## 3 Results and Discussion
### Insect Antifeordant Activity against Chrysodeixis chalcites  
The ethanolic extract and fractions from _M. pubescens_' roots were evaluated on _Chrysodeixis chalcites_, an insect that causes severe damage to crops worldwide. In this assay, samples were dissolved in ethanol, and 5 \(\mu\)L from a stock solution of 40 mg/mL per disk 0.2 mg/cm\({}^{2}\) were used. Indoxacarb 30% (Steward 30 WG), a frequently used synthetic insecticide, was used as a positive control, whereas a solvent was used as a negative control. Under the experimental conditions described in the previous section, in the choice assay and the non-choice assays, three five-instar larvae of _C. chalcites_ were placed in each petri dish (six replicates for each experiment), and after a 20 h period, the ingested leaf area was measured, and the refusal rate (FR) was calculated. A value of FR > 50% in the choice or non-choice assays indicates a significant feeding inhibition [44]. The combination of FR values > 50% at a dose of 0.2 mg/cm\({}^{2}\) in both assays indicates a strong antifeedant effect. Samples tested with a combination of FR < 50% were considered inactive (Table 1, and Figure 5). \begin{table}
\begin{tabular}{c c c c} \hline \hline
**Sample 2** & \multicolumn{2}{c}{**Choice Feeding Assay**} & **Non-Choice Feeding Assay** & **Antifeordant Category** \\ \hline EE 3 & 92.14 & 95.16 & strong insect antifeedant \\ F-H/D 3 & 60.52 & \(-\)27.67 & weak insect antifeedant \\ F-E 3 & \(-\)108.09 & 61.10 & appetite suppressor \\ F-B & \(-\)6.69 & 60.71 & appetite suppressor \\ F-W 3 & \(-\)74.57 & 44.73 & inactive \\ C 4 & 97.95 & 99.04 & strong insect antifeedant \\ \hline \hline \end{tabular}  
* (% Treated disks consumed/% Control disks consumed)] \(\times\) 100.
* Samples were dissolved in ethanol: 5 \(\mu\)L per disk-0.2 mg/cm\({}^{2}\). 3 EE: ethanolic extract, F-H/D hexane-dichloromethane fraction, F-E: ethyl acetate fraction, FB: \(n\)-butanol fraction, F-W: water fraction. 4 C: indoxacarb 30% (STEWARD® 30 WG) was used as a positive control.  
\end{table} 
Table 1: Insect antifeeding activity (% FR) 1 of the ethanolic extract and fractions from _Magonia_ _pubescens_’ roots against _Chrysodeixis chalcites_. Figure 4: Petri dishes of the in vitro test-assay on _A. alternata, B. cinerea_, and _F. oxysporum_. The results indicate that the crude extract (EE) had a FR > 90% in the choice and non-choice assays, meaning that the extract acts very effectively as an antifeedant for _C. chalcites_ larvae. Subsequently, hexane/dichloromethane (F-H/D), ethyl acetate (F-E), _n_-butanol (F-B), and water (F-W) fractions obtained by liquid-liquid partition of the ethanolic crude extract were further evaluated against _C. chalcites_. The results revealed that the hexane/dichloromethane fraction (F-H/D) acts as a weak insect antifeedant, since in the choice assay, larvae preferred to feed on untreated disks (FR = 60.52%), indicating good activity, but in the non-choice assay, the larvae fed from the treated disks (FR = \(-27.67\%\)), so it does not have a complete antifeedant effect. Moreover, the ethyl acetate (F-E) and _n_-butanol (F-B) fractions showed positive FR values in the non-choice assay (FR 61.10% and 60.71%, respectively), though activity was not confirmed by the choice assay (low FR values). Thus, the F-E and F-B fractions are appetite suppressants since larvae do not feed off treated disks in the non-choice assay. On the other hand, the water (F-W) fraction was inactive, showing FR values of \(-74.57\) and \(44.73\%\), respectively, in the assays. Notably, the crude ethanolic extract showed similar effects as Steward 30 WG [42], the commercial insecticide used as a positive control, exhibiting an FR of 92.14% and 95.16% in the choice and non-choice feeding assays, respectively. This fact suggests that the plant could be used, after a simple and accessible procedure, as a biopesticide against this phytopathogenic insect. The only study regarding the potential insecticide activity of _M. pubescens_ is by [PERSON] et al.