Research Paper Volume 14, Issue 7 pp 2989—3029

The tobacco phosphatidylethanolamine-binding protein NtFT4 increases the lifespan of Drosophila melanogaster by interacting with the proteostasis network


Figure 1. Expression of animal PEBPs in tobacco and Arabidopsis. (A) Bimolecular fluorescence complementation (BiFC) in infiltrated Nicotiana benthamiana leaves, representatively showing the interaction between Drosophila PEBP (NmRFP-CG7054) and NtFD1 (CmRFP-NtFD1). (B) BiFC representatively showing the interaction between Drosophila PEBP (NmRFP-CG7054) and tobacco 14-3-3 c (CmRFP-14-3-3 c). Scale bar = 50 μm. (C) Flowering time of tobacco lines expressing PEBP1, CG7054, CG7054-DS, RKIP or hPEBP4 under the control of the cauliflower mosaic virus 35S promoter. Abbreviation: VC: vector control. Flowering time was measured under long-day (LD) conditions in days after potting (dap). Data are means ± SEM, n = 50 (PEBP1, CG7054, CG7054-DS, RKIP and hPEBP4), n = 10 (VC). Significance was tested by one-way ANOVA and Tukey’s post hoc test (b significant compared with PEBP1, c significant compared with CG7054, all other comparisons non-significant). (D) Representative image of a transgenic tobacco plant expressing RKIP compared with the VC. Flowering time (E) and rosette leaf number at the onset of flowering (F) of transgenic Arabidopsis lines expressing RKIP, hPEBP4, CG7054, PEBP1 or the floral inducer NtFT4 under the control of the quadruple cauliflower 35S promoter. Col-0 = wild type A. thaliana Col-0 ecotype used for transformation. Flowering time was measured under LD conditions in days after seeding (das). Data are means ± SEM, n = 30 (CG7054, CG19594), n = 29 (hPEBP4), n = 19 (RKIP), n = 10 (Col-0), n = 8 (NtFT4); ****p < 0.001 in all pairwise comparisons with NtFT4 (a significant compared with Col-0 (p = 0.091) with all other comparisons being non-significant). Abbreviation: NS: no significant differences in any pairwise comparison. All p-values are provided in Supplementary Table 9. (G) Representative images of transgenic Arabidopsis plants expressing different PEBPs. Col-0 wild type plants (far right), and early flowering Q35-S:NtFT4 (left) and late flowering Q35-S:NtFT2 (far left) plants are shown in comparison with plants expressing the animal PEBPs.