Metabolic and physiological alterations indicate that the tropical broadleaf tree Eugenia uniflora L. is sensitive to ozone.

Eugenia uniflora L. is an important fruit tree native to tropical South America that adapts to different habitats, thanks to its metabolic diversity and ability to adjust the leaf antioxidant metabolism. We hypothesized that this metabolic diversity would also enable E. uniflora to avoid oxidative damage and tolerate the enhanced ozone (O3) concentrations that have been registered in the (sub)tropics. We investigated whether carbohydrates, polyphenols and antioxidants are altered and markers of oxidative damage (ROS accumulation, alterations in leaf gas exchange, growth and biomass production) are detected in plants exposed to two levels of O3 (ambient air and twice elevated ozone level in a O3-FACE system for 75 days). Phytotoxic O3 dose above a threshold of 0 nmol m-2 s-1 (POD0) and accumulated exposure above 40 ppb (AOT40) were 3.6 mmol m-2 and 14.898 ppb h at ambient, and 4.7 mmol m-2 and 43.881 ppb h at elevated O3. Twenty-seven primary metabolites and 16 phenolic compounds were detected in the leaves. Contrary to the proposed hypothesis that tropical broadleaf trees are relatively O3 tolerant, we concluded that E. uniflora plants are sensitive to elevated O3 concentrations. Experimental POD0 values were lower than the critical levels for visible foliar O3, because of low stomatal conductance. In spite of this low stomatal O3 uptake, we found classic O3 injury, e.g. reduction in carbohydrates and fatty acids concentrations; non-significant changes in the polyphenol profile; inefficient antioxidant responses; increased contents of ROS and indicators of lipid peroxidation; reductions in stomatal conductance, net photosynthesis, root/shoot ratio and height growth. However, we also found some compensation mechanisms, e.g. increased leaf concentration of polyols for protecting the membranes, and increased leaf number for compensating the decline of photosynthetic rate. These results help filling the knowledge gap about tropical tree responses to O3.

Does Phoradendron perrottetii (mistletoe) alter polyphenols levels of Tapirira guianensis (host plant)?

The present study aimed to investigate the reciprocal effects of Phoradendron perrottetii (mistletoe) and T. guianensis (host plant) regarding their polyphenol composition. Taking into account that tannins are important molecules in plant defense and their biosynthesis tends to be enhanced when a species is exposed to stress, we address the following questions: (1) Are the tannins found in our model species important in the interaction between host and mistletoe? (2) Does the presence of mistletoe induce changes in the content of tannins and other polyphenols in the host plant? (3) Do we find differences between the tannin sub-groups in the responses of the host plant to mistletoe? (4) Could the observed differences reflect the relative importance of one tannin group over another as chemical defense against the mistletoe? Using a polyphenol and tannin group-specific MRM methods we quantified four different tannin sub-groups together with flavonoid and quinic acid derivatives by ultra-performance liquid chromatography tandem mass spectrometry together with the oxidative and protein precipitation activities of leaves and branches of Tapirira guianensis and Phoradendron perrottetii. We selected leaves and branches of six non-parasitized trees of T. guianensis. Leaves and branches of nine individuals of T. guianensis parasitized by P. perrottetii were also sampled. For each parasitized tree, we sampled an infested branch and its leaves, as well as a non-infested branch and its leaves. Infested branches were divided into three groups: gall (the host-parasite interface), proximal, and distal region. Both proanthocyanidins and ellagitanins seem to be important for plant-plant parasitism interaction: host infested tissues (gall and surrounding regions) have clearly less tannin contents than healthy tissues. Mistletoe showed high levels of quinic acid derivatives and flavonoids that could be important during hastorium formation and intrusion on host tissues, suggesting a defense mechanism that could promote oxidative stress together with an inhibition of mistletoe seed germination, consequently avoiding secondary infestations. Polyphenol detected in T. guianensis-P. perrottetii interaction could play different role as plant-mistletoe strategies of survival.

The passion fruit liana (Passiflora edulis Sims, Passifloraceae) is tolerant to ozone.

Passiflora edulis Sims is a liana species of high economic interest and is an interesting model plant for understanding ozone action on disturbed vegetation. In this work we hypothesized that P. edulis has adaptive responses to oxidative stress that enable it to tolerate ozone damage based on its capacity to grow under a diversity of environmental conditions and to dominate disturbed areas. We exposed seedlings to three levels of ozone in a Free-Air Controlled Exposure (FACE) system (22, 41 and 58 ppb h AOT40 and 13.52, 17.24 and 20.62 mmol m-2 POD0, over 97 days) for identifying its tolerance mechanisms. Anatomical (leaf blade structure and fluorescence emission of chloroplast metabolites), physiological (leaf gas exchange, growth rate and biomass production) and biochemical (pigments, total sugars, starch, enzymatic and non-enzymatic antioxidant metabolites, reactive oxygen species and lipid peroxidation derivatives) responses were assessed. Ozone caused decreased total number of leaves, hyperplasia and hypertrophy of the mesophyll cells, and accelerated leaf senescence. However, O3 did not affect carbohydrates content, net photosynthetic rate, or total biomass production, indicating that the carboxylation efficiency and associated physiological processes were not affected. In addition, P. edulis showed higher leaf contents of ascorbic acid, glutathione (as well high ratio between their reduced and total forms), carotenoids, and flavonoids located in the chloroplast outer envelope membrane. Our results indicate that P. edulis is an O3-tolerant species due to morphological acclimation responses and an effective antioxidant defense system represented by non-enzymatic antioxidants, which maintained the cellular redox balance under ozone.