Effects of expressing GmCLC1 on the survival of the Δgef1 yeast mutant under MnCl2, NaCl, or KCl. (A)
GmCLC1 could enhance the survival of the chloride channel-deficient Δgef1 yeast mutant in the same way as GEF1, when the yeast was grown in media supplemented with MnCl2, KCl, and NaCl. The Δgef1 mutant was also transformed with an empty vector as a negative control. The expression vector pYES2 containing GEF1 or GmCLC1 was introduced into the Δgef1 mutant. Ten-fold serial dilutions (starting at OD550 ≈ 0.5) of each sample were plated on media of YPD, YPG, or YPG supplemented with 3 mM MnCl2, 1.0 M KCl, or 1.0 M NaCl. Plates were incubated at 30°C and photographs were taken after 60 h. (B) The expression of GmCLC1 in the gef1 yeast mutant increased Cl- accumulation. Cells of the above-mentioned Δgef1 mutant, Δgef1/Vector, Δgef1/GEF1, or Δgef1/GmCLC1 were grown in liquid YPG medium with 1.0 M NaCl. When cultures reached OD550 ≈ 0.2, cells were collected by filtration, and their Cl- contents were determined. DW, dry weigh. Data shown are the means and standard deviations of ion contents of three independent cultures of each strain. ∗∗: The difference between was significant at p < 0.01, using Duncan’s test.