Diaspore dispersal season influences root growth and seed traits, but not germination responses, of sympatric Piper species in a seasonal forest.

Seeds may differ in terms of dormancy, longevity, sensitivity to desiccation and dry mass, according to the timing (dry season/rainy season) of diaspore dispersal. In addition, seasonal variations in temperature and water availability can act as signals of the season during seed development, influencing germination responses and root growth. We evaluated the effects of temperature variations and water availability on germination parameters, root growth and seed traits of four coexisting Piper species in seasonal vegetation that differed in diaspore dispersal timing. Eight temperature treatments (15, 20, 23, 25, 28, 30, 35 °C, and alternate 30 °C-20 °C) and four induced water potentials (0, -0.3, -0.6 and -1.2 MPa) were used. The parameters germination onset, germination percentage (G%), mean germination time (MGT), root elongation, seed longevity during ex situ storage and dry mass of seeds were evaluated. Germination responses observed were independent of the diaspore dispersal timing, such as variations in germination onset, G% and MGT, both in temperature and water availability treatments. In contrast, root elongation, longevity and dry mass of seeds varied according to the time of diaspore dispersal. Our results corroborate the hypothesis that the timing of diaspore dispersal is an important factor in controlling the initial development of seedlings in seasonal vegetation, but not in germination responses. The predominance of negative effects of temperature increases and water deficit on root growth shows that the initial stages of plant development can be strongly impacted by these environmental factors.

Ultrastructural and metabolic disorders induced by short-term cadmium exposure in Avicennia schaueriana plants and its excretion through leaf salt glands.

Environmental cadmium (Cd) sources have increased in mangrove sediments in recent decades, inducing cellular damage to many plants. Avicennia schaueriana is abundant in mangrove sites and has been subject to Cd contamination. The possible effects of Cd toxicity and the structural and physiological disturbances to this plant were studied. Can this plant express early cellular tolerance mechanisms to such metal contamination? Seedlings of A. schaueriana were collected from sites of their natural occurrence, placed in plastic pots containing nutrient solution for 60 days, and subsequently exposed to increasing Cd concentrations for 5 days under experimental conditions. The anatomical, ultrastructural and physiological changes induced by Cd were analysed. Cd accumulated mainly in the root system and in pneumatophores, stems and leaves, induced differential accumulation of mineral nutrients, but did not induce necrosis or changes in leaf anatomy. However, there was a decrease in starch grains and an increase in deposited electron-dense material in the cortex and vascular bundles. Cd induced both increases in calcium (Ca) content in shoots and Ca oxalate crystal precipitation in leaf mesophyll and was detected in crystals and in the secretion of salt glands. Our observations and experimental results provide evidence of Cd tolerance in A. schaueriana. As a new feature, despite the clear cellular physiological disorders, this plant is able to eliminate Cd through leaf salt glands and immobilise it in Ca crystals, representing fast mechanisms for Cd exclusion and complexation in leaves in heavy metal coastal polluted marine ecosystems.

The increase of current atmospheric CO2 and temperature can benefit leaf gas exchanges, carbohydrate content and growth in C4 grass invaders of the Cerrado biome.

Leaf gas exchanges, carbohydrate metabolism and growth of three Brazilian Cerrado invasive African grasses were evaluated after growing for 75 days under doubled CO2 concentration and temperature elevated by 3 °C. Results showed that although the species presented photosynthetic C4 metabolism, they all had some kind of positive response to increased CO2. Urochloa brizantha and Megathyrsus maximus showed increased height for all induced environmental conditions. Urochloa decumbens showed only improvement in water use efficiency (WUE), while U. brizantha showed increased CO2 assimilation and M. maximus presented higher biomass accumulation under doubled CO2 concentration. The most significant improvement of increased CO2 in all three species appears to be the increase in WUE. This improvement probably explains the positive increase of photosynthesis and biomass accumulation presented by U. brizantha and M. maximus, respectively. The increase in temperature affected leaf carbohydrate content of M. maximus by reducing sucrose, glucose and fructose content. These reductions were not related to thermal stress since photosynthesis and growth were not harmed. Cellulose content was not affected in any of the three species, just the lignin content in U. decumbens and M. maximus. All treatments promoted lignin content reduction in U. brizantha, suggesting a delay in leaf maturation of this species. Together, the results indicate that climate change may differentially promote changes in leaf gas exchanges, carbohydrate content and growth in C4 plant species studied and all of them could benefit in some way from these changes, constituting a threat to the native Cerrado biodiversity.

Pectin and cellulose cell wall composition enables different strategies to leaf water uptake in plants from tropical fog mountain.

