<p>Fusarium wilt of strawberry (FWS), caused by <i>Fusarium oxysporum</i> f. sp.<i> fragariae</i> (Fof) can greatly reduce yield in strawberries, and alternatives for its management are needed. The hypothesis that strawberry plants treated with native antagonistic strains and biochar nutrition (BOC) could increase their resistance against FWS was investigated. The study was conducted under greenhouse conditions; Fof was inoculated in the soil, and bentonite–vermiculite bioformulation containing a mixture of <i>Trichoderma</i> strains (TASMix) was applied as a soil treatment. BOC was applied as a soil amendment at a rate of 3% (w/w) one day before transplanting. The results showed that incorporating TASMix in soil substrate containing BOC showed excellent control efficacy compared to separate treatments. This treatment reduced wilt incidence and wilting/stunting and vascular discoloration/rotting severities by 49.4%, 55.9%, and 57.2%, respectively, compared with the non-treated control ones. Also, the decrease in strawberry plants’ growth due to the pathogen-induced stress was synergistically reduced. Quantitative PCR revealed that applying TASMix or BOC alone significantly reduced the pathogen abundance in rhizosphere soil. Nonetheless, the integrated treatment (TASMix + BOC) resulted in the lowest Fof abundance in strawberry rhizosphere soil, measuring − 4.82 log₁₀ ng DNA g^− 1. The results showed that the BOC also positively affected <i>Trichoderma</i> rhizospheric colonization. Importantly, TASMix + BOC treated plants exhibited faster and stronger systemic responses of the defense genes <i>FaBG2-2</i> and <i>FaCHI3-1</i>. These results suggested a strategy based on using <i>Trichoderma</i> inoculums in combination with biochar can be proposed for the integrated management of FWS on organic strawberry cultivations.</p>