Fig 1.
Subcellular localization studies of VvXIP1-RFP in tobacco epidermal cells (A-P).
VvXIP1-RFP was co-transformed with YFP-ZmPIP2;5 (plasma membrane marker, A-D), YFP-TIP1;2 (tonoplast marker, E-H), ST-YFP (Golgi marker, I-L), and YFP-HDEL (ER marker, M-P).
Fig 2.
Study of VvXIP1 water and glycerol permeability in yeast.
Normalized scattered light intensity obtained from stopped flow experiments performed at 28°C with membrane vesicles collected from yeast cells transformed with pVV214-VvXIP1 (grey) or the empty vector (black), suddenly exposed to an osmotic gradient of 240 mOsM with a mannitol solution (A) and with a glycerol solution (B).
Table 1.
Permeability (Pf and Pgly) and activation energy (Ea) for water transport in yeast membranes obtained by stopped flow (for details see Material and Methods).
Fig 3.
H2O2 transport by yeast cells expressing VvXIP1.
Yeast cultures transformed with pVV214-VvXIP1 (black line) and control cells (empty vector, grey line) were incubated overnight with the CM-H2DCFDA probe and fluorescence increase after the addition of 0.6 mM H2O2 was monitored in a spectrofluorimeter (A). O2 release (after the intracellular breakdown of H2O2) by cells transformed with pVV214-VvXIP1 (black) and control yeast (grey) monitored with a Clark electrode in response to 50 μM H2O2 (solid lines) and after mercury inhibition (dashed lines) (B). Plate growth assays in YNB media supplemented with 0.6 mM H2O2 (C).
Fig 4.
The effect of metals and metalloids in the growth of S. cerevisiae overexpressing VvXIP1.
Yeast transformed with pVV214-VvXIP1 and control cells (empty vector) were diluted (OD600 = 0.1, 0.01 and 0.001) and spotted on medium containing 0.45 mM H3AsO4, 40 mM H3BO3, 0.5 mM NiCl2 and 5 mM CuSO4 and the growth was recorded following 3–5 days at 30°C.
Fig 5.
Study of VvXIP1 expression in berries, canes, flowers and leaves from grapevine cv.
Vinhão (A) grown under field conditions. VvXIP1 steady-state transcript levels in leaves from potted grapevines (cv. Aragonez) under drought stress (B) and sorbitol induced osmotic stress sensitivity of yeast cells expressing VvXIP1 (C).
Fig 6.
VvXIP1 expression in grape cultured cells in response to hormones, salt, copper and boron.
CSB cells were incubated overnight with SA (150 μM), ABA (150 μM), NaCl (100 mM), copper (100 μM) and boron (100 μM), and the expression of VvXIP1 was studied by qRT-PCR.