Background: Being a promising tropical woody oilseed crop, the evergreen and recurrent plants of Sacha Inchi (Plukenetia volubilis L.) has complex phenology and source–sink interactions. Carbon source–sink manipulations with control and two treatments (reduce source, ca. 10% mature leaf pruning; reduce sink, 10% fruitlet thinning) were conducted on 2.5-year-old field-grown P. volubilis plantation during the early-wet season in a seasonal tropical area.Results: Leaf photosynthetic rate and specific leaf area largely remained unchanged in response to defoliation or defloration. Compared with control, higher N contents on average were observed in both remaining leaves and branches of the defoliated plants, suggesting that N-mobilization was mainly due to the enhanced N uptake from soil. Carbon, but not N, is a source-driven growth process of P. volubilis plants, as defoliation reduced the contents of non-structural carbohydrates (especially sugar) in branches, although temporally, whereas defloration increased available C reserve. The seasonal dynamic pattern of fruit ripening was altered by source–sink regulations. The total seed yield throughout the growing season that depends on fruit set and retention (i.e., number of matured fruit) rather than individual fruit development (size), was slightly increased by defloration but was significantly decreased by defoliation. Compared with control, defloration did not enrich the KEGG pathway, but defoliation downregulated the TCA cycle and carbohydrate and lipid metabolisms in fruitlets after 24 days of the applications of source–sink manipulation. Conclusion: Carbohydrate reserves served to buffer sink–source imbalances that may result from temporary adjustment in demand for assimilates (e.g., defloration) or shortfalls in carbon assimilation (e.g., defoliation). Defoliation is disadvantageous for the yield and also for carbohydrate and lipid accumulation in fruits of P. volubilis plants. Although more studies are needed, these results provide new insights to the further improvement in seed yield of the strong source-limited P. volubilis plants by source/sink manipulations.