The damage caused by Cd toxicity to photosynthesis, cellular ultrastructure, antioxidant metabolism, and gene expression in young cacao plants are mitigated by high Mn doses in soil.

Manganese (Mn) is one of the essential mineral micronutrients most demanded by cacao. Cadmium (Cd) is highly toxic to plants and other living beings. There are indications that Mn can interact with Cd and mitigate its toxicity. The objective of this study was to evaluate the action of Mn on the toxic effect of Cd in young plants of the CCN 51 cacao genotype, subjected to different doses of Mn, Cd, and Mn+Cd in soil, through physiological, biochemical, molecular, and micromorphological and ultrastructural changes. High soil Mn doses favored the maintenance and performance of adequate photosynthetic processes in cacao. However, high doses of Cd and Mn+Cd in soil promoted damage to photosynthesis, alterations in oxidative metabolism, and the uptake, transport, and accumulation of Cd in roots and leaves. In addition, high Cd concentrations in roots and leaf tissues caused irreversible damage to the cell ultrastructure, compromising cell function and leading to programmed cell death. However, there was a mitigation of Cd toxicity when cacao was grown in soils with low Cd doses and in the presence of Mn. Thus, damage to the root and leaf tissues of cacao caused by Cd uptake from contaminated soils can be attenuated or mitigated by the presence of high Mn doses in soil.

Uranium transfer in grasses grown on mining waste and natural soil.

The transfer of radionuclides from soil to the food chain often begins with uptake by plant root system. The roots of most angiosperms showed symbiosis with arbuscular mycorrhizal fungi (AMF) and to understand the transfer process of these toxic elements it is important to consider different physical, chemical and biological factors in soils. In the present study, three grass species (Poaceae), Zea mays, Chrysopogon zizanioides and Aristida setifolia were cultivated with and without organic fertilization in experimental blocks on natural soils, at Fazenda Vargem Formosa (VF) with low uranium (U) contents in the soil, and in the leached ore deposit at the Uranium Concentrate Unit Mine (URA) in Caetité (Uraniferous Province of Lagoa Real - Brazil). In the present study, the biomass production of plants, their rate of root colonization by AMF, the levels of U in soils, roots and leaves, as well as different physico-chemical parameters related to soil fertility were evaluated. The data analysis was performed using Artificial Neural Networks (ANNs), specifically Self-Organizing Maps (SOMs). The levels of available uranium in the soil ranges from 0.33 to 1.11 mg kg-1 in VF and from 177.5 to 475.8 mg kg-1 in URA. The results revealed high percentage of root AMF colonization, even in soils with high U contents. There was an inverse relationship between soil U content and its transfer to the plant organs, with U transfer rates being influenced by plant species and not by soil parameters. C. zizanioides had the lowest transfer factor to the shoot and the highest mass productivity under conditions of high U content in soil. The results indicate that C. zizanioides is an important species for use in the recovery of U mining areas.

Witches' broom resistant genotype CCN51 shows greater diversity of symbiont bacteria in its phylloplane than susceptible genotype catongo.

BACKGROUND: Theobroma cacao L. (cacao) is a perennial tropical tree, endemic to rainforests of the Amazon Basin. Large populations of bacteria live on leaf surfaces and these phylloplane microorganisms can have important effects on plant health. In recent years, the advent of high-throughput sequencing techniques has greatly facilitated studies of the phylloplane microbiome. In this study, we characterized the bacterial microbiome of the phylloplane of the catongo genotype (susceptible to witch's broom) and CCN51 (resistant). Bacterial microbiome was determined by sequencing the V3-V4 region of the bacterial 16S rRNA gene. RESULTS: After the pre-processing, a total of 1.7 million reads were considered. In total, 106 genera of bacteria were characterized. Proteobacteria was the predominant phylum in both genotypes. The exclusive genera of Catongo showed activity in the protection against UV radiation and in the transport of substrates. CCN51 presented genus that act in the biological control and inhibition in several taxonomic groups. Genotype CCN51 presented greater diversity of microorganisms in comparison to the Catongo genotype and the total community was different between both. Scanning electron microscopy analysis of leaves revealed that on the phylloplane, many bacterial occur in large aggregates in several regions of the surface and isolated nearby to the stomata. CONCLUSIONS: We describe for the first time the phylloplane bacterial communities of T. cacao. The Genotype CCN51, resistant to the witch's broom, has a greater diversity of bacterial microbioma in comparison to Catongo and a greater amount of exclusive microorganisms in the phylloplane with antagonistic action against phytopathogens.

Mechanical stress caused by wind on leaves of Theobroma cacao: Photosynthetic, molecular, antioxidative and ultrastructural responses.

