{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,13]],"date-time":"2026-04-13T22:41:38Z","timestamp":1776120098187,"version":"3.50.1"},"reference-count":34,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,1,19]],"date-time":"2025-01-19T00:00:00Z","timestamp":1737244800000},"content-version":"vor","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["bsapubs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Appl Plant Sci"],"published-print":{"date-parts":[[2025,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Premise<\/jats:title><jats:p>Mechanistic models using stomatal traits and leaf carbon isotope ratios to reconstruct atmospheric carbon dioxide (CO<jats:sub>2<\/jats:sub>) concentrations (<jats:italic>c<\/jats:italic><jats:sub><jats:italic>a<\/jats:italic><\/jats:sub>) are important to understand the Phanerozoic paleoclimate. However, methods for preparing leaf cuticles to measure stomatal traits have not been standardized.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Three people measured the stomatal density and index, guard cell length, guard cell pair width, and pore length of leaves from the same <jats:italic>Ginkgo biloba<\/jats:italic>, <jats:italic>Quercus alba<\/jats:italic>, and <jats:italic>Zingiber mioga<\/jats:italic> leaves growing at known CO<jats:sub>2<\/jats:sub> levels using four preparation methods: fluorescence on cleared leaves, nail polish, dental putty on fresh leaves, and dental putty on dried leaves.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>There are significant differences between trait measurements from each method. Modeled <jats:italic>c<\/jats:italic><jats:sub><jats:italic>a<\/jats:italic><\/jats:sub> calculations are less sensitive to method than individual traits; however, the choice of assumed carbon isotope fractionation also impacted the accuracy of the results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Discussion<\/jats:title><jats:p>We show that there is not a significant difference between <jats:italic>c<\/jats:italic><jats:sub><jats:italic>a<\/jats:italic><\/jats:sub> estimates made using any of the four methods. Further study is needed on the fractionation due to carboxylation of ribulose bisphosphate (RuBP) in individual plant species before use as a paleo\u2010CO<jats:sub>2<\/jats:sub> barometer and to refine estimates based upon widely applied taxa (e.g., <jats:italic>Ginkgo<\/jats:italic>). Finally, we recommend that morphological measurements be made by multiple observers to reduce the effect of individual observational biases.<\/jats:p><\/jats:sec>","DOI":"10.1002\/aps3.11629","type":"journal-article","created":{"date-parts":[[2025,1,20]],"date-time":"2025-01-20T00:36:59Z","timestamp":1737333419000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["The sensitivity of reconstructed carbon dioxide concentrations to stomatal preparation methods using a leaf gas exchange model"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-3959-4248","authenticated-orcid":false,"given":"Michael D.","family":"Machesky","sequence":"first","affiliation":[{"name":"Department of Earth and Environmental Sciences University of Michigan 1100 N. 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