In Figs 1 and 2 and in Table 2, the viability of cells determined as CFU is shown. The internal

K+ content in cells from the stationary growth phase was estimated as described earlier (Kinclova, et al., 2001). Briefly, cells (three aliquots per strain) were collected on Millipore membrane filters (0.8 μm pore diameter) and quickly washed with 20 mM MgCl2. The cells were then extracted with HCl and analyzed with a flame atomic absorption spectrophotometer. The experiments were repeated selleck chemicals llc three times. To characterize the role of plasma membrane potassium transporters upon cell dehydration and subsequent rehydration, we first estimated the desiccation survival of cells lacking either the two main potassium uptake systems (BYT12, trk1Δ trk2Δ), the two active potassium efflux systems (BYT45, nha1Δ ena1-5Δ) or all three K+ exporters (BYT345, tok1Δ nha1Δ ena1-5Δ). The experimental conditions (cf. ‘Materials and methods’) were set to

achieve c. 70% survival of the parental BY4741 strain, so that a better or worse survival rate of the mutants could be easily observed. All strains were grown in YPD supplemented with 50 mM KCl [to achieve a comparable growth of strains lacking the Trk transporters;(Navarette et al., 2010)] to the stationary phase of growth, as it has been repeatedly shown that exponentially growing cells are, compared with stationary cells, much more sensitive to various types of stress, including anhydrobiotic stress (Beker & Rapoport, 1987). Figure 1a shows that the absence of potassium exporting systems (BYT45 and BYT345 cells) did not significantly change the ability of cells to survive

find more dehydration/rehydration Oxymatrine treatment. About 65–70% of cells lacking potassium exporters were able to survive the desiccation and revitalization processes. On the other hand, the absence of potassium uptake systems (BYT12, trk1Δ trk2Δ) brought about a dramatic decrease in the survival rate. Only about 8% of cells were able to form colonies after dehydration/rehydration treatment. This result suggested the importance of potassium uptake for anhydrobiosis. To distinguish which of the two Trk transporters’ absence causes the observed phenotype, the same experiment was repeated with single mutants lacking either the Trk1 (BYT1) or Trk2 (BYT2) transporter. It was the absence of Trk2 that diminished the ability of cells to survive desiccation stress (Fig. 1b). Since the deletion of the TRK2 gene has almost no phenotype in exponential cells harboring an intact copy of TRK1 (Petrezselyova et al., 2011), we were aware of a risk of a non-specific mutation that could occur during the construction of the BYT2 mutant, e.g. upon electroporation. To be sure that the observed phenotype is related to the absence of the TRK2 gene and not to an additional non-specific mutation, we tested the survival of two independently prepared BYT1 (trk1Δ) and three BYT2 (trk2Δ) mutants (Fig. 2).