Theobroma cacao is cultivated in the shade, in a so-called 'Cabruca' system, in intercropped with Erithryna or other tree species of economic value, and in full sun as a monoculture in irrigated or chemically-irrigated systems. Since it is a species quite intolerant to wind, it is practically impossible to implant cacao crops under full exposure to the sun, or in areas of frequent winds, without the protection of windbreaks, using arboreal species around the area of culture in the form of box. Wind can cause mechanical stimuli in plants, affecting their growth and development. The objective of this work was to evaluate the photosynthetic changes in mature leaves and the molecular, biochemical and ultrastructural changes in young and mature leaves of the CCN 51 cloned genotype of T. cacao subjected to intermittent (IW) and constant (CW) wind, with velocities of 2.5, 3.5 and 4.5 m s-1, during 3, 6 and 12 h of exposure. It was verified that CW and IW, considering different exposure times, interfered directly in stomatal conductance (gs), transpiration (E) and water use efficiency (WUE), causing a reduction of the photosynthetic rate (A) in mature leaves. In addition, the pulvinus and blade of young and mature leaves, exposed to IW and CW with different exposure times (3 and 12 h), showed marked macroscopic and microscopic mechanical injuries resulting from the constant leaf movement. At both speeds, there was rupture of the cell nuclear membrane in pulvinus and the mesophyll tissues, mainly in the young leaves. On the other hand, in young and mature leaves exposed to CW and IW at different speeds and exposure times, there was lipid peroxidation, increased activity of guaiacol (GPX) and ascorbate (APX) peroxidases in most treatments; and altered expression of transcripts of psba and psbo genes related to the phothosynthetic apparatus and Cu-Zn-sod and per genes related to antioxidative enzymes at the rate of 4.5 m s-1. Younger leaves were more intolerant to mechanical stress caused by the wind, since presented greater macro and microscopic damages and, consequently, greater molecular, biochemical and ultrastructural changes. High wind speeds can seriously compromise the development of young leaves of T. cacao plants and affect their productivity.

Morphology, ultrastructure, and element uptake in Calophyllum brasiliense Cambess. (Calophyllaceae J. Agardh) seedlings under cadmium exposure.

Cadmium (Cd) is a metal known for its genotoxicity and cytotoxicity, much concerned for its potential environmental and human health impacts. This study evaluates the toxic effect of Cd in Calophyllum brasiliense plants. The plants were cultivated for 30 days in full nutrient solution in order to adapt, and for 15 days in nutrient solution without Cd or with 4, 8, 16, and 32 µmol Cd L-1. Anatomical analysis of the leaf showed no significant effects of Cd on epidermal thickness in abaxial and adaxial sides, palisade, and spongy parenchyma. Contrastingly, changes were noticed in the ultrastructural level in the leaf mesophyll cells as rupture of the membrane of chloroplasts and disorganization of the thylakoid membranes, in starch grains and in mitochondria with rupture of the membrane and invagination of the nuclear membrane. Electron dense materials into cells of the cortex and vascular bundle were also observed. In the cells of the root system, the observed ultrastructural changes were disruption of the cell wall and electron dense material deposition in the cortex cells and vascular region. Cd accumulated in roots with low translocation into shoot. Cd toxicity also affected the photosynthetic activity, inducing stomatal closure and photosynthetic assimilation reduction and the instantaneous carboxylation efficiency, drastically reducing the leaf transpiration. The nutrient content in the stem and root was variable, according to Cd increase in nutrient solution. Based on the experimental evidence, it can be concluded that C. brasiliense has potential to bioconcentrate high Cd levels in the root system.

Different salt concentrations induce alterations both in photosynthetic parameters and salt gland activity in leaves of the mangrove Avicennia schaueriana.

Mangrove sites are constantly under tidal regimes, where there is variation of dissolved oxygen and saline content in water. This study evaluated the effects of varying salinity in Avicennia schaueriana, an abundant species in the sea-mangrove-river interface. Seedlings of A. schaueriana were harvested at the riverbank and placed in different saline and nutrient solutions in a greenhouse in order to simulate a saline flow. After 7 acclimatization days, plants were exposed to the following saline concentrations: 0, 50, 170 and 250mM NaCl, for a period of 30 days. After this, leaf gas exchange, chlorophyll a fluorescence, Falker chlorophyll index, leaf ultrastructure and micromorphology were evaluated. The highest saline concentration affected mainly the CO2 assimilation, internal CO2 concentration and the intrinsic efficiency of water use. The chlorophyll a fluorescence and Falker index were not significantly affected by any of the saline conditions, however chloroplast was damaged. Concomitantly, leaf salt glands clearly evidenced a higher salt secretion, when plants were submitted to the greatest saline concentration. The results indicated that A. schaueriana tolerates different salt concentration in the mangrove and excretes salt excess by salt gland, and this can attenuate the saline stress. However, when exposed to longer periods of higher salt fluxes the plant growth can be affected.

Molecular, Biochemical and Ultrastructural Changes Induced by Pb Toxicity in Seedlings of Theobroma cacao L.

