Antimicrobial activity of Rheedia brasiliensis and 7-epiclusianone against Streptococcus mutans

This in vitro study evaluated the antimicrobial activity of extracts obtained from Rheedia brasiliensis fruit (bacupari) and its bioactive compound against Streptococcus mutans. Hexane, ethyl-acetate and ethanolic extracts obtained (concentrations ranging from 6.25 to 800 mu g/ml) were tested against S. mutans UA159 through MIC/MBC assays. S. mutans 5-days-old biofilms were treated with the active extracts (100 x MIC) for 0, 1, 2, 3 and 4 h (time-kill) and plated for colony counting (CFU/ml). Active extracts were submitted to exploratory chemical analyses so as to isolate and identify the bioactive compound using spectroscopic methods. The bioactive compound (concentrations ranging from 0.625 to 80 mu g/ml) was then tested through MIC/MBC assays. Peel and seed hexane extracts showed antimicrobial activity against planktonic cells at low concentrations and were thus selected for the time kill test. These hexane extracts reduced S. mutans biofilm viability after 4 h, certifying of the bioactive compound presence. The bioactive compound identified was the polyprenylated benzophenone 7-epiclusianone, which showed a good antimicrobial activity at low concentrations (MIC: 1.25-2.5 mu g/ml; MBC: 10-20 mu g/ml). The results indicated that 7-epiclusianone may be used as a new agent to control S. mutans biofilms; however, more studies are needed to further elucidate the mechanisms of action and the anticariogenic potential of such compound found in R. brasiliensis. (C) 2008 Elsevier GmbH. All rights reserved.

Effects of 7-Epiclusianone on Streptococcus mutans and Caries Development in Rats

The aim of this study was to evaluate the effects of 7-epiclusianone (7-epi) on specific virulence attributes of Streptococcus mutans in vitro and on development of dental caries in vivo. 7-Epi was obtained and purified from fruits of Rheedia brasiliensis. We investigated its influence on surface-adsorbed glucosyltransferase (Gtf) B activity, acid production, and viability of S. mutans in biofilms, as well as on caries development using a rodent model. 7-Epi (100 mu g/mL) significantly reduced the activity of surface-adsorbed GtfB (up to 48.0 +/- 1.8 of inhibition at 100 mu g/mL) and glyco-lytic pH-drop by S. mutans in biofilms (125 and 250 mu g/mL) (vs. vehicle control, p < 0.05). In contrast, the test compound did not significantly affect the bacterial viability when compared to vehicle control (15% ethanol, p > 0.05). Wistar rats treated topically with 7-epi (twice daily, 60-s exposure) showed significantly smaller number of and less severe smooth-and sulcal-surface carious lesions (p < 0.05), without reducing the S. mutans viable population from the animals` dental biofilms. In conclusion, the natural compound 7-epiclusianone may be a potentially novel pharmacological agent to prevent and control dental caries disease.

Inhibition of Auxin Transport from the Ovary or from the Apical Shoot Induces Parthenocarpic Fruit-Set in Tomato Mediated by Gibberellins

Fruit-set in tomato (Solanum lycopersicum) depends on gibberellins and auxins (GAs). Here, we show, using the cv MicroTom, that application of N-1-naphthylphthalamic acid (NPA; an inhibitor of auxin transport) to unpollinated ovaries induced parthenocarpic fruit-set, associated with an increase of indole-3-acetic acid (IAA) content, and that this effect was negated by paclobutrazol (an inhibitor of GA biosynthesis). NPA-induced ovaries contained higher content of GA(1) (an active GA) and transcripts of GA biosynthetic genes (SlCPS, SlGA20ox1, and -2). Interestingly, application of NPA to pollinated ovaries prevented their growth, potentially due to supraoptimal IAA accumulation. Plant decapitation and inhibition of auxin transport by NPA from the apical shoot also induced parthenocarpic fruit growth of unpollinated ovaries. Application of IAA to the severed stump negated the plant decapitation effect, indicating that the apical shoot prevents unpollinated ovary growth through IAA transport. Parthenocarpic fruit growth induced by plant decapitation was associated with high levels of GA(1) and was counteracted by paclobutrazol treatment. Plant decapitation also produced changes in transcript levels of genes encoding enzymes of GA biosynthesis (SlCPS and SlGA20ox1) in the ovary, quite similar to those found in NPA-induced fruits. All these results suggest that auxin can have opposing effects on fruit-set, either inducing (when accumulated in the ovary) or repressing (when transported from the apical shoot) that process, and that GAs act as mediators in both cases. The effect of NPA application and decapitation on fruit-set induction was also observed in MicroTom lines bearing introgressed DWARF and SELF-PRUNING wild-type alleles.

