## 5 Conclusions  
This research is the first carried out on Italian _E. orgzicola_ focusing on the miRNA-mediated epigenetic regulation of genes involved in profoxydim resistance.  
Our findings highlight that herbicide treatment can trigger miRNA transcription, causing a down-regulation in the expression of target genes (i.e., _ata-miR166c-5p_ vs. _CYP72A122_, _ath-miR847_ vs. _CYP71 AK2_, _osa-miR85538_ vs. _EcGST_, and _gra-miR7487c_ vs. _CYP72A254_). These results are a basis for further investigations of epigenetic regulation mediated by miRNAs as part of studies aimed at obtaining more information on the mechanisms of herbicide resistance.  
At present, the entire miRNAome of _E. orgzicola_ is not available. The availability of a large set of miRNAs able to affect herbicide detoxification would be useful to provide deeper insights into the complex mechanisms underlying the resistance phenomenon.  
The methodological analysis of our present study represents an innovative approach to assessing epigenetic mechanisms affecting herbicide resistance and could be applied to other crops, such as in vineyards, where, in recent years, the evolution of weeds resistant to the most commonly used herbicides (i.e., glyphosate) has been observed.  
A better understanding of the epigenetic regulation of herbicide resistance will be useful for planning more targeted and sustainable methods for controlling this phytosanitary threat, reducing inputs, and optimizing precision weed management (PWM) technologies. Moreover, it can be hypothesized that miRNAs could be applied as alternative biopesticides in modern agriculture due to their ability to migrate among organisms and regulate specific genetic processes.  
Our results highlighted the miRNA-mediated post-transcriptional regulation of the expression of genes responsible for resistance. Since miRNAs are known to be influenced by ecological factors, conducting tests under controlled conditions to evaluate which biotic and abiotic factors (i.e., climate change, edaphic properties, soil microbial communities) may play a role in influencing miRNA-mediated resistance would be a favorable strategy to obtain new information that is useful for predicting the occurrence of resistance, favoring a proactive approach to handle this phytosanitary threat.  
Conceptualization, C.M.C., E.C., A.M.P. and M.B.; methodology, C.M.C., E.C., A.M.P., M.G., E.G., P.Z. and M.B.; validation, E.C. and M.B.; fieldwork, C.M.C.; formal analysis, C.M.C., E.C. and M.B.; investigations, C.M.C., E.C. and M.B.; resources, C.M.C., E.C., A.M.P. and M.B.; data curation, C.M.C., E.C. and M.B.; writing--original draft preparation, C.M.C., E.C. and M.B.; writing--review and editing, E.C., A.M.P. and M.B.; visualization, C.M.C., E.C. and M.B.; supervision, E.C. and M.B.; project administration, M.B.; funding acquisition, M.B. All authors have read and agreed to the published version of the manuscript.  
This publication is part of the project NODES, funded by the European Union--NextGenerationEU, Mission 4 Component 1.5--ECS0000036--CUP F17G22000190007.  
The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.  
The authors wish to thank [PERSON], [PERSON], and the team of the Lombardy Region Plant Protection Service Laboratory; [PERSON] and [PERSON] of the Department of Earth and Environmental Sciences of the University of Pavia; [PERSON], [PERSON], [PERSON], and [PERSON] of Innova-Tech S.r.l.; [PERSON] with the team of Agri.Bio; [PERSON] for their valuable help.  
Authors [PERSON], [PERSON], [PERSON] were employed by the company Agricola 2000 S.c.p.A. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.