The extensive use of pesticides combined with their persistence in the environment requires new methodologies to assess more effectively the population exposure to pesticides via air pollution. Biomonitoring pesticides with lichens has been poorly documented, although it represents a complementary approach to the usual active samplings, with an exposure to pesticides accumulated and integrated over several months. An optimized extraction procedure from the lichen Xanthoria parietina followed by a gas chromatographic-tandem mass spectrometric analysis is proposed here to quantify simultaneously 48 pesticides considered in France as priority active substances to monitor in the air. This method has been applied to lichen samples collected in 24 sites in southern France covering urban, industrial, and agricultural areas in order to identify potential contrasts related to anthropogenic activities. Fifteen pesticides (six fungicides, five insecticides, and four herbicides), including four active compounds currently banned by EU legislation, were detected in at least one site. Lindane, diflufenican, difenoconazole, and boscalid were the most common pesticides found in all sites. Urban sites appeared generally less contaminated compared to industrial and rural ones, but a strong heterogeneity was noticed between locations. The biomonitoring with lichens revealed unexpected contaminated areas, partly due to the use of herbicides for vegetation control in industrial and railway installations. The spatial distribution also suggests an input of pesticides by atmospheric transport at the local and regional scales.