Caractérisation de la résistance non liée à la cible aux herbicides inhibiteurs de l'ACCase et de l'ALS chez le vulpin des champs (Alopecurus myosuroides)

Abstract : Herbicides inhibiting Acetyl-CoenzymeA carboxilase (ACCase) and Acetohydroxyacid synthase (AHAS) are among the most widely used in France and in the world. Their intensive use selected resistant plants into weed populations. It was assessed that non-target-site resistance (NTSR) to herbicides inhibiting ACCase is prevalent on target-site resistance (TSR) in many black grass (Alopecurus myosuroides) populations which is a very harmful weed in the cereal fields. While TSR mechanisms are well know and understood, NTSR mechanisms have not been studied much by now. We tried to identify genes conferring NTSR to ACCase inhibitors and AHAS inhibitors into black grass thanks to a Suppressive Subtractive Hybridization (SSH) technology of analysis of the transcriptome. Characterization of NTSR involves a five steps process: to look for populations containing NTSR individuals, to characterize the phenotype of each individual of these populations, to set resistant and sensitive plants apart before and after the herbicide treatment into a freezer, to identify NTSR "candidate genes", and check the role of these genes in NTSR mechanisms. This work is about the first four steps of the process, which have been done with herbal material from different plants and populations because of the short time available. NTSR is an unforeseeable adaptive trait involving a large set of genes, and influenced by external factors. By characterizing the phenotype of many plants with several herbicides inhibiting ACCase and AHAS, we could assess that NTSR is a complex, polygenic and unforeseeable process. It relies on the accumulation of genes, each conferring a different level of resistance to each herbicide. An aim of this work is to identify some of these genes and to understand how they get involved in NTSR. We noticed that NTSR genes selected with herbicides inhibiting ACCase also encodes enzymes involved in NTSR to herbicides inhibiting AHAS. Different levels of expression or different sequences of NTSR genes both confer a level of resistance to the plants. Several biochemical functions seem to be involved, and two steps can be focused on: the perception of the herbicide and the transduction of the signal (regulators, kinases proteins), and the action of effectors (transport, response to stress, detoxification proteins). We could identify four NTSR "candidate genes": two of them are regulators, the other ones are effectors. A glycosyl transferase protein (GT) seems to be over expressed has a constitutive gene in resistant plants compared to sensitive plants, and a protein probably involved in response to oxidative stress is over expressed in resistant plants both before the herbicide treatment and as a result of the herbicide perception. We also identified many proteins putatively involved in a response to oxidative and environmental stresses.
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Master thesis
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http://docnum.univ-lorraine.fr/public/SCDSCI_M_2011_DUHOUX_ARNAUD.pdf
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Arnaud Duhoux. Caractérisation de la résistance non liée à la cible aux herbicides inhibiteurs de l'ACCase et de l'ALS chez le vulpin des champs (Alopecurus myosuroides). Sciences de l'environnement. 2011. ⟨hal-01877351⟩

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