Above forest canopies, eddy covariance (EC) measurements of mass (CO2, H2O vapor) and energy exchange, assumed to represent ecosystem fluxes, are commonly made at one point in the roughness sublayer (RSL). A spatial variability experiment, in which EC measurements were made from six towers within the RSL in a uniform pine plantation, quantified large and dynamic spatial variation in fluxes. The spatial coefficient of variation (CV) of the scalar fluxes decreased with increasing integration time, stabilizing at a minimum that was independent of further lengthening the averaging period (hereafter a ‘stable minimum’). For all three fluxes, the stable minimum (CV=9–11%) was reached at averaging times (τp) of 6–7 h during daytime, but higher stable minima (CV=46–158%) were reached at longer τp (>12 h) during nighttime. To the extent that decreasing CV of EC fluxes reflects reduction in micrometeorological sampling errors, half of the observed variability at τp=30 min is attributed to sampling errors. The remaining half (indicated by the stable minimum CV) is attributed to underlying variability in ecosystem structural properties, as determined by leaf area index, and perhaps associated ecosystem activity attributes. We further assessed the spatial variability estimates in the context of uncertainty in annual net ecosystem exchange (NEE). First, we adjusted annual NEE values obtained at our long-term observation tower to account for the difference between this tower and the mean of all towers from this experiment; this increased NEE by up to 55 g C m−2 yr−1. Second, we combined uncertainty from gap filling and instrument error with uncertainty because of spatial variability, producing an estimate of variability in annual NEE ranging from 79 to 127 g C m−2 yr−1. This analysis demonstrated that even in such a uniform pine plantation, in some years spatial variability can contribute ∼50% of the uncertainty in annual NEE estimates.