Hormonal modulation of photomorphogenesis-controlled anthocyanin accumulation in tomato (Solanum lycopersicum L. cv Micro-Tom) hypocotyls: Physiological and genetic studies

Hormones are likely to be important factors modulating the light-dependent anthocyanin accumulation. Here we analyzed anthocyanin contents in hypocotyls of near isogenic Micro-Tom (MT) tomato lines carrying hormone and phytochrome mutations, as single and double-mutant combinations. In order to recapitulate mutant phenotype, exogenous hormone applications were also performed Anthocyanin accumulation was promoted by exogenous abscisic acid (ABA) and inhibited by gibberellin (GA), in accordance to the reduced anthocyanin contents measured in ABA-deficient (notabills) and GA-constitutive response (procera) mutants. Exogenous cytokinin also enhanced anthocyanin levels in MT hypocotyls. Although auxin-insensitive chageotropica mutant exhibited higher anthocyanin contents, pharmacological approaches employing exogenous auxin and a transport inhibitor did not support a direct role of the hormone in anthocyanin accumulation Analysis of mutants exhibiting increased ethylene production (epwastic) or reduced sensitivity (Never ripe), together with pharmacological data obtained from plants treated with the hormone, indicated a limited role for ethylene in anthocyanin contents. Phytochrome-deficiency (aurea) and hormone double-mutant combinations exhibited phenotypes suggesting additive or synergistic interactions, but not fully espistatic ones, in the control of anthocyanin levels in tomato hypocotyls. Our results indicate that phytochrome-mediated anthocyanin accumulation in tomato hypocotyls is modulated by distinct hormone classes via both shared and independent pathways. (C) 2010 Elsevier Ireland Ltd. All rights reserved

Allelopathic potential of bark and leaves of Esenbeckia leiocarpa Engl. (Rutaceae)

We investigated the inhibitory potential of aqueous extracts of bark and leaves of Esenbeckia leiocarpa Engl. on lettuce germination and early seedling growth. We compared the effects of four concentrations (100, 75, 50 and 25%) of each extract to water and polyethylene glycol (PEG 6000) solution controls for four replicates of 50 seeds tor germination and four replicates of ten seedlings for seedling growth. The inhibitory effects of E. leiocarpa extracts on the percentage of germination and on the germination speed seemed to be inure than simply an osmotic effect, except for the percentage of seeds germinated in bark extracts. When compared to water control. both bark and leaf extracts delayed germination, and leaf extracts also affected the percentage of germinated seeds. Leaf ex tracts of all concentrations strongly inhibited the development of seedlings and caused them some degree of abnormality; bark extracts also caused abnormalities and reduced seedling growth. Root development was more sensitive to the extracts than hypocotyl growth. The negative effects of leaf extracts on germination and seedling growth were more pronounced than those of bark extracts, and the overall effects of both extracts were positively correlated with extract concentrations.

Characterization of electrical penetration graphs of Bucephalogonia xanthophis, a vector of Xylella fastidiosa in citrus

