Ozone reactivity and free radical scavenging behavior of phenolic secondary metabolites in lichens exposed to chronic oxidant air pollution from Mexico City

Lichen secondary metabolites putatively protect lichens from a variety of environmental stress factors, but it is unknown whether these substances respond to air pollution. To assess such a possibility, the three major phenolics of two epiphytic lichen species with contrasting tolerance to chronic air pollution from Mexico City were studied by combining experimental reactivity data and measured field contents. The antioxidant activity and antiradical power of boninic ( BO), 2- O- methylsekikaic ( MA), and usnic ( US) acids, isolated from the tolerant Ramalina asahinae and salazinic acid ( SA), atranorin ( AT), and chloroatranorin ( CA), from the sensitive Parmotrema stuppeum, were determined in vitro by kinetic experiments with ozone and the free radical diphenyl picryl hidrazyl ( DPPH center dot), respectively. In addition, the field contents of these phenolics in the lichens, and the potential antioxidant capacity ( PAC) they provide, were compared among three forested sites exposed to urban emissions and a similar, relatively clean site. The six phenolics had antioxidant activity and antiradical power according to these trends: CA>> AT> US> SA >= BO >= MA for O-3; and CA> AT> US> MA> SA= BO for DPPH center dot. The three most reactive phenolics are cortical compounds, located in the lichen portion most exposed to the surrounding environment. In contrast, the less reactive SA, BO, and MA are medullary. Such reactivity patterns indicate that some phenolics may provide antioxidative protection at the air - lichen interface. The higher antioxidant power of CA and AT may be due to the reactive hydroxyl groups at positions 2 and 4 of ring A, instead of the less reactive methoxyl at the same positions in both BO and MA. In the field comparisons, total quantified phenolics were significantly higher near Mexico City for both lichens, except for the tolerant R. asahinae at one site. Nevertheless, only the latter species had significantly increased PAC values at all sites near the city. This result is explained by species-dependent changes in individual phenolics. At the polluted sites, R. asahinae had consistently higher contents of its most reactive phenolic, US, with values approximately twice that of the control site. In contrast, P. stuppeum only increased its less reactive SA ( 26 - 35%), but this was counteracted by CA and, to a lesser extent, AT degradation. Thus, the substantial increase in US at the polluted sites appears to be associated with the current ecological success of R. asahinae near the city. On the other hand, the inability of P. stuppeum to overcome degradation of its most reactive phenolic (CA) at the same sites seems to partially explain the declining status of this lichen. These results provide evidence for a protective mechanism in lichens against air pollution based on secondary metabolites, which may eventually determine which species survive in forests stressed by oxidative air pollution.