Mammalian sex hormones are spread in the environment from natural and anthropogenic sources. In the present study, the effect of estradiol on Arabidopsis thaliana growth primary metabolism, phenylpropanoid and flavonoid pathways and pathogen resistance were investigated. Treatments of Arabidopsis plants with 10 and 100 nM 17β-estradiol resulted in enhanced root growth and shoot biomass. In addition, treated plants had an increased rate of photosynthesis with a concomitant increase in carbohydrate and protein accumulation. Plants exposed to higher concentrations of 17β-estradiol (10 μM) had significantly lower root growth, biomass, photosynthesis rate, primary metabolite and phenylpropanoid and flavonoid contents indicating a toxic effect of estradiol. Treatments with increasing estradiol concentrations (10 nM, 100 nM and 10 μM) resulted in the downregulation of phenylpropanoid-flavonoid pathway genes (PAL1, PAL4, CHI and ANS) and subsequent decreased accumulation of phenolics, flavonoids and anthocyanins. Estradiol-treated plants were inoculated with Pseudomonas syringae pv. Tomato DC3000 and basal resistance was determined. Estradiol treatments rendered plants susceptible to the pathogen, thus compromising the plant defense mechanisms. These results indicate that at low concentrations, estradiol functions as a biostimulant of growth, yield and primary metabolism of Arabidopsis. However, estradiol functions as a potential transcriptional regulator of the phenylpropanoid pathway genes in Arabidopsis, having a negative effect on the phenylpropanoid and flavonoid biosynthetic pathways.