The sharpshooter Bucephalogonia xanthophis (Berg) (Homoptera: Cicadellidae) is a vector of the xylem-limited bacterium, Xylella fastidiosa (Wells, Raju, Hung, Weisburg, Mandelco-Paul, and Brenner), which causes citrus variegated chlorosis. Despite the importance of citrus variegated chlorosis, the probing behavior of vectors on citrus and its implications for transmission of X. fastidiosa have not been studied. Here we studied electrical penetration graph (EPG-DC system) waveforms produced by B. xanthophis on Citrus sinensis (L.) Osbeck (Rutaceae), and their relationships with stylet activities and xylem ingestion. Electrical penetration graph waveforms were described based on amplitude, frequency, voltage level, and electrical origin of the observed traces during stylet penetration on plant tissues. The main waveforms were correlated with histological observations of salivary sheaths in plant tissues and excretion analysis, in order to determine stylet activities and their precise position. Six waveforms and associated activities are described: (S) secretion of salivary sheath and intracellular stylet pathway, (R) resting during stylet pathway, (Xc) contact of stylets with xylem vessels, (Xi) active xylem ingestion, (N) interruption within the xylem phase (during Xc or Xi), and (W) withdrawal of stylet from the plant. The sharpshooter spent 91.8% of its probing time with its stylet in the xylem, where the main activity was ingestion (Xi: 97.5%). During a probe, the most likely sequence of events is secretion of salivary sheath and pathway (S) through epidermal and parenchyma cells (all individuals), followed by contact with xylem (Xc) (67.6% of all individuals) and ingestion (Xi) (88.3% of those that exhibit waveform Xc). The mean time to contact the xylem (Xc) and initiate ingestion (Xi) after onset of the first probe was 27.8 and 34.2 min, respectively. However, sustained xylem ingestion (Xi > 5 min) was established after 39.8 min, on average. This information is basic for future studies on the transmission mechanisms of X. fastidiosa and in order to establish control strategies aimed at interfering with this process.

Ectopic expression of soybean leghemoglobin in chloroplasts impairs gibberellin biosynthesis and induces dwarfism in transgenic potato plants

We have characterized potato (Solanum tuberosum L.) plants expressing a soybean leghemoglobin that is targeted to plastids. Transgenic plants displayed a dwarf phenotype caused by short internode length, and exhibited increased tuberization in vitro. Under in vivo conditions that do not promote tuberization, plants showed smaller parenchymal cells than control plants. Analysis of gibberellin (GA) concentrations indicated that the transgenic plants have a substantial reduction (approximately 10-fold) of bioactive GA(1) concentration in shoots. Application of GA(3) to the shoot apex of the transformed plants completely restored the wild type phenotype suggesting that GA-biosynthesis rather than signal transduction was limiting. Since the first stage of the GA-biosynthetic pathway is located in the plastid, these results suggest that an early step in the pathway may be affected by the presence of the leghemoglobin.

Impact of seedling removal on regenerating community structure of a seasonal semideciduous forest

(Impact of seedling removal on regenerating community structure of a seasonal semideciduous forest). Transplanting seedlings and saplings from natural forests has been considered an alternative to producing saplings of native species for forest restoration purposes, but the possible impact of this procedure on plant community regeneration has not been investigated. This work evaluates the impact of different treatments of shrub and tree-seedling (up to 30 cm) removal from a seasonal semideciduous forest fragment located in southeastern Brazil on the natural regeneration process. Eighty 2x2 m plots were installed in two habitats (forest edge and interior) and submitted to four seedling-removal treatments (I, II - 100% removal with or without soil mixing; III - 50% removal without soil mixing: and IV - control treatment Without seedling removal). Regeneration density and richness were evaluated before treatment as well as 6, 12 and 18 months later. The results were compared among treatments for each evaluation period and among periods within treatments. There were similarities between edge and interior. The natural regeneration process did not improve with soil mixing. Plots submitted to seedling removal partially recovered plant density; however, these plots had lower species richness when compared to the control and to the initial values before treatment. Seedling removal has a negative impact on the regeneration process of low-density species, thus the use of natural regeneration as a sapling source for forest restoration purposes should focus only on high-density species with well-known regeneration strategies and not on the community as a whole.

Novel sources of witches` broom resistance (causal agent Moniliophthora perniciosa) from natural populations of Theobroma cacao from the Brazilian Amazon

