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Plant hormone profiling is therefore essential to understand developmental processes and the adaptation of plants to biotic and\/or abiotic stresses. Interestingly, commonly used hormone extraction and profiling methods do not adequately resolve other molecular entities, such as polar metabolites, lipids, starch and proteins, which would be required to comprehensively describe the continuing biological processes at a systematic level. In this article we introduce an updated version of a previously published liquid:liquid metabolite extraction protocol, which not only allows for the profiling of primary and secondary metabolites, lipids, starch and proteins, but also enables the quantitative analysis of the major plant hormone classes, including abscisic acid, auxins, cytokinins, jasmonates and salicylates, from a single sample aliquot. The optimization of the method, which uses the introduction of acidified water, enabling the complete purification of major plant hormones into the organic (methyl\u2010\n                    <jats:italic>tert<\/jats:italic>\n                    \u2010butyl\u2010ether) phase, eliminated the need for solid\u2010phase extraction for sample clean\u2010up, and therefore reduces both sampling time and cost. As a proof\u2010of\u2010concept analysis,\n                    <jats:italic>Arabidopsis thaliana<\/jats:italic>\n                    plants were subjected to water\u2010deficit stress, which were then profiled for hormonal, metabolic, lipidomic and proteomic changes. Surprisingly, we determined not only previously described molecular changes but also significant changes regarding the breakdown of specific galactolipids, followed by the substantial accumulation of unsaturated fatty\u2010acid derivatives and diverse jasmonates in the course of adaptation to water\u2010deficit stress.\n                  <\/jats:p>","DOI":"10.1111\/tpj.14800","type":"journal-article","created":{"date-parts":[[2020,5,7]],"date-time":"2020-05-07T06:14:12Z","timestamp":1588832052000},"page":"1614-1632","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":111,"title":["An improved extraction method enables the comprehensive analysis of lipids, proteins, metabolites and phytohormones from a single sample of leaf tissue under water\u2010deficit stress"],"prefix":"10.1111","volume":"103","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0332-3860","authenticated-orcid":false,"given":"Mohamed A.","family":"Salem","sequence":"first","affiliation":[{"name":"Department of Pharmacognosy Faculty of Pharmacy Menoufia University Gamal Abd El Nasr St Shibin Elkom Menoufia 32511 Egypt"},{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takuya","family":"Yoshida","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leonardo","family":"Perez de Souza","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2067-5235","authenticated-orcid":false,"given":"Saleh","family":"Alseekh","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"},{"name":"Center of Plant Systems Biology and Biotechnology Plovdiv 4000 Bulgaria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krzysztof","family":"Bajdzienko","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9000-335X","authenticated-orcid":false,"given":"Alisdair R.","family":"Fernie","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Giavalisco","sequence":"additional","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology Am M\u00fchlenberg 1 Potsdam\u2010Golm 14476 Germany"},{"name":"Max Planck Institute for Biology of Ageing Joseph Stelzmann Str. 9b Cologne 50931 Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,6,5]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1199\/tab.0129"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11103-016-0501-8"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00037021"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj2350025"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2016.02070"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-313X.2003.01800.x"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.92.10.4114"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2105-10-428"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2016.11.007"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2012.12.005"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2011.03.013"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1360-1385(01)02216-6"},{"key":"e_1_2_9_14_1","unstructured":"EMEA(2006)Note for Guidance on Validation of Analytical Procedures: Text and Methodology.European Medicines Agency. 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