## 3 Discussion
In fact, it was reported that the overexpression of _CYP81A12_ and _CYP81A21_ confers cross-resistance to ALS (bensulfuron-methyl and penoxsulam) and ACCase (diclofop-methyl, tralkoxydim, pinoxaden) in _E.phylopogon_, while _CYP81A24_ is involved in the detoxification of cyclohexanedione (ACCase inhibitor), such as profoxydim. Furthermore, [PERSON] et al. (2020) highlighted that in _E.phylopogon_, _CYP81A12_, _CYP81A21_, and _CYP81A24's aminoacidic sequences are similar [13]. This fact would explain the synergistic action of these three proteins towards profoxydim. [PERSON] et al. (2019) also reported how _CYP81A63_ was involved in the metabolization of ACCase inhibitors in barley, cyclohexanedione included [17]. In our experimental conditions, we assessed that the transcription of some miRNAs (_osa-miRR166a-5p, osa-miRR166d-5p, osa  
Figure 3: Radar chart of the expression profiles of genes (blue line) and miRNAs (red line) in the SUS biotype (**A**), in the RESI biotype (**B**), and in the RESI biotype (**C**) after profoxydim treatment. ata—_Aegilops tauschii_ Coms. ; ath—_Arabidopsis thaliana_ (L) [PERSON]; gma–_Glycine max_ (L) [PERSON]; gra–_Gossspium raimondii_ Ulbr. ; osa—_Oryza sativa_ L.  
miR2099-5p_, and _osa-miR396a-5p_), selected for their ability to target the abovementioned genes, was not triggered by profoxydim, allowing the overexpression of cytochromes P450. [PERSON] et al. (2014) observed the overexpression (i.e., higher metabolizing activity) of _CYP7 LAK2_, _CYP72A122_, and _CYP7A254_ in _E.phylopogon_ after treatment with bispyibracNa (ALS inhibitor) [21]. In our present study, we found that _gma-miR396f_, _ata-miR166c-5p_, and _gra-miR7486c_ targeting these cytochromes P450 were triggered following profoxydim application, causing a down-regulation of gene expression. Overexpression of _GSTF1_ and _elF4B_ and reduced transcription of _gra-miR8759_ and _osa-miR5538_ were observed after herbicide treatment. Our findings are in agreement with those of [PERSON] et al. (2018) [23] and [PERSON] et al. (2013) [20], who reported an increased expression of the same genes in _E. crus-galli_ resistant to imazetheapyr (ALS inhibitor), as well as those of [PERSON] et al. (1999, 2003, 2013) [47; 48; 49], who observed an increased expression of GST genes in _Alopecurus myosuroides_ Huds. and _Lolium rigidum_ Gaudin resistant to chlorotoluron (inhibitor of photosynthesis at photosystem II) and fenoxaprop-P-ethyl (ACCase inhibitor). After treatment with profoxydim, we observed a reduced expression of _EcGST_, with an increased expression of _osa-miR395f_ in the RES biotypes, suggesting the ability of this miRNA to down-regulate glutathione S-transferase. In our study, we considered target genes that are known to be involved in resistance in _Echinochloa_ and are widely studied. Next, we searched in silico for miRNA sequences with a high matching rate with the target genes. The predicted miRNAs were finally validated in _E. orgzicola_ by means of RT-qPCR. Moreover, we highlighted their role in the down-regulation of some genes involved in profoxydim resistance. All of the miRNAs tested in this research and previously reported in other plant species [50; 51; 52; 53; 54; 55; 56; 57] were also expressed in _E. orgzicola_. Moreover, the increased expression of _ata-miR166c-5p_, _ath-miR8847_, _osa-miR85538_, and _gra-miR7487c_, along with the reduced expression of _CYP7A2122_, _CYP7 LAK2_, _EcGST_, and _CYP7A254_, suggests the occurrence of a miRNA-mediated down-regulation of these detoxifying enzymes triggered by profoxydim. Since it is known that the transcription of sncRNAs is influenced by ecological factors [27; 28; 29; 30], our present findings provide the basis for future insights into the relationship between ecological factors and resistance occurrence. Planning new and sustainable control strategies aimed at reducing pesticide inputs also requires the assessment of the entire miRNAome of _E. orgzicola_ in order to better understand the epigenetic regulation of proteins involved in herbicide resistance.