Leaf water uptake (LWU) has been observed in plants of different ecosystems and this process is distinct among different species. Four plant species from the Brazilian fog mountain fields were evaluated in order to detect if leaf water uptake capacity is related to the cell wall composition of leaf epidermis. LWU measurements and their relation to anatomical and biochemical traits were analyzed. Cell wall composition was verified through immunocytochemistry using monoclonal antibodies recognizing pectin compounds, and histochemistry with calcofluor white to track cellulose. Differences in LWU among the four species were clearly revealed. Two species presented higher maximum leaf water content and the lowest values of water absorption speed. The other two species presented opposite behavior, namely, low leaf water uptake and the highest values of water absorption speed. The anatomical traits associated with the cell wall composition corroborated the data on the different LWU strategies. The species with abundant detection of cellulose in their epidermal cell walls absorbed more water, but more slowly, while those with abundant detection of pectins absorbed water at a higher speed. These results indicate that cell wall composition regarding pectin and cellulose are significant for water uptake by the leaf epidermis. Pectin provides greater porosity and absorption speed, while cellulose provides greater hydrophilicity and greater water uptake capacity. Current data indicate that the composition of epidermal cell walls is a relevant trait for leaf water uptake.

Estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (Pohl) Baill. (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro / Toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extract of Jatropha mollissima (Pohl) Baill (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeastern Brazil

RESUMO A cada dia, cepas bacterianas estão tornando-se resistentes a diversos antibióticos, o que faz necessária a busca de novas substâncias eficazes para o tratamento de doenças. Desta forma, este trabalho reporta o estudo preliminar toxicológico, antibacteriano e fitoquímico do extrato etanólico das folhas de Jatropha mollissima (pinhão-bravo, Euphorbiaceae), coletada no Município de Tauá, Ceará, Nordeste Brasileiro. Inicialmente, realizou-se o teste de toxicidade do extrato contra Artemia salina. Na sequencia, foi realizado o ensaio antibacteriano contra quatro cepas bacterianas Gram-negativas (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883) e uma cepa Gram-positiva (Enterococcus faecalis ATCC 29212). Finalmente, fez-se a análise fitoquímica preliminar do extrato ativo para detecção das principais classes de metabólitos especiais. Como resultado, o extrato etanólico das folhas de J. mollissima se mostrou tóxico para Artemia salina, pois apresentou CL50 igual a 406,02 μg/mL. Quanto à ação antibacteriana, o extrato se mostrou ativo contra a bactéria Gram-positiva Enterococcus faecalis ATCC 29212, apresentando moderada atividade antibacteriana (halo de inibição igual a 7,03 mm). Evidenciou-se no extrato bioativo a presença de cumarinas, fenóis, taninos, flavonoides (flavonóis e flavanonas), alcaloides e esteroides, ambas as classes reportadas como antimicrobianos. Portanto, esse extrato tem potencial para ser usado na produção de fármacos contra infecções causadas por bactérias Gram-positivas. No entanto, as informações direcionam estudos futuros para o isolamento e identificação dos compostos bioativos, monitorados sob a ação antibacteriana mais expressiva.

ABSTRACT Each day, bacterial strains are becoming more resistant to various antibiotics, which requires the search for new effective substances for the treatment of diseases. Thus, this study reports the toxicological, antibacterial, and phytochemical preliminary study of the ethanolic extracts of Jatropha mollissima (pinhão-bravo, Euphorbiaceae) leaves, collected in Tauá, Ceará, Northeast of Brazil. Initially, we performed the toxicity testing of the extract against Artemia salina. Then, we conducted the antibacterial assay against four Gram-negative bacterial strains (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Hafnia alvei ATCC 51873, Klebsiella pneumoniae ATCC 13883), and one Gram-positive strain (Enterococcus faecalis ATCC 29212). Finally, we carried out the preliminary phytochemical analysis of the active extract to detect the main classes of special metabolites. As a result, the ethanolic extract of J. mollissima leaves was toxic to Artemia salina, because it presented LC50 equal to 406.02 µg/mL. Regarding antibacterial action, the extract was active against the Gram-positive bacteria Enterococcus faecalis ATCC 29212, with moderate antibacterial activity (inhibition zone equal to 7.03 mm). The bioactive extract had the presence of coumarins, phenols, tannins, flavonoids (flavanols and flavonones), alkaloids and steroids, both classes reported as antimicrobials. Therefore, this extract has the potential to be used in the production of drugs against infections caused by Gram-positive bacteria. However, these information require further studies for the isolation and identification of bioactive compounds, monitored under the more expressive antibacterial action.

Aluminium localization and toxicity symptoms related to root growth inhibition in rice (Oryza sativa L.) seedlings.

We correlated root growth inhibition with aluminium (Al(3+)) localization and toxicity symptoms in rice roots using seedlings of two genotypes (tolerant and sensitive) that were exposed to different AlCl(3) concentrations. Al(3+) localization was evaluated by hematoxylin in primary roots and by morin in cross-sections of the root tips. Neutral invertase enzyme activity and callose (1 -- 3, beta-D-glucan) accumulation were observed and compared with Al(3+) accumulation sites. Root growth was inhibited by Al(3+) in a concentration-specific manner and proportional to the increase of hematoxylin staining, being more pronounced in the sensitive genotype. Morin staining showed the presence of Al(3+) deep within the roots of the sensitive genotype, indicating that the metal was able to penetrate beyond the first few cell layers. In the tolerant genotype, Al(3+) penetration was restricted to the first two cell layers. Ruptures in exodermis and epidermis layers by lateral root protrusions in both genotypes allowed Al3+ to enter into the roots. More intense activity of invertase in roots of the tolerant genotype was also observed, which could be related to greater root growth of this cultivar when submitted to Al(3+) stress. Moreover, Al(3+)-induced callose accumulation was a late response occurring in the same areas where Al(3+) was present.

