Ethylene seems to play a secondary role in non-climacteric strawberry ripening compared to abscisic acid. However, this does not exclude that ethylene can regulate some specific events related to the ripening process. Preliminary experiments of applications of ethylene or its inhibitor 1-MCP to strawberry fruits have reinforced this hypothesis. Here, we reveal some previously non-covered physiological effects of ethylene using an in vitro strawberry ripening system. Fruits of Fragaria chiloensis treated with ethephon at the large green developmental stage showed inhibition of anthocyanin biosynthesis and downregulation of essential anthocyanin biosynthesis genes during the ripening. At the same time, ethylene stimulated lignin biosynthesis and remarkably upregulated the expression of FcPOD27. Since contrasting results have been reported when ethylene was applied at late ripening developmental stages, our findings support the hypothesis of a temporal-specific ethylene role in the ripening of strawberry fruits.