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Colleters are secretory structures involved in the protection of young and developing plant organs. Although the presence of colleters in Gentianales is described as a synapomorphy, studies on the morphofunctionality of colleters and the mechanisms underlying the synthesis and release of colleter secretion in Gentianaceae are scarce. Here, we described the ontogeny and the morphological and functional aspects of colleters of Prepusa montana, revealed the nature of the key compounds present in the secretion, and explored the cellular aspects of the synthesis and release of secretion and senescence of colleters. Samples of the stem apical meristem with leaf primordium and young leaves; adult and senescent leaves were observed using light and electron microscopy. The colleters, located in the axil of the leaf, have a protodermal origin and develop asynchronously. They are digitiform, possessing a short peduncle and a secretory head containing homogeneous cells with dense cytoplasm and abundant endoplasmic reticulum and Golgi bodies. The secretion, composed of polysaccharides and proteins, is accumulated in schizogenous spaces and released through the separation of peripheral secretory cells and loosening of the external periclinal wall. Presumably, senescence is caused by programmed cell death. The morphoanatomical characterization of P. montana leaf colleters described here is the first record for the genus and the peculiar accumulation of colleter secretion in schizogenous spaces expanding our knowledge on the diversity of these secretory structures. Our results also provide insights into programmed cell death as an eminent topic related to secretory structures., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)