Descurainia sophia is one of the most notorious broadleaf weeds in China and has evolved extremely high resistance to acetolactate synthase (ALS)-inhibiting herbicide tribenuron-methyl. The target-site resistance due to ALS gene mutations was well-known, while the non-target-site resistance is not yet well-characterized. Metabolic resistance, which is conferred by enhanced rates of herbicide metabolism, is the most important NTSR. To explore the mechanism of metabolic resistance underlying resistant (R) D. sophia plants, tribenuron-methyl uptake and metabolism levels, qPCR reference gene stability, and candidate P450 genes expression patterns were investigated. The results of liquid chromatography-mass spectrometry (LC-MS) analysis indicated that the metabolic rates of tribenuron-methyl in R plants was significantly faster than in susceptible (S) plants, and this metabolism differences can be eliminated by P450 inhibitor malathion. The genes for 18S rRNA and TIP41-like were identified as the most suitable reference genes using programs of BestKeeper, NormFinder, and geNorm. The P450 gene CYP96A146 constitutively overexpressed in R plants compared to S plants; this overexpression in R plants can be suppressed by malathion. Taken together, a higher expression level of P450 genes, leading to higher tribenuron-methyl metabolism, appears to be responsible for metabolic resistance to tribenuron-methyl in R D. sophia plants.
Keywords: CYP96A146; Descurainia sophia L.; cytochrome P450 monooxygenases; gene expression; metabolic resistance; tribenuron-methyl.