Herbicidal Effects of Sulfamethoxazole in Lemna gibba: Using p-Aminobenzoic Acid As a Biomarker of Effect

Sulfamethoxazole (SMX) is among the most frequently detected antibiotics in the environment, heavily used in both human therapy and agriculture. Like other sulfonamides, SMX disrupts the folate biosynthetic pathway in bacteria, which was recently established as identical to that of plants, raising concerns over nontarget toxicity. Consequently, Lemna gibba was exposed to SMX to evaluate phytotoxic potency and mode of action (MOA) by HPLC-MS/MS measurement of p-aminobenzoic acid (pABA) metabolite levels, a precursor to folate biosynthesis and substrate of the target enzyme dihydropteroate synthase (DHPS). pABA levels were found to increase upon exposure to SMX following an exponential rise to a maxima regression model in a concentration-dependent manner. The EC50 for pABA content was 3.36 microg/L, 20 times lower than that of fresh weight (61.6 microg/L) and 40 times lower than frond number (132 microg/L) responses. These results suggest that, as in bacteria, sulfonamide antibiotics specifically disrupt folate biosynthesis via inhibition of DHPS. Analysis of pABA concentrations appears to provide a sulfonamide-specific biomarker of effect based on MOA with exceptional diagnostic capacity and sensitivity compared to traditional morphological end points. Using the EC50 for pABA content, a potential hazard was identified for L. gibba exposed to SMX, which would not have been detected based upon traditional standardized morphological approaches.