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Abstract. Decaying vegetation was determined to be a potentially important source ofatmospheric ice nucleation particles (INPs) in the early 1970s. The bacteriumPseudomonas syringae was the first microorganism with ice nucleationactivity (INA) isolated from decaying leaf litter in 1974. However, the icenucleation characteristics of P. syringae are not compatible withthe characteristics of leaf litter-derived INPs since the latter were foundto be sub-micron in size, while INA of P. syringae depends on muchlarger intact bacterial cells. Here we determined the cumulative icenucleation spectrum and microbial community composition of the historic leaflitter sample 70-S-14 collected in 1970 that conserved INA for 48 years. Themajority of the leaf litter-derived INPs were confirmed to be sub-micron insize and to be sensitive to boiling. Culture-independent microbial communityanalysis only identified Pseudomonas as potential INA.Culture-dependent analysis identified one P. syringae isolate, twoisolates of the bacterial species Pantoea ananatis, and one fungalisolate of Mortierella alpina as having INA among 1170 bacterialcolonies and 277 fungal isolates, respectively. Both Pa. ananatisand M. alpina are organisms that produce heat-sensitive sub-micronINPs. They are thus both likely sources of the INPs present in sample 70-S-14and may represent important terrestrial sources of atmospheric INPs, aconclusion that is in line with other recent results obtained in regard toINPs from soil, precipitation, and the atmosphere.