1–Methylcyclopropene (1–MCP) is used for extending the postharvest life of the avocado during storage. Evaluated the effect of 1–MCP application at different times after harvest, i.e., 0, 7, 14, and 21 d at 5◦ C, to identify the threshold of the ethylene inhibition response in “Hass” avocado. Our results showed that fruits from two maturity stages at harvest: low dry matter (20–23%) and high dry matter (27%). Changes in ethylene production rates and transcript accumulation of genes involved in ethylene metabolism were measured at harvest and during storage. 1–MCP treated fruit up to 14 d of storage showed similar values of firmness and skin color as fruit treated at harvest time. In contrast, when the application was performed after 21 d, the fruit showed ripening attributes similar to those of the untreated ones. To further understand the molecular mechanisms responsible for the lack of response to 1–MCP at 21 d of storage, transcriptomic analysis was performed. Gene ontology analyses based on the DEG analysis showed enrichment of transcripts involved in the ‘response to ethylene’ for both maturity stages. All genes evaluated showed similar expression profiles induced by cold storage time, with a peak at 21 d of storage and an increased softening of the fruit and peel color. This was a two-year field study, and results were consistent across the two experimental years. Our results should help growers and markets in selecting the optimal timing of 1–MCP application in “Hass” avocados and should contribute to a deeper understanding of the molecular mechanisms of the avocado ripening process.