Background and Aims: The largest diversity of the genus Vanilla shows a preference for humid and flooded areas such as palm-swamp peat forests, which limits the collection and quality of specimens. This study focuses on the vegetative propagation of Vanilla pompona subsp. grandiflora in flood-prone areas of the Alto Mayo Valley, Peru. The objective is to determine the optimal number of nodes for the healthy growth of cuttings, using low-cost substrates. Methods: The research was conducted in the nursery of Finca Don Pepito in Tingana, department of San Martín, Peru. A fully randomized experimental design was employed, considering four types of cuttings and four substrates (treatments). Observations were made throughout the experiment, assessing bud activation, the health of the cuttings, and their relationship between the treatments. Key results: The results indicate that bud activation is significantly influenced by the number of nodes in the cuttings, showing greater efficiency as the number of nodes increases. Conclusions: Propagation through two-node cuttings is considered the best alternative due to the limited availability of genetic material. Furthermore, the health of the cuttings is more closely related to the type of cutting rather than to the substrate treatments. © 2024 Elsevier B.V., All rights reserved.