Variation in specific conductivity and major ions in streams must be understood to assess the effects of changes in ionic strength and salinity on stream biota. I compiled data for randomly selected sites from surveys conducted from 1985 to 2009 by the US Environmental Protection Agency (EPA). I followed EPA methods to estimate reference values for specific conductivity (60 ecoregions) and each major ion (34 ecoregions) as the 25^th percentile of values in 1^st- to 4^th-order streams in Level III ecoregions with data from ≥25 sites (85 ecoregions). The 25^th percentiles of specific conductivity were <200 µS/cm for most eastern and western montane ecoregions, except those dominated by limestone or calcareous till. Arid western ecoregions had higher specific conductivities. Ca² was generally the most abundant cation followed by Mg² , Na , and K . HCO₃^- was generally the most abundant anion followed by SO₄^2- and Cl^-. Ecoregions where SO₄^2- or Cl^- concentrations were greater than HCO₃^- concentration have been affected by acidic precipitation or are influenced by marine air masses, respectively, and have very low specific conductivities. Patterns of variation appear to be associated with 3 processes controlling total and relative concentrations of major ions in freshwaters. In many ecoregions, relative ionic concentrations reflect underlying geology, but in arid ecoregions, relative ionic concentrations show concentration by evaporation. Relative ionic concentrations in coastal ecoregions and those affected by acidic precipitation reflect the ionic content of precipitation. Verification of these factors awaits better quantification of the geological and climatic characteristics of each ecoregion.