Photoprotective mechanisms and higher photorespiration are key points for iron stress tolerance under heatwaves in rice.

Considering the current climate change scenario, the development of heat-tolerant rice cultivars (Oryza sativa L.) is paramount for cultivation in waterlogged systems affected by iron (Fe) excess. The objective of this work was to investigate the physiological basis of tolerance to excess Fe in rice cultivars that would maintain photosynthetic efficiency at higher temperatures. In an experimental approach, two rice cultivars (IRGA424 - tolerant and IRGA417- susceptible to Fe toxicity) were exposed to two concentrations of FeSO4-EDTA, control (0.019 mM) and excess Fe (7 mM) and subsequent exposition to heatwaves at different temperatures (25 °C - control, 35, 40, 45, 50, and 55 °C). The increase in temperatures resulted in a higher Fe concentration in shoots accompanied by a lower Rubisco carboxylation rate in both cultivars, but with lower damage in the tolerant one. Stomatal limitation only occurred as a late response to Fe toxicity, especially in the sensitive cultivar. The activation of photorespiration as electron sink under Fe excess with increasing temperature during heatwaves appear as a major mechanism to alleviate oxidative stress in cultivars tolerant to excess Fe. The tolerance to iron toxicity and heat stress is associated with increased photoprotective mechanisms driving non-photochemical dissipation.

Photochemical attributes determine the responses of plant species from different functional groups of ferruginous outcrops when grown in iron mining substrates.

Environments originating from banded iron formations, such as the canga , are important reference ecosystems for the recovery of degraded areas by mining. The objective of this work was to evaluate if the relationship between morphofunctional and photosynthetic attributes of native canga species from different functional group results in distinct responses when grown in iron mining tailings substrate. The experiment was carried out with species belonging to different functional groups: a widespread semi-deciduous tree-shrub, Myrcia splendens ; an endemic deciduous shrub, Jacaranda caroba ; and a nitrogen-fixing herbaceous species, Periandra mediterranea . The species were grown in two conditions, reference soil and iron ore tailing. Despite belonging to different functional groups when grown in tailings, the morphofunctional attributes presented similar responses between species. M. splendens was the species most affected by the conditions imposed by the iron ore mining tailings, with decreased light-use efficiency and electron transport. P. mediterranea had satisfactory growth and maintenance of photosynthetic attributes. J. caroba growing in the tailings increased the effective quantum yield of PSII. The photochemical and growth assessments were able to better explain the adaptive strategies developed by the species, guaranteeing a greater chance of success during the rehabilitation of mining substrates.

Revegetation of an area impacted by iron ore tailings: evaluating fertilization alternatives in native pioneer and secondary trees.

The iron ore tailings released into the Rio Doce basin after the Fundão dam collapse (Brazil), suppressed a large extent of local vegetation. The use of native species and appropriate fertilization techniques, with less economic and environmental impact, must be considered in the process for the restoration of affected areas by the tailings. For this purpose, six native tree species, pioneer (Anadenanthera colubrina, Bixa orellana, and Peltophorum dubium) and secondary (Cedrela fissilis, Handroanthus impetiginosus, and Handroanthus serratifolius), were selected. We used different conditions of fertilization: (1) inorganic fertilization, (2) inoculation with arbuscular mycorrhizal fungi and plant growth-promoting rhizobacteria, (3) combined treatment (fertilizer + inoculum), to evaluate leaf nutrient concentrations, photosynthetic capacity [chlorophyll index, maximum quantum efficiency of photosystem II (Fv/Fm) and gas exchange variables], and oxidative metabolism (H2O2, MDA, and antioxidant enzymes). Inoculation resulted in higher concentrations of foliar nitrogen, especially in pioneer species. In all treatments, the secondary species exhibited iron values considered phytotoxic, but showed reduced photosynthetic capacity only when inoculated. The highest concentrations of MDA were observed in inoculated plants of both successional groups. The antioxidant system proved to be effective in preventing oxidative damage for most of the species. These results showed that the use of inoculum can be considered an ecological alternative to inorganic additives in the area affected by iron ore tailings. Despite presenting different photosynthetic and antioxidant strategies, the evaluated species demonstrated potential for use in tailings revegetation projects.

Revegetation of mining-impacted sites with a tropical native grass: Constraints of climate seasonality and trace-element accumulation.

