Yeasts form mutualistic interactions with insects. Hallmarks of this 24 interaction include provision of essential nutrients, while insects facilitate yeast 25 dispersal and growth on plant substrates. A phylogenetically ancient, chemical 26 dialogue coordinates this interaction, where the vocabulary, the volatile chemicals 27 that mediate the insect response, remains largely unknown. Here, we employed gas 28 chromatography-mass spectrometry (GC-MS), followed by hierarchical cluster 29 (HCA) and orthogonal partial least square discriminant analysis (OPLS-DA), to 30 profile the volatomes of six Metschnikowia spp., Cryptococcus nemorosus and 31 brewer's yeast Saccharomyces cerevisiae. The yeasts, which are all found in 32 association with insects feeding on foliage or fruit, emit characteristic, species-33 specific volatile blends that reflect the phylogenetic context. Species-specificity of 34 these volatome profiles aligned with differential feeding of cotton leafworm larvae 35 Spodoptera littoralis on these yeasts. Bioactivity correlates with yeast ecology; 36 phylloplane species elicited a stronger response than fruit yeasts, and larval 37 discrimination may provide a mechanism for establishment of insect-yeast 38 associations. The yeast volatomes contained a suite of insect attractants known from 39 plant and especially floral headspace, including (Z)-hexenyl acetate, ethyl (2E,4Z)-40 deca-2,4-dienoate (pear ester), (3E)-4,8-dimethylnona-1,3,7-triene (DMNT), linalool, 41 α-terpineol, β-myrcene or (E,E)-a-farnesene. A wide overlap of yeast and plant 42 volatiles, notably floral scents further emphasizes the prominent role of yeasts in 43 plant-microbe-insect relationships including pollination. The knowledge of insect-44 Ljunggren et al. -p. 3 yeast interactions can be readily brought to practical application, live yeasts or yeast 45 metabolites mediating insect attraction provide an ample toolbox for the 46 development of sustainable insect management. 47 IMPORTANCE Yeasts interface insect herbivores with their food plants. 48 Communication depends on volatile metabolites, and decoding this chemical 49 dialogue is key to understanding the ecology of insect-yeast interactions. This study 50 explores the volatomes of eight yeast species which have been isolated from foliage, 51 flowers or fruit, and from plant-feeding insects. They each release a rich bouquet of 52 volatile metabolites, including a suite of known insect attractants from plant and 53 floral scent. This overlap underlines the phylogenetic dimension of insect-yeast 54 associations, which according to the fossil record, long predate the appearance of 55 flowering plants. Volatome composition is characteristic for each species, aligns with 56 yeast taxonomy, and is further reflected by a differential behavioural response of 57 cotton leafworm larvae, which naturally feed on foliage of a wide spectrum of broad-58 leaved plants. Larval discrimination may establish and maintain associations with 59 yeasts and is also a substrate for designing sustainabl...