Effect of TiO2 Microparticles in Lettuce (Lactuca sativa L.) Seeds and Seedlings.

The particle size reduction technology is used in several segments, including sunscreens and new techniques and product improvement. One of the main particles used in the sunscreens formulation is titanium dioxide (TiO2). This formulation allows for better characteristics of these products. Perspectives like incorporation of the particles by other biological systems beyond humans and their effects should be observed. This work aimed to evaluate the titanium dioxide microparticles phytotoxicity on Lactuca sativa L. plants through tests of germination, growth, and weight analysis using microscopy techniques: optical microscopy (OM) and scanning electron microscopy (SEM). Some of the results showed cellular and morphological damage, mainly in the roots and 50 mg L-1 TiO2 concentration, confirmed by SEM. Additionally, anatomical damages like vascular bundle disruption and irregularity in the cortex cells were confirmed by SEM. Additionally, anatomical damages were observed on the three main organs (root, hypocotyl, and leaves) evidenced by the OM. Perspectives to confirm new hypotheses of the interaction of nanomaterials with biological systems are necessary.

Anatomical traits related to leaf and branch hydraulic functioning on Amazonian savanna plants.

Amazonian savannas are isolated patches of open habitats found within the extensive matrix of Amazonian tropical forests. There remains limited evidence on how Amazonian plants from savannas differ in the traits related to drought resistance and water loss control. Previous studies have reported several xeromorphic characteristics of Amazonian savanna plants at the leaf and branch levels that are linked to soil, solar radiation, rainfall and seasonality. How anatomical features relate to plant hydraulic functioning in this ecosystem is less known and instrumental if we want to accurately model transitions in trait states between alternative vegetation in Amazonia. In this context, we combined studies of anatomical and hydraulic traits to understand the structure-function relationships of leaf and wood xylem in plants of Amazonian savannas. We measured 22 leaf, wood and hydraulic traits, including embolism resistance (as P50), Hydraulic Safety Margin (HSM) and isotope-based water use efficiency (WUE), for the seven woody species that account for 75% of the biomass of a typical Amazonian savanna on rocky outcrops in the state of Mato Grosso, Brazil. Few anatomical traits are related to hydraulic traits. Our findings showed wide variation exists among the seven species studied here in resistance to embolism, water use efficiency and structural anatomy, suggesting no unique dominant functional plant strategy to occupy an Amazonian savanna. We found wide variation in resistance to embolism (-1.6 ± 0.1 MPa and -5.0 ± 0.5 MPa) with species that are less efficient in water use (e.g. Kielmeyera rubriflora, Macairea radula, Simarouba versicolor, Parkia cachimboensis and Maprounea guianensis) showing higher stomatal conductance potential, supporting xylem functioning with leaf succulence and/or safer wood anatomical structures and that species that are more efficient in water use (e.g. Norantea guianensis and Alchornea discolor) can exhibit riskier hydraulic strategies. Our results provide a deeper understanding of how branch and leaf structural traits combine to allow for different hydraulic strategies among coexisting plants. In Amazonian savannas, this may mean investing in buffering water loss (e.g. succulence) at leaf level or safer structures (e.g. thicker pit membranes) and architectures (e.g. vessel grouping) in their branch xylem.

Acute toxicity of titanium dioxide microparticles in Artemia sp. nauplii instar I and II.

In this study, the toxicity effects of titanium dioxide (MTiO2 ) microparticles on Artemia sp. nauplii instar I and II between 24 and 48 h was evaluated. The MTiO2 were characterized using different microscopy techniques. MTiO2 rutile was used in toxicity tests at concentration of 12.5, 25, 50, and 100 ppm. No toxicity was observed in Artemia sp. nauplii instar I at the time of 24 and 48 h. However, Artemia sp. nauplii instar II toxicity was observed within 48 h of exposure. MTiO2 at concentrations of 25, 50 and 100 ppm was lethal for Artemia sp. with a significant difference (p ≤ .05) in relation to the control artificial sea water with LC50 value at 50 ppm. Analysis of optical and scanning electron microscopy revealed tissue damage and morphological changes in Artemia sp. nauplii instar II. By using confocal laser scanning microscopy, cell damage was observed due to the toxicity of MTiO2 at a concentration of 20, 50, and 100 ppm. The high mortality rate is related to the filtration of MTiO2 by Artemia sp. nauplii instar II due to the complete development of the digestive tract.

