADepartment of Biology Miami University 700 E High Street Oxford Ohio 45056 USAPremise Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of Brassica rapa. Methods FAA (formaldehyde, glacial acetic acid, and ethanol)‐fixed samples were stored in ethanol. Without performing any additional clearing or staining, we tested two methods of image acquisition at three locations along the proximal‐distal axis of the laminae and estimated the patterns of venation using the program phenoVein. We developed and made available an R script to handle the phenoVein output and then analyzed our data using linear mixed‐effects models. Results Beyond fixation and storage, staining and clearing are not necessary to estimate leaf venation using phenoVein if the images are acquired using a stereomicroscope. All estimates of venation required some manual adjustment. We found a significant effect of location within the laminae for all aspects of venation. Discussion By removing the clearing and staining steps and utilizing the semi‐automated program phenoVein, we quickly and cheaply acquired leaf venation data. Venation may be an important target for crop breeding efforts, particularly if intralaminar variation correlates with variation in physiological processes, which remains an open question.https://doi.org/10.1002/aps3.11346areole areaBrassica rapaleaf venationphenoVeinvenation density, venation variation |
| spellingShingle | Eastyn L. Newsome Grace L. Brock Jared Lutz Robert L. Baker Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein Applications in Plant Sciences areole area Brassica rapa leaf venation phenoVein venation density, venation variation |
| title | Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein |
| title_full | Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein |
| title_fullStr | Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein |
| title_full_unstemmed | Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein |
| title_short | Variation within laminae: Semi‐automated methods for quantifying leaf venation using phenoVein |
| title_sort | variation within laminae semi automated methods for quantifying leaf venation using phenovein |
| topic | areole area Brassica rapa leaf venation phenoVein venation density, venation variation |
| url | https://doi.org/10.1002/aps3.11346 |
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