The iron ore mining activity results in considerable waste production and impacts on surrounding ecosystems. Natural recovery of impacted areas is absent or occurs slowly, especially when associated with prolonged dry seasons in tropical regions. The objective of this work was to unveil the mechanisms of Paspalum densum (Poir.) grass to overcome the periods of seasonal drought and its metal accumulation in areas impacted by iron mining, a tailings storage facilities and surrounding ferruginous grassland in Brazil. Lower mortality was observed among individuals in the tailings storage facilities, with a 74.3% survival rate. In contrast, after the beginning of the dry season, all individuals died in the ferruginous grassland. The plants in the tailings deposits showed better nutrition, with a higher concentration of P, Cu, Fe, Zn, Mn and greater capacity to accumulate Pb and Cd over time. Pb was the element with highest bioconcetration factor (BCF) and bioaccumulation coefficient (BAC), while Mn was the one with the highest translocation factor (TF). The dry season resulted in reduced chlorophyll a, b and total and effective quantum yield of photosystem II (PSII) of P. densum individuals. However, the plants in the tailings storage facilities showed adjustments to overcome the effects of drought, with an increase in the concentration of proline in leaves and reduction of oxidative damage (MDA and H2O2) at the end of the dry season. The grass P. densum showed rapid acclimatization in the tailings storage facilities and resistance to drought through antioxidant and photosynthetic adjustments and was still able to bioaccumulate and translocate in plant tissues some metals present in the iron ore impacted sites.

Diversity of responses to nitrogen deficiency in distinct wheat genotypes reveals the role of alternative electron flows in photoprotection.

Nitrogen (N) deficiency represents an important limiting factor affecting photosynthetic productivity and the yields of crop plants. Significant reported differences in N use efficiency between the crop species and genotypes provide a good background for the studies of diversity of photosynthetic and photoprotective responses associated with nitrogen deficiency. Using distinct wheat (Triticum aestivum L.) genotypes with previously observed contrasting responses to nitrogen nutrition (cv. Enola and cv. Slomer), we performed advanced analyses of CO2 assimilation, PSII, and PSI photochemistry, also focusing on the heterogeneity of the stress responses in the different leaf levels. Our results confirmed the loss of photosynthetic capacity and enhanced more in lower positions. Non-stomatal limitation of photosynthesis was well reflected by the changes in PSII and PSI photochemistry, including the parameters derived from the fast-fluorescence kinetics. Low photosynthesis in N-deprived leaves, especially in lower positions, was associated with a significant decrease in the activity of alternative electron flows. The exception was the cyclic electron flow around PSI that was enhanced in most of the samples with a low photosynthetic rate. We observed significant genotype-specific responses. An old genotype Slomer with a lower CO2 assimilation rate demonstrated enhanced alternative electron flow and photorespiration capacity. In contrast, a modern, highly productive genotype Enola responded to decreased photosynthesis by a significant increase in nonphotochemical dissipation and cyclic electron flow. Our results illustrate the importance of alternative electron flows for eliminating the excitation pressure at the PSII acceptor side. The decrease in capacity of electron acceptors was balanced by the structural and functional changes of the components of the electron transport chain, leading to a decline of linear electron transport to prevent the overreduction of the PSI acceptor side and related photooxidative damage of photosynthetic structures in leaves exposed to nitrogen deficiency.

How does drought affect native grasses' photosynthesis on the revegetation of iron ore tailings?

The revegetation of areas degraded by iron ore mining is a difficult challenge mainly due to water availability and impoverished metal-rich substrates. We sought to understand the photosynthetic responses to drought of native tropical grasses Paspalum densum (Poir.) and Setaria parviflora (Poir.) grown in iron ore tailing. The grass P. densum presented better photosynthetic adjustments when grown in the iron ore tailing and S. paviflora in response to water stress. Both species accumulated iron above the phytotoxic threshold when grown in an iron ore tailing. The net photosynthesis, stomatal conductance, transpiration, and water use efficiency decreased followed by a reduction in leaf relative water content in response to water stress for both species. The photochemical efficiency of photosystem II only decreased at the point of maximum drought. At this point, the water-stressed grass grown in the iron ore tailing presented higher H2O2 concentrations, particularly S. parviflora. After rehydration, full recovery of photosynthetic variables was achieved with decreased malondialdehyde concentrations, increased catalase activity, and, consequently, decreased H2O2 concentrations in leaves for both species. The fast recovery of the native grasses P. densum and S. parviflora to drought in the iron ore tailing substrate is indicative of their resistance and potential use in the revegetation of impoverished mined areas with high iron content and seasonal water deficit.