Rotenoids from Clitoria fairchildiana R. Howard (Fabaceae) seeds affect the cellular metabolism of larvae of Aedes aegypti L. (Culicidae).

Non-domesticated species may represent a treasure chest of defensive molecules which must be investigated and rescued. Clitoria fairchildiana R. Howard is a non-domesticated Fabacea, native from the Amazonian Forest whose seeds are exquisitely refractory to insect predation. Secondary metabolites from these seeds were fractionated by different organic solvents and the CH2Cl2 fraction (CFD - Clitoria fairchildiana dichloromethane fraction), as the most toxic to 3rd instar Aedes aegypti larvae (LC50 180 PPM), was subjected to silica gel chromatography, eluted with a gradient of CH2Cl2: MeOH and sub fractioned in nine fractions (CFD1 - CFD9). All obtained fractions were tested in their toxicity to the insect larvae. Two rotenoids, a 11α-O-ß-D-glucopyranosylrotenoid and a 6-deoxyclitoriacetal 11-O-n-glucopyranoside, were identified in the mixture of CFD 7.4 and CFD 7.5, and they were toxic (LC50 120 PPM) to 3rd instar Ae. aegypti larvae, leading to exoskeleton changes, cuticular detachment and perforations in larval thorax and abdomen. These C. fairchildiana rotenoids interfered with the acidification process of cell vesicles in larvae midgut and caused inhibition of 55% of V-ATPases activity of larvae treated with 80 PPM of the compounds, when compared to control larvae. The rotenoids also led to a significant increase in the production of reactive oxygen species (ROS) in treated larvae, especially in the hindgut region of larvae intestines, indicating a triggering of an oxidative stress process to these insects.

A new bioinspired peptide on defensin from C. annuum fruits: Antimicrobial activity, mechanisms of action and therapeutical potential.

BACKGROUND: Antimicrobial peptides, natural or synthetic, appear as promising molecules for antimicrobial therapy because of their both broad antimicrobial activity and mechanism of action. Herein, we determine the anti-Candida and antimycobacterial activities, mechanism of action on yeasts, and cytotoxicity on mammalian cells in the presence of the bioinspired peptide CaDef2.1G27-K44. METHODS: CaDef2.1G27-K44 was designed to attain the following criteria: high positive net charge; low molecular weight (<3000 Da); Boman index ≤2.5; and total hydrophobic ratio ≥ 40%. The mechanism of action was studied by growth inhibition, plasma membrane permeabilization, ROS induction, mitochondrial functionality, and metacaspase activity assays. The cytotoxicity on macrophages, monocytes, and erythrocytes were also determined. RESULTS: CaDef2.1G27-K44 showed inhibitory activity against Candida spp. with MIC100 values ranging from 25 to 50 µM and the standard and clinical isolate of Mycobacterium tuberculosis with MIC50 of 33.2 and 55.4 µM, respectively. We demonstrate that CaDef2.1G27-K44 is active against yeasts at different salt concentrations, induced morphological alterations, caused membrane permeabilization, increased ROS, causes loss of mitochondrial functionality, and activation of metacaspases. CaDef2.1G27-K44 has low cytotoxicity against mammalian cells. CONCLUSIONS: The results obtained showed that CaDef2.1G27-K44 has great antimicrobial activity against Candida spp. and M. tuberculosis with low toxicity to host cells. For Candida spp., the treatment with CaDef2.1G27-K44 induces a process of regulated cell death with apoptosis-like features. GENERAL SIGNIFICANCE: We show a new AMP bioinspired with physicochemical characteristics important for selectivity and antimicrobial activity, which is a promising candidate for drug development, mainly to control Candida infections.

Mitochondrial dysfunction associated with ascorbate synthesis in plants.

