Rapid changes in climate may impose strong selective pressures on organisms. Evolutionary responses to climate change have been observed in natural populations, yet no example has been documented for a metabolic enzyme locus. Furthermore, few studies have linked physiological responses to stress with allozyme genotypic variation. We quantified changes in allele frequency between 1988 and 1996 at three allozyme loci (isocitrate dehydrogenase, Idh; phosphoglucose isomerase, Pgi; and phosphoglucomutase, Pgm) for the leaf beetle Chrysomela aeneicollis in the Bishop Creek region of the Sierra Nevada of California (2900-3300 m). Beetles often experience high daytime (> 32 degrees C) and extremely low nighttime (< -5 degrees C) temperatures during summer. Bishop Creek weather station data indicated that conditions were unusually dry before 1988, and that conditions were cool and wet during the years preceding the 1996 collection. We found directional changes in allele frequency at Pgi (11% increase in the Pgi-1 allele), but not at Idh or Pgm. We also found that physiological response to thermal extremes depended on Pgi genotype. Pgi 1-1 individuals induced expression of a 70-kD heat shock protein (HSP) at lower temperatures than 1-4 or 4-4 individuals, and 1-1 individuals expressed higher levels of HSP70 after laboratory exposure to temperatures routinely experienced in nature. Survival after nighttime laboratory exposure to subzero temperatures depended on gender, previous exposure to cold, and Pgi genotype. Females expressed higher levels of HSP70 than males after exposure to heat, and recovery by female Pgi 1-1 homozygotes after exposure to cold (-5 degrees C) was significantly better than 1-4 or 4-4 genotypes. These data suggest that the cooler climate of the mid-1990s may have caused an increase in frequency of the Pgi-1 allele, due to a more robust physiological response to cold by Pgi 1-1 and 1-4 genotypes.