The sustainability of rice cropping systems is jeopardized by the large number and variety of populations of polyploid Echinochloa spp. resistant to ALS inhibitors. Better knowledge of the Echinochloa species present in Italian rice fields and the study of ALS genes involved in target-site resistance could significantly contribute to a better understanding of resistance evolution and management. Using a CAPS-rbcL molecular marker, two species, E. crus-galli (L.) P. Beauv. and E. oryzicola (Vasinger) Vasing., were identified as the most common species in rice in Italy. Mutations involved in ALS inhibitor resistance in the different species were identified and associated with the ALS homoeologs. The relative expression of the ALS gene copies was evaluated. Molecular characterization led to the identification of three ALS genes in E. crus-galli and two in E. oryzicola. The two species also carried different point mutations conferring resistance: Ala122Asn in E. crus-galli and Trp574Leu in E. oryzicola. Mutations were carried in the same gene copy (ALS1), which was significantly more expressed than the other copies (ALS2 and ALS3) in both species. These results explain the high resistance level of these populations and why mutations in the other ALS copies are not involved in herbicide resistance.

Amaranthus palmeri S. Watson (Amaranthaceae Juss.) is a dioecious noxious weed, native to the Americas, which infests summer crops. It causes high crop losses, and rapidly evolves resistance to herbicides. In Europe, A. palmeri was recorded mostly as a casual alien, but in 2018 it was reported infesting a soybean field in Italy, and the next year two more populations were found in the same area. Experiments were conducted on these three populations to evaluate the resistance to ALS-inhibiting herbicides, to determine the main resistance mechanisms involved and assess the efficacy of alternative herbicides with different sites of action than ALS. The three populations were confirmed cross-resistant to ALS-inhibiting herbicides (thifensulfuron-methyl and imazamox). Gene sequencing identified a Trp to Leu substitution at position 574 of ALS gene in resistant plants, proving that the main resistance mechanism for the three populations is target-site related. The presence of other resistance mechanisms cannot be excluded. Metobromuron, metribuzin and glyphosate are still effective on these populations.

The invasive weed Amaranthus palmeri is spreading throughout Spain, with Catalonia being one of the most affected regions. For this species, acetolactate synthase‐inhibiting herbicide‐resistant populations have been reported, and now glyphosate resistance is also suspected. Glyphosate targets and inhibits the enzyme 5‐enolpyruvylshikimate‐3‐phosphate synthase (EPSPS), but A. palmeri has evolved different resistant mechanisms leading to plant survival. One of the most effective is the EPSPS overexpression due to copy number variation (CNV). Gene copies accumulate within the EPSPS cassette that is an extrachromosomal circular DNA displaying unique structural polymorphisms. This study aims to determine the response to glyphosate of a suspected resistant population collected from a roadside and investigate the resistance mechanism involved. The herbicide bioassay confirmed that 40% of the plants survived glyphosate applied at 540 g a.i. ha⁻¹. No known mutations endowing glyphosate resistance were found at EPSPS amongst confirmed resistant plants, while in most of them (70%) specific molecular markers revealed the presence of the EPSPS cassette. All these results indicate that this population is glyphosate resistant and it is very likely that the EPSPS gene CNV is the main resistance mechanism. This is the first case of glyphosate resistance in A. palmeri in Europe whose introduction is likely due to importation of contaminated seed with glyphosate‐resistant Palmer amaranth from the Americas. This introduction poses a significant danger to summer crops in our continent.