Mitochondria are the major organelles of energy production; however, active mitochondria can decline their energetic role and show a dysfunctional status. Mitochondrial dysfunction was induced by high non-physiological level of L-galactone-1,4-lactone (L-GalL), the precursor of ascorbate (AsA), in plant mitochondria. The dysfunction induced by L-GalL was associated with the fault in the mitochondrial electron partition and reactive oxygen species (ROS) over-production. Using mitochondria from RNAi-plant lines harbouring silenced L-galactone-1,4-lactone dehydrogenase (L-GalLDH) activity, it was demonstrated that such dysfunction is dependent on this enzyme activity. The capacity of alternative respiration was strongly decreased by L-GalL, probably mediated by redox-inactivation of the alternative oxidase (AOX) enzyme. Although, alternative respiration was shown to be the key factor that helps support AsA synthesis in dysfunctional mitochondria. Experiments with respiratory inhibitors showed that ROS formation and mitochondrial dysfunction were more associated with the decline in the activities of COX (cytochrome oxidase) and particularly AOX than with the lower activities of respiratory complexes I and III. The application of high L-GalL concentrations induced proteomic changes that indicated alterations in proteins related to oxidative stress and energetic status. However, supra-optimal L-GalL concentration was not deleterious for plants. Instead, the L-GalLDH activity could be positive. Indeed, it was found that wild type plants performed better growth than L-GalLDH-RNAi plants in response to high non-physiological L-GalL concentrations.

Influence of chromium and sodium on development, physiology, and anatomy of Conilon coffee seedlings.

Some components found in the composition of the tannery sludge are nutrients for the plants; it can be considered an alternative source of fertilization as they have favorable agronomic characteristics. However, it is reported in some studies that the presence of chromium and sodium in this residue causes physiological and anatomical disturbances that inhibit the development of the plants. The objective of this study was to evaluate the influence of chromium and sodium on the physiology, anatomy, and development of Conilon coffee seedlings grown on substrates produced with tannery sludge and equivalent doses of chromium and sodium. The experiment was carried out in nursery using randomized block design, containing 5 treatments and 7 repetitions. The treatments consisted of the application of a 40% tannery sludge dose and equivalent doses of chromium and sodium mixed with a conventional substrate. Notably, the presence of sodium in the substrate caused greater damage to the plants, negatively influencing the physiology, anatomy, and, consequently, development of the plants, while the presence of chromium suggests that it does not influence much the evaluated characteristics. The treatment with tannery sludge, on the other hand, despite containing the same chromium and sodium contents, revealed a more pronounced negative influence on the physiology, anatomy, and development patterns of the seedlings. This shows that sodium and chromium alone are not the only factors responsible for the lowest growth indicators studied.

Biomass and Leaf Acclimations to Ultraviolet Solar Radiation in Juvenile Plants of Coffea arabica and C. canephora.

Despite the negative impacts of increased ultraviolet radiation intensity on plants, these organisms continue to grow and produce under the increased environmental UV levels. We hypothesized that ambient UV intensity can generate acclimations in plant growth, leaf morphology, and photochemical functioning in modern genotypes of Coffea arabica and C. canephora. Coffee plants were cultivated for ca. six months in a mini greenhouse under either near ambient (UVam) or reduced (UVre) ultraviolet regimes. At the plant scale, C. canephora was substantially more impacted by UVam when compared to C. arabica, investing more carbon in all juvenile plant components than under UVre. When subjected to UVam, both species showed anatomic adjustments at the leaf scale, such as increases in stomatal density in C. canephora, at the abaxial and adaxial cuticles in both species, and abaxial epidermal thickening in C. arabica, although without apparent impact on the thickness of palisade and spongy parenchyma. Surprisingly, C. arabica showed more efficient energy dissipation mechanism under UVam than C. canephora. UVam promoted elevated protective carotenoid content and a greater use of energy through photochemistry in both species, as reflected in the photochemical quenching increases. This was associated with an altered chlorophyll a/b ratio (significantly only in C. arabica) that likely promoted a greater capability to light energy capture. Therefore, UV levels promoted different modifications between the two Coffea sp. regarding plant biomass production and leaf morphology, including a few photochemical differences between species, suggesting that modifications at plant and leaf scale acted as an acclimation response to actual UV intensity.

Linking root and stem hydraulic traits to leaf physiological parameters in Coffea canephora clones with contrasting drought tolerance.

