The objective of this study was to evaluate the antioxidant capacity by spectrophotometric methods, the in vitro and in vivo antifungal effect against Lasiodiplodia theobromae and the constitution of the essential oils (EO) of oregano and thyme in comparison with their commercial counterparts. The results showed by the EOs of extracted thyme (T-EO), commercial thyme (CT-EO), extracted oregano (O-EO) and commercial oregano (CO-EO), demonstrated antioxidant profiles with a radical neutralizing potential (DPPH•) of IC50: 1.11 ± 0.019; 1.08 ± 0.05; 40.56 ± 0.227 and 0.69 ± 0.004 mg/mL, respectively. They also revealed a ferric ion reducing capacity (FRAP) of 93.05 ± 0.52; 97.72 ± 0.42; 21.85 ± 0.57 and 117.24 ± 0.64 mg Eq Trolox/g. A reduction in β-carotene degradation of 65.71 ± 0.04; 51.97 ± 0.66; 43.58 ± 1.56 and 57.46 ± 1.56 %. A total phenol content (Folin-Ciocalteu) of 132.97 ± 0.77; 141.89 ± 2.56; 152.04 ± 0.10 and 25.66 ± 0.40 mg EGA/g. Chemical characterization performed by gas chromatography mass spectrometry (GC-MS) showed that the respective major components of the samples were thymol (T-EO: 45.78 %), thymol (CT-EO: 43.57 %), alloaromadendrene (O-EO: 25.17 %) and carvacrol (CO-EO: 62.06 %). Regarding antifungal activity, it was evident that at the in vitro level, both commercial EOs had a MIC of 250 ppm while the extracted thyme EO had a MIC of 500 ppm; In vivo studies demonstrated that the application of thyme EO had a behavior similar to the synthetic fungicide, slowing down rot in bananas under storage conditions. Finally, partial least squares discriminant analysis (PLS-DA) and heat maps suggest p-cymene, carvacrol, linalool, eucalyptol, 4-terpineol, (z)-β-terpineol, alkanhol, caryophyllene, β-myrcene, d-limonene, α-terpinene, α-terpineol, d-α-pinene, camphene, caryophyllene oxide, δ-cadinene, terpinolene and thymol as relevant biomarkers associated with the assessed bioactive properties demonstrating the potential of extracted essential oils for the development of a botanical biofungicide.