Iron toxicity resistance strategies in tropical grasses: The role of apoplastic radicular barriers.

The revegetation of mined areas poses a great challenge to the iron ore mining industry. The initial recovery process in degraded areas might rely on the use of Fe-resistant grasses. Tropical grasses, such as Paspalum densum and Echinochloa crus-galli, show different resistance strategies to iron toxicity; however, these mechanisms are poorly understood. The Fe-resistance mechanisms and direct iron toxicity as a function of root apex removal were investigated. To achieve this purpose, both grass species were grown for up to 480 hr in a nutrient solution containing 0.019 or 7 mmol/L Fe-EDTA after the root apices had been removed or maintained. Cultivation in the presence of excess iron-induced leaf bronzing and the formation of iron plaque on the root surfaces of both grass species, but was more significant on those plants whose root apex had been removed. Iron accumulation was higher in the roots, but reached phytotoxic levels in the aerial parts as well. It did not hinder the biosynthesis of chloroplastidic pigments. No significant changes in gas exchange and chlorophyll a fluorescence occurred in either grass when their roots were kept intact; the contrary was true for plants with excised root apices. In both studied grasses, the root apoplastic barriers had an important function in the restriction of iron translocation from the root to the aerial plant parts, especially in E. crus-galli. Root apex removal negatively influenced the iron toxicity resistance mechanisms (tolerance in P. densum and avoidance in E. crus-galli).

Elevated CO2 concentration improves functional traits and early growth of the widespread species Enterolobium contortisiliquum (Vell. ) Morong / Elevada concentração de CO2 melhora as características funcionais e o crescimento inicial da espécie generalista Enterolobium contortisiliquum (Vell. ) Morong

Enterolobium contortisiliquum is a neotropical widespread species that occurs in native AtlanticForest, Cerrado and Caatinga vegetation in Brazil. Outside Brazil, it occurs in Argentina, Bolivia, Paraguay, and Uruguay. The objective of this study was to investigate the influence of high carbon dioxide concentration [CO2] on leaf gas exchange, chlorophyll content, chlorophyll a fluorescence, growth, and biomass allocation among roots, stems, and leaves in young plants of E. contortisiliquum. After germination, twenty days old individuals of E. contortisiliquum were grown in open-top chambers (OTC) at two different [CO2] (400 ppm and 700 ppm). Physiological measurements were made when the plants had been grown in OTC for 170, 200, 230, and 260 days. The higher [CO2] favored the early growth of E. contortisiliquum by increasing maximum net photosynthesis (PN) and water use efficiency. Under high [CO2], root dry mass increased in E. contortisiliquum, which might be a trait that will benefit this species by enhancing water uptake, particularly in seasonally dry environments.

Enterolobium contortisiliquum é uma espécie neotropical generalista, encontrada na vegetação nativa da Mata Atlântica do Brasil, no Cerrado e na Caatinga. Fora do país, também pode ser encontrada empaíses como Argentina, Bolívia, Paraguai e Uruguai. O objetivo deste estudo foi investigar a influência da alta concentração de dióxido de carbono [CO2] nas trocas gasosas foliares, no teor de clorofila, na fluorescência da clorofila a, no crescimento e alocação de biomassa entre raízes, caules e folhas em plantas jovens de E. contortisiliquum. Após a germinação, indivíduos de E. contortisiliquum de 20 dias de idade foram cultivados em câmaras de topo aberto (CTA), em duas diferentes [CO2] (400 ppm e 700 ppm). Asmedições fisiológicas foram realizadas no momento do cultivo das plantas em CTA por 170, 200, 230 e 260 dias. A alta [CO2] favoreceu o crescimento inicial de E. contortisiliquum aumentando a fotossíntese líquida máxima (PN) e a eficiência do uso da água. Sob alta [CO2], a massa seca da raiz aumentou em E. contortisiliquum, o que pode ser uma característica que beneficiará esta espécie, aumentando a absorção de água, particularmente em ambientes sazonalmente secos.

