We determined rates of acetylene reduction and estimated total nitrogen fixation associated with bryophytes, lichens, and decaying wood in Hawaiian montane rain forest sites with underlying substrate ranging in age from 300 to 4.1 million years. Potential N fixation ranged from ca 0.2 kg/ha annually in the 300-year-old site to ca 1 kg/ha annually in the 150,000-year-old site. Rates of acetylene reduction were surprisingly uniform along the soil-age gradient, except for high rates in symbiotic/associative fixers at the 150,000-year-old site and in heterotrophic fixers at the 2100-year-old site. Low fixation at the youngest site, where plant production is known to be N-limited, suggests that demand for N alone does not govern N fixation. Total N fixation was highest in sites with low N:P ratios in leaves and stem wood, perhaps because epiphytic bryophytes and lichens depend on canopy leachate for mineral nutrients and because heterotrophic fixation is partly controlled by nutrient supply in the decomposing substrate; however, differences in substrate cover, rather than in fixation rates, had the largest effect on the total N input from fixation at these sites.