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However, how aquaporins are involved in the synergistic transport of Mg<jats:sup>2+<\/jats:sup> and water remains poorly understood. Here, we found that the cassava aquaporin MePIP2;7 was involved in Mg<jats:sup>2+<\/jats:sup> transport through interaction with MeMGT9, a lower affinity magnesium transporter protein. Knockdown of <jats:italic>MePIP2;7<\/jats:italic> in cassava led to magnesium deficiency in basal mature leaves with chlorosis and necrotic spots on their edges and starch over\u2010accumulation. Mg<jats:sup>2+<\/jats:sup> content was significantly decreased in leaves and roots of <jats:italic>MePIP2;7<\/jats:italic>\u2010RNA interference (PIP\u2010Ri) plants grown in both field and Mg<jats:sup>2+<\/jats:sup>\u2010free hydroponic solution. <jats:italic>Xenopus<\/jats:italic> oocyte injection analysis verified that MePIP2;7 possessed the ability to transport water only and MeMGT9 was responsible for Mg<jats:sup>2+<\/jats:sup> efflux. More importantly, MePIP2;7 improved the transportability of Mg<jats:sup>2+<\/jats:sup> via MeMGT9 as verified using the CM66 mutant complementation assay and <jats:italic>Xenopus<\/jats:italic> oocytes expressing system. Yeast two\u2010hybrid, bimolecular fluorescence complementation, co\u2010localization, and co\u2010immunoprecipitation assays demonstrated the direct protein\u2013protein interaction between MePIP2;7 and MeMGT9 <jats:italic>in vivo<\/jats:italic>. Mg<jats:sup>2+<\/jats:sup> flux was significantly elevated in <jats:italic>MePIP2;7<\/jats:italic>\u2010overexpressing lines in hydroponic solution through non\u2010invasive micro\u2010test technique analysis. Under Mg<jats:sup>2+<\/jats:sup>\u2010free condition, the retarded growth of PIP\u2010Ri transgenic plants could be recovered with Mg<jats:sup>2+<\/jats:sup> supplementation. Taken together, our results demonstrated the synergistic effect of the MePIP2;7 and MeMGT9 interaction in regulating water and Mg<jats:sup>2+<\/jats:sup> absorption and transport in cassava.<\/jats:p>","DOI":"10.1111\/jipb.13552","type":"journal-article","created":{"date-parts":[[2023,8,7]],"date-time":"2023-08-07T08:52:44Z","timestamp":1691398364000},"page":"2349-2367","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["The aquaporin MePIP2;7 improves MeMGT9\u2010mediated Mg<sup>2<\/sup><sup>+<\/sup> acquisition in cassava"],"prefix":"10.1111","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4689-5682","authenticated-orcid":false,"given":"Qiuxiang","family":"Ma","sequence":"first","affiliation":[{"name":"National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai 200032 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