Physiological traits and antioxidant metabolism of leaves of tropical woody species challenged with cement dust.

Tropical woody species occurring in limestone outcrops are frequently exposed to particulate material from cement factories. The effects of 60-day cement dust exposure on physiological traits and enzymatic antioxidant system of young plant leaves of Guazuma ulmifolia Lam., Myracrodruon urundeuva Allemão and Trichilia hirta L. were investigated. Cement dust (2.5 or 5mgcm-2) was applied to the leaf surface or soil or both (leaf plus soil) and plants were maintained at greenhouse. Cement dust barely affected the mineral nutrient levels, except for iron whose content was decreased in leaves/leaflets of all species studied. The incident light was partly blocked in cement dust-treated leaves, regardless of the plant species, causing a decrease in the photosynthetic pigments in M. urundeuva. The chlorophyll b content, however, increased in G. ulmifolia and T. hirta leaves upon cement dust treatment. The potential quantum yield of photosystem II in challenged leaves of G. ulmifolia was 3.8% lower than that of control plants, while such trait remained unaffected in the leaves of the other species. No changes in leaf stomatal conductance and antioxidant enzymes activities were observed, except for M. urundeuva, which experienced a 31% increment in the superoxide dismutase activity upon 5mgcm-2 cement dust (leaf plus soil treatment), when compared with control plants. Overall, the mild changes caused by cement dust in the in physiological and biochemical traits of the species studied indicate that such species might be eligible for further studies of revegetation in fields impacted by cement factories.

Impact of cement dust pollution on Cedrela fissilis Vell. (Meliaceae): A potential bioindicator species.

Considering the impacts caused to vegetation in the vicinity of cement factories, the aim of this study was to evaluate the impacts of cement dust on the structural organization and physiological/biochemical traits of Cedrela fissilis leaflets, a woody species native to tropical America. Plants were exposed to 2.5 or 5 mg cm-2 cement dust applied to the leaf surface, to the soil or simultaneously to the leaf surface and the soil.. Leaves of shoot-treated plants exhibited chlorosis, marginal and inter veins necrosis, diminished thickness, epidermal cells less turgid, cellular collapse, obstructed stomata, senescence, rolling and some abscission. In few cases, individual death was recorded. Cement dust-treated plants also presented decreased amount of photosynthetic pigments and iron (Fe) and increase in calcium (Ca) levels. The cement crust formed in leaves surface blocked from 30 to 50% of the incoming light and reduced the stomatal conductance and the potential quantum yield of photosystem II. Control or soil-treated plants did not exhibit morphophysiological changes throughout the experiment. The activity of superoxide dismutase, catalase and ascorbate peroxidase increased in leaves of plants upon treatment with 2.5 mg cm(-2) cement dust, independent of the site application. Overall, these results indicate that C. fissilis is highly sensitive to cement dust at the initial stage of development.

Leaf structural traits of tropical woody species resistant to cement dust.

Cement industries located nearby limestone outcrops in Brazil have contributed to the coating of cement dust over native plant species. However, little is known about the extent of the response of tropical woody plants to such environmental pollutant particularly during the first stages of plant development and establishment. This work focused on the investigation of possible alterations in leaf structural and ultrastructural traits of 5-month-old Guazuma ulmifolia Lam. (Malvaceae), 6-month-old Myracrodruon urundeuva Allemão (Anacardiaceae), and 9-month-old Trichilia hirta L. (Meliaceae) challenged superficially with cement dust during new leaf development. Leaf surface of plants, the soil or both (leaf plus soil), were treated (or not) for 60 days, under controlled conditions, with cement dust at 2.5 or 5.0 mg cm(-2). After exposure, no significant structural changes were observed in plant leaves. Also, no plant death was recorded by the end of the experiment. There was also some evidence of localized leaf necrosis in G. ulmifolia and T. hirta, leaf curling in M. urundeuva and T. hirta, and bulges formation on epidermal surface of T. hirta, after cement dust contact with plant shoots. All species studied exhibited stomata obliteration while T. hirta, in particular, presented early leaf abscission, changes in cellular relief, and organization and content of midrib cells. No significant ultrastructural alterations were detected under the experimental conditions studied. Indeed, mesophyll cells presented plastids with intact membrane systems. The high plant survival rates, together with mild morphoanatomic traits alterations in leaves, indicate that G. ulmifolia is more resistant to cement dust pollutant, followed by M. urundeuva and T. hirta. Thus, the three plant species are promising for being used to revegetate areas impacted by cement industries activities.