Pb is a metal which is highly toxic to plants and animals, including humans. High concentrations of Pb have been observed in beans of T. cacao, as well as in its products. In this work, we evaluated the molecular, biochemical, and ultrastructural alterations in mature leaves and primary roots of seedlings of two progenies of T. cacao, obtained from seed germination in different concentrations of Pb (0, 0.05, 0.1, 0.2, 0.4, 0.8 g L(-1)), in the form of Pb(NO3)2. The progenies resulted from self-fertilization of Catongo and a cross of CCN-10 x SCA-6. The Pb, supplied via seminal, caused alterations in the ultrastructures of the mesophyll cells and in the amount of starch grains in the chloroplasts. The dosage of substances reactive to thiobarbituric acid showed that Pb induced lipid peroxidation. The activity of guaiacol peroxidases and the expression of genes associated to synthetase of phytochelatin, SODcyt and PER increased in response to Pb. In addition, there was alteration in the expression of stress-related proteins. The progeny of CCN-10 x SCA-6 was more tolerant to Pb stress when compared to Catongo, since: (i) it accumulated more Pb in the roots, preventing its translocation to the shoot; (ii) it presented higher activity of peroxidases in the roots, which are enzymes involved in the elimination of excess of reactive oxygen species; and (iii) increased expression of the gene in the phytochelatin biosynthesis route. The results of the proteomic analysis were of paramount importance to differentiate the defense mechanisms used by both progenies of T. cacao.

Pb and Cd on growth, leaf ultrastructure and essential oil yield mint (Mentha arvensis L.) / Pb e Cd no crescimento, ultraestrutura foliar e produção de óleo essencial de hortelã (Mentha arvensis L.)

Contamination of medicinal plants with heavy metals as Pb and Cd can affect the growth and the essential oil production of the plants and represent a risk to those who consume as medicine. This study aimed to evaluate the effects of absorption and localization of Pb and Cd on growth, ultrastructural aspects of leaves and essential oil yield and composition of Mentha arvensis, applied on the soil with increasing concentrations (8, 16, 32, 64 and 128mg kg-1). There was a differential absorption of Pb and Cd by M. arvensis mainly concentrated in the roots. Pb was found in small amounts in the leaves while Cd largely exceeded the safety limit without symptoms of toxicity. The ultrastructural analysis revealed the metal accumulation on vesicles surrounding the mitochondria and the presence of electron dense deposits surrounding the mitochondria, nucleus and chloroplasts. Little changes caused by Pb and Cd application were not enough to affect the growth and essential oil yield and composition of M. arvensis .

A contaminação de plantas medicinais com Pb e Cd pode afetar o crescimento e a produção de óleo essencial das plantas e representa um risco para quem as consome como medicamento. Este estudo teve como objetivo avaliar os efeitos da absorção e localização de Pb e Cd sobre o crescimento das plantas, aspectos ultraestruturais de folhas e sobre o rendimento e composição química do óleo essencial de M. arvensis, os quais foram aplicados ao solo em concentrações crescentes (8, 16, 32, 64 and 128mg kg-1). Houve absorção diferencial de Pb e Cd por M. arvensis, que se concentraram principalmente nas raízes. O Pb foi encontrado em baixas concentrações nas folhas, enquanto o Cd excedeu largamente o limite de segurança sem demonstrar sintomas de toxicidade. As análises ultraestruturais revelaram o acúmulo de metais em vesículas ao redor de mitocôndrias e a presença de depósitos eletrodensos ao redor de mitocôndrias, núcleo e cloroplastos. As pequenas mudanças causadas pela aplicação de Pb e Cd não foram suficientes para afetar o crescimento das plantas e o rendimento e composição química do óleo essencial de M. arvensis .

Effects of lead on anatomy, ultrastructure and concentration of nutrients in plants Oxycaryum cubense (Poep. & Kunth) Palla: a species with phytoremediator potential in contaminated watersheds.

Lead (Pb) has been highlighted as a major pollutant of both terrestrial and aquatic ecosystems, causing negative impacts to these environments. The concentration of Pb in plants has increased in recent decades, mainly due to anthropogenic activities. This study has as a hypothesis that the species Oxycaryum cubense (Poep. & Kunth) Palla, abundant in aquatic environments, has the potential to be used a phytoremediator. The plants were grown in a hydroponic system with Pb in increasing concentrations (0, 4, 8, 16 and 32 mg l(-1)) for 15 days. Inductively coupled mass spectrometer (ICP OES) was used to determine the concentration of mineral nutrients and lead. Optical and transmission electron microscopy were used for the analysis of cellular damage induced by lead in roots and leaves. Ultrastructural alterations were observed as disorganization of thylakoids in the chloroplast and disruption of mitochondrial membranes in cells of leaf tissues of plants subjected to increasing Pb concentrations. There was accumulation of Pb, especially in the root system, affecting the absorption and translocation of some mineral nutrients analysed. In roots, there was reduction in the thickness of the epidermis in plants treated with Pb. This species was shown to be tolerant to the Pb concentrations evaluated, compartmentalizing and accumulating Pb mainly in roots. Due to these results, it may be considered a species with phytoremediation capacity for Pb, with potential rizofiltration of this metallic element in contaminated watersheds.