The aquaporin MePIP2;7 improves MeMGT9-mediated Mg 2+ acquisition in cassava.

Aquaporins are important transmembrane water transport proteins that transport water and several neutral molecules. However, how aquaporins are involved in the synergistic transport of Mg 2+ and water remains poorly understood. Here, we found that the cassava aquaporin MePIP2;7 was involved in Mg 2+ transport through interaction with MeMGT9, a lower affinity magnesium transporter protein. Knockdown of MePIP2;7 in cassava led to magnesium deficiency in basal mature leaves with chlorosis and necrotic spots on their edges and starch over-accumulation. Mg 2+ content was significantly decreased in leaves and roots of MePIP2;7-RNAi plants grown in both field and hydroponic solution with Mg 2+ -free. Xenopus oocyte injection analysis verified that MePIP2;7 possessed the ability to transport water only and MeMGT9 was responsible for Mg 2+ efflux. More importantly, MePIP2;7 improved the transportability of Mg 2+ via MeMGT9 as verified using the CM66 mutant complementation assay and Xenopus oocytes expressing system. Yeast-two-hybrid, BiFC, co-localization and Co-IP assays demonstrated the direct protein-protein interaction between MePIP2;7 and MeMGT9 in vivo. Mg 2+ flux was significantly elevated in MePIP2;7-overexpressing lines in hydroponic solution through Non-invasive Micro-test Technique analysis. Under Mg 2+ -free condition, the retarded growth of MePIP2;7-RNAi transgenic plants could be recovered with Mg 2+ supplementation. Taken together, our results demonstrated the synergistic effect of the MePIP2;7 and MeMGT9 interaction in regulating water and Mg 2+ absorption and transport in cassava. This article is protected by copyright. All rights reserved.