Morphoanatomical responses induced by excess iron in roots of two tolerant grass species.

We aimed to verify whether morphoanatomic alterations occur in response to excess iron, in roots of Setaria parviflora and Paspallum urvillei (Poaceae), and to localize the presence of the sites of iron accumulation. Plants were subjected to 0.009, 1, 2, 4, and 7 mM Fe-EDTA in nutrient solution. Both species presented iron contents in the roots above the critical toxicity level. The presence of iron plaque on roots of the two species was confirmed, and it may have reduced iron absorption by the plants. Roots from the two species showed typical visual symptoms of stress by excess iron: change in color and mucilaginous and flaccid appearance. Anatomical damage was observed in both species: aerenchyma disruption, alterations in endodermal cells, and irregular shape of both vessel and sieve tube elements. The metal was histolocalized in the cortex and in protoxylem and metaxylem cell walls in both species, which suggests a detoxification strategy for the excess iron. Phenolic compounds were not histolocalized in roots. Microscopic analyses were therefore effective in evaluating the real damage caused by excess iron.

Tolerance to iron accumulation and its effects on mineral composition and growth of two grass species.

This study aimed to assess the influence of excess iron on the capacity of accumulation of this heavy metal, mineral composition, and growth of Setaria parviflora and Paspalum urvillei. Seedlings were submitted to 0.009; 1; 2; 4; and 7 mM of Fe-EDTA. In both species there was an increase in the concentration of Fe, Zn, P, and Ca and a decrease in Mn, K, and Mg in the iron plaque. Both species accumulated more iron in roots. In the shoots, S. parviflora showed higher iron content, except at 7 mM. Iron altered the contents of Fe, Cu, K, and Mg in roots, and of Fe, Mn, Zn, N, P, K, Ca, and Mg in shoots. The two species tolerated high iron concentrations and accumulated high content of this element in both shoots and roots. The iron did not reduce their growth. Both species are indicated for studies aiming restoration of iron-contaminated areas.

Leaf morphoanatomy of species tolerant to excess iron and evaluation of their phytoextraction potential.

Setaria parviflora (Poir.) Kerguélen and Paspalum urvillei Steudel are grasses that grow naturally in a soil with high iron contents. This study aimed to characterize morphoanatomically and histochemically the iron phytotoxicity on leaves and evaluate the phytoextraction potential of these grasses. Saplings were cultivated in hydroponic solution with and without excess Fe-EDTA. Regarding measurements taken on leaves, reduction was observed among treatments of Fe-EDTA on height values of abaxial epidermis and bundle sheath in both species. As for iron histolocalization, stronger reaction was observed in leaves of S. parviflora, in comparison with P. urvillei. Anatomical damage, such as protoplast retraction, irregular xylem, changes in cell volume, and cell collapse, and visual symptoms, like leaf bronzing, chlorosis, and necrosis, were similar in both species when exposed to excess iron; however, P. urvillei showed more severe damage. This species accumulated more iron in shoots than S. parviflora and therefore is more favorable for use in phytoextraction. The root system of both species accumulated higher iron concentrations in relation to shoots.

Iron excess affects rice photosynthesis through stomatal and non-stomatal limitations.

Iron toxicity is the most important stressor of rice in many lowland environments worldwide. Rice cultivars differ widely in their ability to tolerate excess iron. A physiological evaluation of iron toxicity in rice was performed using non-invasive photosynthesis, photorespiration and chlorophyll a fluorescence imaging measurements and chlorophyll content determination by SPAD. Four rice cultivars (BR IRGA 409; BR IRGA 412; BRA 041171 and BRA 041152) from the Brazilian breeding programs were used. Fe(2+) was supplied in the nutrient solution as Fe-EDTA (0.019, 4, 7 and 9 mM). Increases in shoot iron content due to Fe(2+) treatments led to changes in most of the non-invasive physiological variables assessed. The reduction in rice photosynthesis can be attributed to stomatal limitations at moderate Fe(2+) doses (4mM) and both stomatal and non-stomatal limitations at higher doses. Photorespiration was an important sink for electrons in rice cultivars under iron excess. A decreased chlorophyll content and limited photochemical ability to cope with light excess were characteristic of the more sensitive and iron accumulator cultivars (BRA 041171 and BRA 041152). Chlorophyll fluorescence imaging revealed a spatial heterogeneity of photosynthesis under excessive iron concentrations. The results showed the usefulness of non-invasive physiological measurements to assess differences among cultivars. The contributions toward understanding the rice photosynthetic response to toxic levels of iron in the nutrient solution are also discussed.

