Detection of ACCase target-site resistant Alopecurus myosuroides Huds. (black-grass) in Belgian populations

Black-grass is a common grass weed, widely spread in Northern Europe and also in Belgium. For ages, it has been an increasing problem in industrial crops, especially winter cereals. The first case of resistance in Belgium was reported in 1996 by Robert Bulcke (Eelen et al., 1996). Yet the resistance mechanism was not specified. Since then, no more information was published about the evolution Belgium, while research continued in the United Kingdom and in France. Moreover, during the last decade, progress in molecular biology allowed to highlight the mechanism of target-site resistance. A simple PCR method allows to detect the mutation conferring resistance to herbicide. After two years of resistance monitoring in Belgium, mostly in the Walloon part, some populations have been clearly identified as highly resistant to ACCase inhibitor. These populations have been tested by molecular biology so as to detect the single nucleotide polymorphism (SNP) involved in this case. The method employed was the Polymerase Chain Reaction Allele Specific Assays (PASA: Délye, 2002a) for the mutation Ile-1781-Leu that confers a target-site resistance to ACCase inhibitors. Those analyses were performed on plant material issued from bioassays, either in glasshouses or in Petri dishes. Leaves have been collected from plants which survived a fenoxaprop-P treatment applied in a glasshouse single dose assay. Seedlings from resistant populations grown in Petri dishes containing either fenoxaprop-P or cycloxydim provided the second type of sample. Ile1781 mutants were discovered within three populations. Each mutant plant was heterozygote. Five of those samples have been sequenced to confirm PASA results and everyone was matching. Moreover, they were all issued from Petri dishes containing cycloxydim, known to be unaffected by enhanced metabolism, confirming that theses populations are indeed target-site resistant.