Chloromethane (CH3Cl) with a global atmospheric burden of 5.3 million t is the most abundant halocarbon in the atmosphere. However, the origin of ca. 50% of the estimated annual global input of 4 million t of the gas to the atmosphere has yet to be determined. As the oceanic contribution to the global CH3Cl flux is now tightly constrained, an important terrestrial source is either underestimated or unrecognized. It has recently been proposed that higher plants may represent a CH3Cl source of sufficient magnitude to resolve the global budget imbalance. A potentially useful tool in validating CH3Cl emission flux estimates is comparison of the carbon isotope ratio of atmospheric CH3Cl with those of CH3Cl originating from various sources. Here we report the first measurements of delta13C for CH3Cl produced biologically. The CH3Cl released by the higher plant species Batis maritima and Solanum tuberosum was dramatically depleted in 13C with respect to plant tissue (delta13C = -36.8/1000 and -34.5/1000, respectively); CH3Cl released by the fungus Phellinus pomaceus also showed significant 13C depletion with respect to the wood growth substrate (delta13C = -17.9/1000). When reliable delta13C values for the other major sources of atmospheric CH3Cl become available, the distinctive isotopic signature of plant-derived CH3Cl should help constrain the contribution to the atmospheric burden from this source.