Knowing the key hydraulic traits of different genotypes at early seedling stages can potentially provide crucial information and save time for breeding programs. In the current study we investigated: (1) how root, stem and whole plant conductivities are linked to xylem traits, and (2) how the integrated hydraulic system impacts leaf water potential, gas exchange, chlorophyll a fluorescence and the growth of three coffee cultivars (clones of Coffea canephora Pierre ex Froehner cv. Conilon) with known differences in drought tolerance. The Conilon clones CL 14, CL 5 V and CL 109A, classified as tolerant, moderately tolerant, and sensitive to drought respectively, were grown under non-limiting soil-water supply but high atmospheric demand (i.e., high VPDair). CL 14 and CL 5 V displayed higher root and stem hydraulic conductance and conductivity, and higher whole plant conductivity than CL 109A, and these differences were associated with higher root growth traits. In addition, CL 109A exhibited a non-significant trend towards wider vessels. Collectively, these responses likely contributed to reduce leaf water potential in CL 109A, and in turn, reduced leaf gas exchange, especially during elevated VPDair. Even when grown under well-watered conditions, the elevated VPDair observed during this study resulted in key differences in the hydraulic traits between the cultivars corresponding to differences in plant water status, gas exchange, and photochemical activity. Together these results suggest that coffee hydraulic traits, even when grown under non-water stress conditions, can be considered in breeding programs targeting more productive and efficient genotypes under drought and high atmospheric demand.

Photosynthetic quantum efficiency in south-eastern Amazonian trees may be already affected by climate change.

Tropical forests are experiencing unprecedented high-temperature conditions due to climate change that could limit their photosynthetic functions. We studied the high-temperature sensitivity of photosynthesis in a rainforest site in southern Amazonia, where some of the highest temperatures and most rapid warming in the Tropics have been recorded. The quantum yield (Fv /Fm ) of photosystem II was measured in seven dominant tree species using leaf discs exposed to varying levels of heat stress. T50 was calculated as the temperature at which Fv /Fm was half the maximum value. T5 is defined as the breakpoint temperature, at which Fv /Fm decline was initiated. Leaf thermotolerance in the rapidly warming southern Amazonia was the highest recorded for forest tree species globally. T50 and T5 varied between species, with one mid-storey species, Amaioua guianensis, exhibiting particularly high T50 and T5 values. While the T50 values of the species sampled were several degrees above the maximum air temperatures experienced in southern Amazonia, the T5 values of several species are now exceeded under present-day maximum air temperatures.

Data on leaf structural, physiological and nutritional characteristics of species co-occurring in restinga and semidecidual seasonal forest ecosystems.

This paper presents additional data on the leaf structural, physiological and nutritional characteristics of three species (Maytenus obtusifolia, Manilkara subsericea e Inga laurina), co-occurring in restinga and semideciduous seasonal forest (forest). The data of the leaf structural, physiological and nutritional characteristics were obtained from the three species to identify possible adaptive strategies that could explain the co-occurrence of these species in the restinga and forest. In addition, this data can help identify key functional traits in the plant community of restinga and forests that can be employed in the reestablishment of ecological and edaphic processes in these ecosystems. This work presents data complementary to the published article "Acclimatization capacity of leaf traits of species co-occurring in restinga and seasonal semideciduous forest ecosystems" [1].

Programmed cell death in soybean seed coats.

Seed coat is the tissue which establishes an interface between the seed inner tissues and external environment. Our group has shown that cowpea seed coat undergoes coordinated events of programmed cell death (PCD) during development. In relation to germinating seed coats, little is known on PCD events. The goal here was to investigate the biochemical aspects of germinating soybean seed coat, focusing on proteolytic activities related to PCD. In gel and in solution activity profiles of quiescent and germinating seed coat extracts revealed a complex pattern of caspase- and metacaspase-like cysteine protease activities. Trypsin inhibitor and reserve proteins were revealed as potential substrates for these proteases. A pancaspase inhibitor (z-VAD-CHO) affected the radicle length of seeds germinated under its presence. Ultrastructural analysis showed the absence of cell organelles in all seed coat layers after imbibition, while oligonucleosome fragments peaked at 72 h after imbibition (HAI). Altogether, the data suggest the presence of biochemical PCD hallmarks in germinating soybean seed coat and point to the involvement of the detected protease activities in processes such as reserve protein mobilization and weakening of seed coat.