## 3 Results and Discussion  
### Antifungal In Vitro Test-Assay on Mycelium  
First, the ethanolic extract of the roots from _Magonia pubescens_ was evaluated against _Botrytis cinerea_, _Fusarium oxysporum_, and _Alternaria alternata_ (Figures 1-4 and Table S1 in Supplementary Materials), phytopathogenic fungi that affect crops worldwide, in particular, banana and tangerine crops, which are of economic relevance in Brazil. The evaluation of this ethanolic extract (EE) showed a percentage inhibition (% GI) of 30.90, 33.63, and 31.88 against _A. alternata_, _B. cinerea_, and _F. oxysporum_, respectively, at a concentration of 1 mg/mL. These results suggest some degree of antifungal activity against the tested fungi. Subsequently, in order to enrich this extract in bioactive components, the ethanolic crude extract was subjected to a liquid-liquid partition into five fractions, hexanes (F-H), dichloromethane (F-D), ethyl acetate (F-E), _n_-butanol (F-B), and water (F-W) fractions. A preliminary Thin Layer Chromatography (TLC) analysis of these fractions revealed similar phytochemical profiles of hexanes and dichloromethane fractions, and these were combined (F-H/D). These fractions were further evaluated against the three phytopathogenic fungi. The extract and fractions were first tested at a concentration of 1 mg/mL, and those exhibiting growth inhibition higher than 20% were tested at lower concentrations, namely, 0.5 and 0.1 mg/mL. Fosbel Plus was used as a reference fungicide [39], whereas ethanol was used as a negative control, using one dish per pathogen and eight disks for each control. Results obtained on _A. alternata_ (Figure 1) revealed that fractions F-H/D and F-B (% GI 27.10 and 29.35, respectively) showed similar potency to the crude extract (% GI 30.90), whereas the F-E and F-W fractions were inactive (GI < 10% at 1 mg/mL concentration). Thus, this procedure resulted in maintaining the activity of the two organic fractions, hexanes/dichloromethane and _n_-butanol fractions, in comparison with the ethanolic extract at all the assayed concentrations but did not result in an enrichment in bioactive components. Figure 1: Antifungal effects (% growth inhibition, % GI) of the ethanolic extract (EE) and fractions (F-H/D: hexane/dichloromethane and F-B: _n_-butanol) from _M. pubescens’_ roots against _A. alternata_. Fosbel Plus was used as a positive control. Results are expressed as a percentage relative to the negative control. Data are presented as means \(\pm\) SD (standard deviation, \(n\) = 8); \(p\) < 0.05 (*). F-E (ethyl acetate) and F-W (water) fractions were inactive (GI < 10%). Extract/fractions with a GI > than 20% at 1 mg/mL were assayed at lower concentrations (0.5 and 0.1 mg/mL). Likewise, antifungal activity of the ethanolic extract and fractions on B. cinerea (Figure 2) led to a similar effect of the F-H/D fraction (% GI 31.74 at 1 mg/mL) and a slight increase in activity of the F-B fraction (% GI 39.18 at 1 mg/mL) compared to the ethanolic extract (% GI 33.63 at 1 mg/mL). However, the F-E and F-W showed low potency to inactivity with respect to the EE. _F. oxysporum_ (Figure 3) showed a similar behavior to the extract and fractions as the other two assayed fungi. Thus, the F-H/D (% GI 28.34) and F-B (% GI 32.15) fractions exhibited similar effects as the ethanolic extract (% GI 31.88), whereas the F-E and F-W fractions were almost inactive (GI around or lower than 10% at 1 mg/mL). Figure 3: Antifungal effects (% growth inhibition) of the ethanolic extract (EE) and fractions (F-H/D-hexane/dichloromethane, F-E: ethyl acetate fraction, and F-B: _n_-butanol) from _M. pubescens_’ roots against _F. oxysporum_. Results are expressed as a percentage relative to the negative control. Fosbel Plus was used as a positive control. Data are presented as means \(\pm\) SD (standard deviation, \(n\) = 8); \(p\) < 0.05 (*). F-W (water) fraction was inactive (GI < 10%). Extract/fractions with a GI > than 20% at 1 mg/mL were assayed at lower concentrations (0.5 and 0.1 mg/mL).