Resistance to acetolactate synthase (ALS) inhibiting herbicides has recently been reported in Glebionis coronaria from wheat fields in northern Tunisia, where the weed is widespread. However, potential resistance mechanisms conferring resistance in these populations are unknown. The aim of this research was to study target-site resistance (TSR) and non-target-site resistance (NTSR) mechanisms present in two putative resistant (R) populations. Dose–response experiments, ALS enzyme activity assays, ALS gene sequencing, absorption and translocation experiments with radiolabeled herbicides, and metabolism experiments were carried out for this purpose. Whole plant trials confirmed high resistance levels to tribenuron and cross-resistance to florasulam and imazamox. ALS enzyme activity further confirmed cross-resistance to these three herbicides and also to bispyribac, but not to flucarbazone. Sequence analysis revealed the presence of amino acid substitutions in positions 197, 376, and 574 of the target enzyme. Among the NTSR mechanisms investigated, absorption or translocation did not contribute to resistance, while evidences of the presence of enhanced metabolism were provided. A pretreatment with the cytochrome P450 monooxygenase (P450) inhibitor malathion partially synergized with imazamox in post-emergence but not with tribenuron in dose–response experiments. Additionally, an imazamox hydroxyl metabolite was detected in both R populations in metabolism experiments, which disappeared with the pretreatment with malathion. This study confirms the evolution of cross-resistance to ALS inhibiting herbicides in G. coronaria from Tunisia through TSR and NTSR mechanisms. The presence of enhanced metabolism involving P450 is threatening the chemical management of this weed in Tunisian wheat fields, since it might confer cross-resistance to other sites of action.

Glebionis coronaria (L.) Cass. ex Spach is a troublesome weed in cereal cropping systems in northern Tunisia. Recently, failures in controlling this weed have been reported by farmers. Field surveys and farmers interviews were conducted to highlight the potential causes of G. coronaria occurrence and the associated yield losses in wheat. Survey results revealed a significant correlation between farmers’ awareness of resistance occurrence and cultural practices, mainly sowing date and tillage, while G. coronaria abundance was related to the lack of herbicide rotation and the frequency of ALS-inhibiting herbicide use. High G. coronaria infestations (more than 20 plants/m2) caused a significant decrease in wheat grain yield, reaching almost 75% at a density of 100 plants/m2. Field and pot experiments showed low efficacies of ALS-inhibiting herbicides to control G. coronaria populations. The application of field rates of tribenuron-methyl and mesosulfuron + iodosulfuron failed to control the tested populations, and generally, G. coronaria dry weight increased compared to nontreated ones (potential hormetic effect). These findings were further investigated in two selected resistant populations through tribenuron dose–response experiments, plants from both populations exhibited high resistance factors (greater than 300), surviving up to 16-fold the recommended field dose. This is the first report for G. coronaria resistance occurrence to ALS-inhibiting herbicides in Tunisia and the second case worldwide.

Glebionis coronaria (L.) Cass. ex Spach is one of the most serious weeds in cereal crops in Northern Tunisia. Our previous studies have confirmed the presence of resistance to ALS-inhibiting herbicides in G. coronaria, showing the evolution of cross-resistance through TS and NTS mechanisms. The purpose of this study was to evaluate the effects of (i) two tillage practices (conventional tillage and reduced tillage), (ii) three known allelopathic crops: Hordeum vulgare L. (Barley), Brassica napus L. (Rapeseed) and Triticum durum (Wheat) and (iii) herbicides belonging to different modes of action on the density, the cohort numbers and the growth of an ALS-inhibiting cross-resistant population of G. coronaria. Field experiments were conducted in two consecutive years (2017–2018 and 2018–2019) in the Fritissa-Mateur-Bizerte region. Our results revealed that barley considerably contributed to the decrease in G. coronaria’s density, cohort number, total biomass accumulation and plant height. The effect of rapeseed is likely dependent on the tillage practice and the timing of herbicide application, while G. coronaria could be successfully controlled in wheat using auxin herbicides. It is suggested that the management of resistant population of G. coronaria could be achieved by combining barely or rapeseed with right tillage practice and auxin herbicides’ application.

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Glebionis coronaria is a serious threat to cereal production in Northern Tunisia. Previous results showed that G. coronaria has developed resistance to acetolactate synthase (ALS)-inhibiting herbicides which limits their use in cereal cropping systems. The use of allelopathic crops has been reported as a potential alternative to herbicides to control resistant weed populations. The aim of this study is to evaluate the effect of five allelopathic crops on the early growth of G. coronaria. In vitro experiments using aqueous extracts of different crops and in pots experiments with increasing rates of barley (Hordeum vulgare) and rapeseed (Brassica napus) residues were performed for this purpose. All tested crops showed allelopathic effects on G. coronaria and were able to reduce its root length at the concentration of 50 g/L. Barley and rapeseed were the most effective crops, with 70% and 60% of root length reduction, respectively. An aqueous extract concentration study with five increasing concentrations of 12.5, 25, 50, 75, and 100 % was performed with these two crops, and root length was significantly reduced with increasing concentration, being up to 80% with the highest aqueous extract concentration of barley. The in vitro results were supported by the in pots experiment: Shortest roots length and highest dry mass reduction were obtained by the highest barley (0.64 g/kg) and rapeseed (8 g/kg) residue concentrations. This study confirmed the allelopathic potential of both barley and rapeseed in reducing the early growth of ALSresistant G. coronaria, suggesting the effectiveness of these crops if integrated with other control measures for the management of resistant population of G. coronaria in cereal fields.