Photosynthesis and oxidative stress in the restinga plant species Eugenia uniflora L. exposed to simulated acid rain and iron ore dust deposition: potential use in environmental risk assessment.

The Brazilian sandy coastal plain named restinga is frequently subjected to particulate and gaseous emissions from iron ore factories. These gases may come into contact with atmospheric moisture and produce acid rain. The effects of the acid rain on vegetation, combined with iron excess in the soil, can lead to the disappearance of sensitive species and decrease restinga biodiversity. The effects of iron ore dust deposition and simulated acid rain on photosynthesis and on antioxidant enzymes were investigated in Eugenia uniflora, a representative shrub species of the restinga. This study aimed to determine the possible utility of this species in environmental risk assessment. After the application of iron ore dust as iron solid particulate matter (SPM(Fe)) and simulated acid rain (pH 3.1), the 18-month old plants displayed brown spots and necrosis, typical symptoms of iron toxicity and injuries caused by acid rain, respectively. The acidity of the rain intensified leaf iron accumulation, which reached phytotoxic levels, mainly in plants exposed to iron ore dust. These plants showed the lowest values for net photosynthesis, stomatal conductance, transpiration, chlorophyll a content and electron transport rate through photosystem II (PSII). Catalase and superoxide dismutase activities were decreased by simulated acid rain. Peroxidase activity and membrane injury increased following exposure to acid rain and simultaneous SPM(Fe) application. Eugenia uniflora exhibited impaired photosynthetic and antioxidative metabolism in response to combined iron and acid rain stresses. This species could become a valuable tool in environmental risk assessment in restinga areas near iron ore pelletizing factories. Non-invasive evaluations of visual injuries, photosynthesis and chlorophyll a fluorescence, as well as invasive biochemical analysis could be used as markers.

Effects of simulated deposition of acid mist and iron ore particulate matter on photosynthesis and the generation of oxidative stress in Schinus terebinthifolius Radii and Sophora tomentosa L.

Particulate matter is a natural occurrence in the environment, but some industries, such as the iron ore sector, can raise the total amount of particles in the atmosphere. This industry is primarily a source of iron and sulfur dioxide particulates. The effects of the pollutants from the iron ore industries on representatives of restinga vegetation in a Brazilian coastal ecosystem were investigated using physiological and biochemical measures. Two species, Schinus terebinthifolius and Sophora tomentosa, were exposed to simulated deposition of acid mist and iron ore particulate matter in acrylic chambers in a greenhouse. Parameters such as gas exchange, fluorescence emission, chlorophyll content, total iron content, antioxidant enzyme activity and malondialdehyde content were assessed in order to evaluate the responses of the two species. Neither treatment was capable of inducing oxidative stress in S. terebinthifolius. Nevertheless, the deposition of iron ore particulates on this species increased chlorophyll content, the maximum quantum efficiency of photosystem II and the electron transport rate, while iron content was unaltered. On the other hand, S. tomentosa showed a greater sensitivity to the treatments. Plants of S. tomentosa that were exposed to acid mist had a decrease in photosynthesis, while the deposition of iron particulate matter led to an increase in iron content and membrane permeability of the leaves. The activities of antioxidant enzymes, such as catalases and superoxide dismutase, were enhanced by both treatments. The results suggested that the two restinga species use different strategies to overcome the stressful conditions created by the deposition of particulate matter, either solid or wet. It seems that while S. terebinthifolius avoided stress, S. tomentosa used antioxidant enzyme systems to partially neutralize oxidative stress. The findings also point to the potential use of S. tomentosa as a biomarker species under field conditions.