{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T10:06:02Z","timestamp":1767261962042,"version":"build-2065373602"},"reference-count":22,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2020,5,11]],"date-time":"2020-05-11T00:00:00Z","timestamp":1589155200000},"content-version":"am","delay-in-days":10,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2020,5,11]],"date-time":"2020-05-11T00:00:00Z","timestamp":1589155200000},"content-version":"vor","delay-in-days":10,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["IOS\u20101306574"],"award-info":[{"award-number":["IOS\u20101306574"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006686","name":"University of Miami","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100006686","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["bsapubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Appl Plant Sci"],"published-print":{"date-parts":[[2020,5]]},"abstract":"<jats:sec><jats:title>Premise<\/jats:title><jats:p>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 <jats:italic>Brassica rapa<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">FAA<\/jats:styled-content> (formaldehyde, glacial acetic acid, and ethanol)\u2010fixed 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\u2010distal 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\u2010effects models.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>By removing the clearing and staining steps and utilizing the semi\u2010automated 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.<\/jats:p><\/jats:sec>","DOI":"10.1002\/aps3.11346","type":"journal-article","created":{"date-parts":[[2020,5,11]],"date-time":"2020-05-11T21:59:54Z","timestamp":1589234394000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Variation within laminae: Semi\u2010automated methods for quantifying leaf venation using phenoVein"],"prefix":"10.1002","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9975-7083","authenticated-orcid":false,"given":"Eastyn L.","family":"Newsome","sequence":"first","affiliation":[{"name":"Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6050-2577","authenticated-orcid":false,"given":"Grace L.","family":"Brock","sequence":"additional","affiliation":[{"name":"Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jared","family":"Lutz","sequence":"additional","affiliation":[{"name":"Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7591-5035","authenticated-orcid":false,"given":"Robert L.","family":"Baker","sequence":"additional","affiliation":[{"name":"Department of Biology Miami University 700 E High Street Oxford Ohio 45056 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,5,11]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12870-016-0957-3"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1534\/g3.117.300350"},{"key":"e_1_2_8_4_1","first-page":"2359","article-title":"phenoVein\u2014A tool for leaf vein segmentation and analysis","volume":"169","author":"B\u00fchler J.","year":"2015","journal-title":"Plant Physiology"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1002\/9781118448908.ch19"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00227-001-0760-y"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0094947"},{"key":"e_1_2_8_8_1","first-page":"102","article-title":"Ethnobotany and evolutionary origin of Indian oleiferous Brassicae","volume":"44","author":"Kokichi H.","year":"1984","journal-title":"Indian Journal of Genetics and Plant Breeding"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.127613"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.18637\/jss.v082.i13"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1360\/972013-721"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0066016"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.134072"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3040.2008.01798.x"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3040.2011.02470.x"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1111\/mec.14131"},{"volume-title":"R: A language and environment for statistical computing","year":"2019","author":"R Core Team","key":"e_1_2_8_17_1"},{"volume-title":"Biology of plants","year":"2005","author":"Raven P. 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