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Modulating stomatal density can alter this exchange and thus presents a viable target for engineering improved crop productivity and climate resilience. Here, we show that stomatal density in <jats:italic>Arabidopsis thaliana<\/jats:italic> can be decreased by the expression of a water\u2010forming NAD(P)H oxidase targeted to stomatal precursor cells. We demonstrate that this reduction in stomatal density occurs irrespective of whether the expressed enzyme is localized to the cytosol, chloroplast stroma or chloroplast intermembrane space of these cells. We also reveal that this decrease in stomatal density occurs in the absence of any measurable impact on the efficiency and thermal sensitivity of photosynthesis,\u00a0or on stomatal dynamics. Consequently, overexpression plants exhibit a higher intrinsic water\u2010use efficiency due to an increase in CO<jats:sub>2<\/jats:sub> fixed per unit water transpired. Finally, we demonstrate that this enhanced water\u2010use efficiency translates to an improvement in vegetative growth and biomass accumulation under water\u2010deficit conditions. Together, these results thus provide a novel approach for enhancing plant productivity through metabolic engineering of stomatal density.<\/jats:p>","DOI":"10.1111\/pbi.70130","type":"journal-article","created":{"date-parts":[[2025,5,23]],"date-time":"2025-05-23T16:20:52Z","timestamp":1748017252000},"page":"3177-3194","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Metabolic engineering of stomatal precursor cells enhances photosynthetic water\u2010use efficiency and vegetative growth under water\u2010deficit conditions in <i>Arabidopsis thaliana<\/i>"],"prefix":"10.1111","volume":"23","author":[{"given":"Jacques W.","family":"Bouvier","sequence":"first","affiliation":[{"name":"Department of Biology University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8583-5362","authenticated-orcid":false,"given":"Steven","family":"Kelly","sequence":"additional","affiliation":[{"name":"Department of Biology University of Oxford  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,5,23]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/jxb\/erac218"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2015.01.025"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-3040.2011.02448.x"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.envexpbot.2016.12.004"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1111\/nph.16341"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0304-4238(91)90128-L"},{"key":"e_1_2_10_8_1","first-page":"723","article-title":"Epidermal pavement cells of Arabidopsis have chloroplasts","volume":"171","author":"Barton K.A.","year":"2016","journal-title":"Plant Physiol."},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.202275"},{"key":"e_1_2_10_10_1","first-page":"1603","article-title":"Stomatal responses of the \u2018living fossil\u2019 Ginkgo biloba L. to changes in atmospheric CO2 concentrations","volume":"49","author":"Beerling D.J.","year":"1998","journal-title":"J. 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