Ethyl acetate extract and its major constituent, isorhamnetin 3-O-rutinoside, from Nitraria retusa leaves, promote apoptosis of human myelogenous erythroleukaemia cells.

OBJECTIVE: Fractionation of ethyl acetate extract (EA) obtained from Nitraria retusa leaves was assessed using different methods of chromatography, and isorhamnetin3-O-rutinoside (I3-O-R) was isolated from this extract. Its structure was determined using data obtained from (1) H and (13) C NMR spectra, as well as by various correlation experiments (COSY, HMQC and HMBC). Both EA extract and I3-O-R were investigated for their ability to induce apoptosis in human chronic myelogenous erythroleukaemia cells (K562). MATERIALS AND METHODS: Apoptosis of cells from the K562 line was detected by DNA fragmentation, PARP cleavage and by evaluating activities of caspases 3 and 8. RESULTS: Apoptosis, revealed by DNA fragmentation and PARP cleavage, was observed after 48-h incubation of these human myelogenous erythroleukaemia cells (K562), with the tested products. Likewise, caspase 3 and caspase 8 activities were induced in the presence of the EA extract and I3-O-R after 48 h of incubation. CONCLUSION: Our results strongly suggest the involvement of the extrinsic pathway of apoptosis in cells treated by both the original EA extract and its major component, I3-O-R.

[Efflux pumps: their role in Staphylococcus aureus antibiotic resistance]. / Les phénomènes de résistance aux antibiotiques liés aux pompes à efflux : exemple de Staphylococcus aureus.

Efflux pumps located in the bacterial membranes are responsible for low level resistance to antibiotics, considered not to be relevant in the clinic and thus often neglected. However, these pumps contribute to the emergence of high level antibiotic resistance mechanisms, which are responsible for severe complications during the treatment of infectious diseases. Therefore it is necessary to take into account these pumps while developing novel antibacterial agents. Among these new research strategies, the development of efflux pump inhibitors seems to be an attractive approach to restore the activity of some "classical" antibiotics and to limit the emergence of multiresistant strains associated with hospital-acquired infections. In this review, we focalise on Staphylococcus aureus efflux pumps and their potential inhibitors.

New dihydrophenanthrene and phenyldihydroisocoumarin constituents of Trema orientalis.

In the course of the chemical investigation of extracts of the trunk bark and root bark of Trema orientalis, three new compounds were isolated, namely, (9S*,10S*)-3-[7-(3,10-dihydroxy-9-hydroxymethyl-2,5-dimethoxy)-9,10-dihydrophenanthrenyl]propenal (1), (9S*,10S*)-3-[7-(5-O-beta-glucopyranosyl-10-hydroxy-9-hydroxymethyl-2,6-dimethoxy)-9,10-dihydrophenanthrenyl]propenal (2), and (3R*,3aR*,4R*,5S*)-6-O-alpha-arabinopyranosyl-8-hydroxy-3-(4-hydroxyphenyl)-4-(4-hydroxyphenyl)-5-(3,5-dihydroxyphenyl)-3,3a-dihydrocyclopenta[1,2,3-de]isobenzopyran-1-one (3, orientoside A). The structures of 1-3 were determined by spectral methods.

Prenylated xanthones as potential P-glycoprotein modulators.

Dimethylallyl (DMA) derivatives of a naturally occurring xanthone (decussatin 1) were prepared. Their activity as potential P-glycoprotein inhibitors was monitored by affinity of direct binding and compared to that of corresponding DMA-flavones. Both classes of compounds exhibited the same structure-activity relationships. Decreasing polarity enhanced the binding affinity for the P-glycoprotein C-terminal cytosolic domain since DMA derivatives were more active, but unsubstituted hydroxyl group close to the carbonyl was required for efficient activity.

Use of SIMS microscopy and electron probe X-ray microanalysis to study the subcellular localization of aluminium in Vicia faba roots cells.

Received January 4, 1999; Accepted March 25, 1999 Secondary ion mass spectrometry (SIMS), electron probe X-ray microanalysis (EPMA) and transmission electron microscopy (TEM) were used to study the tissular distribution and subcellular localization of aluminium (Al) precipitate in roots of Viciafaba. The broad bean plant, grown in nitrate solution with 193 microM Al3+ at pH 4.8, for 15 days showed Al deposits in the roots. Al accumulation was not detected in the stems nor in the leaves. Al was found mainly localized on the root's surfaces and within the cell walls of the cortical cells. Al signal was not detected in the vascular tissues. Two weeks exposure to Al caused ultrastructural changes in cortical cells and sometimes a complete disruption of these cells. Deposition of Al in form of insoluble complexes associated with phosphorus, appeared as electron opaque materials in the vacuoles of disrupted cortex cells and in the intercellular inclusions. The leaves turned yellowish at the end of 15 days exposure. The use of electron microprobe, to investigate the same tissues as the ones investigated by SIMS, provided complementary results on aluminium allocation.