Physiological and Molecular Responses of Woody Plants Exposed to Future Atmospheric CO2 Levels under Abiotic Stresses.

Climate change is mainly driven by the accumulation of carbon dioxide (CO2) in the atmosphere in the last century. Plant growth is constantly challenged by environmental fluctuations including heat waves, severe drought and salinity, along with ozone accumulation in the atmosphere. Food security is at risk in an increasing world population, and it is necessary to face the current and the expected effects of global warming. The effects of the predicted environment scenario of elevated CO2 concentration (e[CO2]) and more severe abiotic stresses have been scarcely investigated in woody plants, and an integrated view involving physiological, biochemical and molecular data is missing. This review highlights the effects of elevated CO2 in the metabolism of woody plants and the main findings of its interaction with abiotic stresses, including a molecular point of view, aiming to improve the understanding of how woody plants will face the predicted environmental conditions. Overall, e[CO2] stimulates photosynthesis and growth and attenuates mild to moderate abiotic stress in woody plants if root growth and nutrients are not limited. Moreover, e[CO2] does not induce acclimation in most tree species. Some high-throughput analyses involving omics techniques were conducted to better understand how these processes are regulated. Finally, knowledge gaps in the understanding of how the predicted climate condition will affect woody plant metabolism were identified, with the aim of improving the growth and production of this plant species.

Elevated CO2 Atmosphere Minimizes the Effect of Drought on the Cerrado Species Chrysolaena obovata.

Chrysolaena obovata stores inulin in the rhizophores, associated with drought tolerance. While crop plants are widely studied concerning the interactive effects of high [CO2] and drought, few studies reported these effects in native species. Here, we evaluated the combined effects of these factors on water status and fructan metabolism in C. obovata, a native Cerrado species. Two lots of plants were kept at 380 and 760 ppm CO2 in open-top chambers. In each, [CO2] plants were divided into four groups and cultivated under different water availability: irrigation with 100 (control), 75 (low), 50 (medium), and 25% (severe drought) of the water evapotranspirated in the last 48 h. In each, water treatment plants were collected at 0, 9, 18, and 27 days. On day 27, all plants were re-watered to field capacity and, after 5 days, a new sampling was made. Water restriction caused a decrease in plant moisture, photosynthesis, and in enzymes of fructan metabolism. These changes were generally more pronounced in 25% plants under ambient [CO2]. In the later, increases in the proportion of hexoses and consequent modification of the fructan chain sizes were more marked than under high [CO2]. The results indicate that under elevated [CO2], the negative effects of water restriction on physiological processes were minimized, including the maintenance of rhizophore water potential, increase in water use efficiency, maintenance of photosynthesis and fructan reserves for a longer period, conditions that shall favor the conservation of this species in the predicted climate change scenarios.

Structural and metabolic changes in rhizophores of the Cerrado species Chrysolaena obovata (Less.) Dematt. as influenced by drought and re-watering.

The high fructan contents in underground organs of Cerrado species, high water solubility, and fast metabolism of these compounds highlight their role as carbon storage and as an adaptive feature in plants under drought. In this study, we showed that anatomical structure, in association with soluble compounds and metabolism of inulin-type fructans were modified in rhizophores of Crysolaena obovata submitted to water suppression and recovery after re-watering. Plants were subjected to daily watering (control), suppression of watering for 22 days (water suppression) and suppression of watering followed by re-watering after 10 days (re-watered). Plants were collected at time 0 and after 3, 7, 10, 12, 17, and 22 days of treatment. In addition to changes in fructan metabolism, high proline content was detected in drought stressed plants, contributing to osmoregulation and recovery after water status reestablishment. Under water suppression, total inulin was reduced from approx. 60 to 40%, mainly due to exohydrolase activity. Concurrently, the activity of fructosyltransferases promoted the production of short chain inulin, which could contribute to the increase in osmotic potential. After re-watering, most parameters analyzed were similar to those of control plants, indicating the resumption of regular metabolism, after water absorption. Inulin sphero-crystals accumulated in parenchymatic cells of the cortex, vascular tissues and pith were reduced under drought and accompanied anatomical changes, starting from day 10. At 22 days of drought, the cortical and vascular tissues were collapsed, and inulin sphero-crystals and inulin content were reduced. The localization of inulin sphero-crystals in vascular tissues of C. obovata, as well as the decrease of total inulin and the increase in oligo:polysaccharide ratio in water stressed plants is consistent with the role of fructans in protecting plants against drought.

