Dark sericitic material at and near the top of the 2.765 ± 0.01 Ga Mount Roe #2 paleosol in Western Australia contains 0.05–0.10 wt% organic carbon with δ13C values between −33‰ and −51‰ PDB (Peedee belemnite). Such negative isotopic values strongly indicate that methanotrophs once inhabited this material. The textures and the chemical composition of the dark sericitic material indicate that the methanotrophs lived in or at the edges of ephemeral ponds, that these ponds became desiccated, and that heavy rains transported the material to its present sites. The discovery of methanotrophs associated with the Mount Roe #2 paleosol may extend their geologic record on land by at least 1.5 b.y. Methanotrophy in this setting is consistent with the notion that atmospheric methane levels were ≥ 20 µatm during the Late Archean. The radiative forcing due to such high atmospheric methane levels could have compensated for the faint younger sun and helped to prevent massive glaciation during the Late Archean.