Figure 3. Localization and Pi transport activity of BnPHT5;1b proteins.
(a, b) 3-D structure comparison of BnPHT5;1b proteins and AtVPT1. The Alphafold2 tool was used to predict protein 3-D structure. Pymol was used to calculate the similarity. (c) Tonoplast localization of BnA09PHT5;1b and BnCnPHT5;1b. GFP-labeled BnPHT5;1b constructs were expressed in mesophyll protoplast from leaves of 3-4-week old Arabidopsis seedlings and the GFP signal was recorded using a Leica SP8 confocal microscope. Scale bar = 5 μm. All experiments were repeated twice, and similar results were obtained.(d) BnPHT5;1b confers Pi transport activity in yeast. Constructs including BnPHT5;1b were expressed individually in YP100 (Δpho84Δpho87Δpho89Δpho90Δpho91Δgit1). Strains were diluted to OD600 values of 1, 0.1, 0.01 and dripped on YNB medium (pH 5.6) supplemented with glucose and 30 mM Pi for 5 days incubation at 30 °C. All experiments were repeated three times, and similar results were obtained.