Leaf metabolite profile of the Brazilian resurrection plant Barbacenia purpurea Hook. (Velloziaceae) shows two time-dependent responses during desiccation and recovering.

Barbacenia purpurea is a resurrection species endemic to rock outcrops, in Rio de Janeiro, Brazil. It tolerates great temperature variations, which are associated to periods of up to 30 days without precipitation. Using a metabolomic approach, we analyzed, under winter and summer conditions, changes in the leaf metabolite profile (MP) of potted plants of B. purpurea submitted to daily watered and water deficit for at least 20 days and subsequent slow rehydration for 5 days. Leaves were collected at different time points and had their MP analyzed by GC/MS, HPAEC, and UHPLC techniques, allowing the identification of more than 60 different compounds, including organic and amino acids, sugars, and polyols, among others. In the winter experiment, results suggest the presence of two time-dependent responses in B. purpurea under water stress. The first one starts with the increase in the content of caffeoyl-quinic acids, substances with strong antioxidant activity, until the 16th day of water suppression. When RWC reached less than 80 and 70%, in winter and summer respectively, it was observed an increase in polyols and monosaccharides, followed by an increment in the content of RFO, suggesting osmotic adjustment. Amino acids, such as GABA and asparagine, also increased due to 16 days of water suppression. During rehydration, the levels of the mentioned compounds became similar to those found at the beginning of the experiment and when compared to daily watered plants. We conclude that the tolerance of B. purpurea to dehydration involves the perception of water deficit intensity, which seems to result in different strategies to overcome the gradient of water availability imposed along a certain period of stress mainly during winter. Data from summer experiment indicate that the metabolism of B. pupurea was already primed for drought stress. The accumulation of phenolics in summer seemed to be more temperature and irradiance-dependent than on the RWC.

Effect of drought and re-watering on fructan metabolism in Vernonia herbacea (Vell.) Rusby.

Vernonia herbacea (Vell.) Rusby, a native species from the Brazilian Cerrado, accumulates about 80% of fructans in the rhizophores, the underground reserve organs. Besides their role as reserve, fructans have been recognized as protective compounds against drought. This physiological function attributed to fructans seems consistent with the wide occurrence of these carbohydrates in the cerrado, a biome that undergoes seasonal drought. The aim of this work was to analyze fructan composition and the activities of the enzymes involved in fructan synthesis, sucrose:sucrose 1-frutosyltransferase (1-SST) and fructan:fructan 1-frutosyltransferase (1-FFT), and depolymerization, fructan 1-exohydrolase (1-FEH) in plants submitted to water suppression. The plants were divided into 3 groups receiving 3 treatments: daily watering (control), water suppression for 23 days (WS) and re-watering after 15 days (RW). Samples were taken at the beginning of the experiment (Time 0) and after 3, 7, 11, 15, 17 and 23 days of water suppression. 1-SST and 1-FFT activities increased at the beginning of the water restriction period, coinciding with a decrease in 1-FEH activity, the onset of the reduction in soil water potential and in leaf water potential. Increases in 1-FEH and invertase activities led to a high yield of reducing sugars at the 23rd day after water suppression, and together with 1-FFT, 1-FEH also seemed to act in the redistribution of fructan molecules after re-watering. The increase in reducing sugars and in the fructo-oligo:fructo-polysaccharide ratio were associated to the maintenance of rhizophore turgor. Considering that WS plants showed changes in fructan metabolism that favored water retention and absorption after re-watering, the occurrence of osmotic adjustment mechanisms is suggested, reinforcing the hypothesis of fructans as protective agents against abiotic stresses, such as drought.