BACKGROUND: Barnyardgrass (Echinochloa crus-galli (L.) P. Beauv) has evolved resistance to the acetolactate synthase (ALS) inhibitor penoxsulam which is used to control weeds in rice fields in China. The present study is conducted to identify the target-site resistance (TSR) mechanisms conferring resistance in a penoxsulam-resistant population.RESULTS: The ALS sensitivity in vitro of the resistant population was sixfold lower to penoxsulam than that of the sensitive population. ALS sequencing revealed that no known mutation conferring ALS herbicide resistance was detected. However, a novel mutation Phe-206-Leu was identified in the ALS gene. Additionally, ALS gene expression level of the resistant population was lower than that of the sensitive population. Therefore, the penoxsulam resistance was not due to the overexpression of ALS gene. Molecular docking revealed that this mutation may change the interaction of the penoxsulam-ALS binding and weaken its mutual affinity by approximately 10%. Arabidopsis thaliana transformed with mutant ALS had fourfold greater resistance to penoxsulam and varied cross-resistance to other ALS herbicides than those transformed with sensitive ALS. Mutant and sensitive ALS proteins expressed by the baculovirus system exhibited different in vitro penoxsulam sensitivity levels. Mutant ALS had eightfold lower sensitivity to penoxsulam than sensitive ALS.CONCLUSION: This report provides clear evidence that the ALS mutation at position 206 (Phe-206-Leu) confers penoxsulam resistance in barnyardgrass. Phe-206 was confirmed to be the ninth amino acid residue related to ALS herbicide resistance in weeds.
Differences in ethylene biosynthesis and cyanide detoxification have been reported to be mechanisms of quinclorac resistance in Echinochloa crusgalli var. zelayensis. Resistant phenotypes could be a consequence of the altered endogenous indole acetic acid (IAA) homeostasis induced by the herbicide. In this study, we determined the IAA content and expression levels of auxin homeostasis-related genes in susceptible and resistant biotypes of E. crusgalli var. zelayensis after quinclorac treatment. The results showed that the IAA content of JNNX-S (susceptible biotype) was significantly higher than that of SSXB-R (resistant biotype) after treatment with 50 μM quinclorac. To better understand this rise in IAA, the expression profiles of seven genes (one for auxin synthesis, five for IAA conjugation and one for IAA oxidation) and the biochemical activities of two oxidases involved in IAA homeostasis were measured. The expression of EcYUCCA10 was significantly higher in JNNX-S than in SSXB-R. The expression levels of the EcGH3s were significantly lower in JNNX-S than in SSXB-R. These expression profiles were consistent with the elevation of IAA levels in the susceptible biotype. In contrast, EcUGT and EcDAO were induced in each biotype, but a smaller increase was observed in SSXB-R than in JNNX-S. The enzymatic activities of IAA oxidases and peroxidases were higher in SSXB-R than in JNNX-S 24 h after treatment. It was inferred that altered expression of specific genes involved in IAA synthesis, conjugation and oxidation resulted in less IAA being induced in the resistant biotype, resulting in a lower ethylene burst and the associated quinclorac resistance. These results suggest novel layers of complexity in the mechanism of quinclorac resistance.
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