Novel plant assemblages are a long-recognized feature of late-glacial North America, but identifying their causes has been hampered by inaccurate radiocarbon chronologies and the multiplicity of ecological and climatic events during the late Pleistocene. Recently we reported that the formation of no-analog vegetation may have been linked to declines in Pleistocene megafaunal communities, based on pollen and spores from the coprophilous fungus Sporormiella at sites in Indiana and New York. We present a new, multi-proxy analysis from Silver Lake, OH, which 1) updates the radiocarbon chronology of a classic pollen record with a well-established zone of no-analog vegetation, 2) combines a new sub-centennial pollen record with charcoal, Sporormiella, and x-ray fluorescence (XRF) spectroscopy analyses for an integrated record of landscape change before, during, and after the period of no-analog vegetation, and 3) replicates both the absolute and relative temporal patterns of landscape change at Appleman Lake, IN. At Silver Lake, the decline in Sporormiella at 13.9 ka BP was immediately followed by the formation of novel plant assemblages, as well as the highest-magnitude charcoal peak in the record. Increased Ca and Sr concentrations during the no-analog interval indicate either increased moisture, increased input of nutrients from deciduous litter, or both. The duration of the no-analog assemblages (13.9–11.8 ka BP) roughly corresponds to the period of peak insolation dissimilarity, but is more temporally constrained than previously reported in subcontinental-scale syntheses (17–11 ka BP). We propose a hierarchy of controls on late-glacial plant communities, where biotic interactions such as megaherbivory mediate climate-driven vegetation change.