The positive effect of silicon on plants is thought to be mediated by a modification of phenolic metabolism. The purpose of the study was to evaluate the effect of a silicon-based mechanocomposite (MC) on alterations of the phenolic profile of strawberry plants in the course of development under in vitro, ex vitro, and in vivo conditions. Aqueous ethanol extracts of aboveground parts of in vitro–derived plants (Fragaria × ananassa cv. ‘Solnechnaya polyanka’) were subjected to HPLC. Nineteen individual phenolic compounds (hydroxybenzoic and hydroxycinnamic acids, catechins, ellagic acid derivatives, and flavonol glycosides) were quantified. The results revealed phenolic profiles specific to each studied stage and significant transformations of the profiles by the MC. It induced strong upregulation of hydroxycinnamic acid during in vitro rooting and of catechins and hydroxybenzoic acids during ex vitro acclimation. At ex vitro and in vivo stages, the emergence of quercetin glycosides and ellagitannins was registered, and the MC elevated their levels during ex vitro acclimation and field growth. Principal component analysis confirmed the significant effect of the MC on the phenolic profile at all stages, and this effect was the strongest during ex vitro acclimation. The results are consistent with previous reports on the modification of phenolic profiles of plants by silicon-derived biostimulants.