Witches` broom is a severe disease of Theobroma cacao L. (cacao), caused by the basidiomycete Moniliophthora perniciosa. The use of resistant cultivars is the ultimate method of control, but there are limited sources of resistance. Further, resistance from the most widely used source (`Scavina 6`) has been overcome after a few years of deployment. New sources of resistance have been intensively searched for in the Amazon basin. Here, we evaluated for witches` broom resistance, cacao accessions from various natural cacao populations originally collected in the Brazilian Amazon. Resistance of 43 families was evaluated under nursery and/or field conditions by artificial or natural infection, respectively, based on disease incidence. Screening for resistance by artificial inoculation under nursery conditions appeared to be efficient in identifying these novel resistance sources, confirmed by natural field evaluation over a nine-year period. The increase in natural field infection of `Scavina 6` was clearly demonstrated. Among the evaluated families with the least witches` broom incidence, there were accessions originally collected from distinct river basins, including the Jamari river (`CAB 0371`; `CAB 0388`; `CAB 0392`; and `CAB 0410`); Acre (`CAB 0169`); Javari (`CAB 0352`); Solimes (`CAB 0270`); and from the Purus river basin, the two most outstanding resistant accessions, `CAB 0208` and `CAB 0214`. The large genetic diversity found in cacao populations occurring at river basins from Acre and Amazonas states, Brazil, increased the chance that the selected resistant accessions would be genetically more dissimilar, and represent distinct sources of resistance to M. perniciosa from `Scavina 6`.

Impacts of land-use pattern on soil water-content variability on the Loess Plateau of China

Long-term vegetation restoration carried out on the slopes of the Loess Plateau of China employed different spatial and temporal land-use patterns but very little is known about the effects of these patterns on soil water-content variability. For this study the small Donggou catchment was selected to investigate soil water-content distributions for three spatial scales, including the entire catchment area, sampling transects, and land-use systems. Gravimetric soil water contents were determined incrementally to a soil depth of 1.20 m, on 10 occasions from April to October, 2007, at approximately 20-day intervals. Results indicated that soil water contents were affected by the six land-use types, resulting in four distinct patterns of vertical distribution of soil moisture (uniform, increasing, decreasing, and fluctuating with soil depth). The soil water content and its variation were also influenced in a complex manner by five land-use patterns distributed along transects following the gradients of five similar slopes. These patterns with contrasting hydrological responses in different components, such as forage land (alfalfa)-cropland-shrubland or shrubland-grassland (bunge needlegrass)-cropland-grassland, showed the highest soil water-content variability. Soil water at the catchment scale exhibited a moderate variability for each measurement date, and the variability of soil water content decreased exponentially with increasing soil water content. The minimum sample size for accurate data for use in a hydrological model for the catchment, for example, required many more samples for drier (69) than for wet (10) conditions. To enhance erosion and runoff control, this study suggested two strategies for land management: (i) to create a mosaic pattern by land-use arrangement that located units with higher infiltration capacities downslope from those with lower soil infiltrabilities; and (ii) raising the soil-infiltration capacity of units within the spatial mosaic pattern where possible.

Native tree seedling growth when fertilized with different amounts of dry sewage sludge and mineral fertilizer

The increasing volume of urban sewage nowadays generates considerable amount of sludge to be disposed of. One environmentally adequate destination could be the application of treated and stabilized sludge (biosolids) to forest plantations as fertilizer and soil conditioner. The purpose of this study was to analyze the feasibility of applying sewage sludge, evaluating its effects on native tree seedlings. The species evaluated were aroeira-pimenteira (Schinus terebinthifolia Raddi), cabreuva-vermelha (Myroxy-Ion peruiferum L. f.), pau-de-viola (Cytarexyllum myrianthum Cham), unha-de-vaca (Bauhinia forficata Link), which are usually planted in forest restoration. Seedlings were cultivated in pots, containing a volume of 4 dm(3) of soil, within a greenhouse. The study was developed in the proximity of Campinas, SP, Brazil, and installed in November, 2003. The design was entirely randomized including seven treatments: control; mineral fertilization; and different doses of sewage sludge (biosolids) complemented with potassium, due to the low concentration of this element in the sludge produced by the wastewater treatment plant of Barueri (Metropolitan region of Sao Paulo city). The results showed that the application of different dosages of biosolids promoted different responses in stem height and biomass production. The treatment with 20 g/dm(3) of dry sewage sludge promoted both the highest growth and the highest seedling biomass production, compared to the control treatment. All native tree species treated with the highest dosage of sewage sludge showed a growth similar to that of mineral fertilization. The seedlings of aroeira-pimenteira, pau-de-viola, and unha-de-vaca, all typical species of the initial succession in natural forest ecosystems, grew and produced more biomass than cabreuva-vermelha, a typical species of the final forest succession.

Why don`t our stands grow even faster? Control of production and carbon cycling in eucalypt plantations

The growth of Eucalyptus stands varies several fold across sites, under the influence of resource availability, stand age and stand structure. We describe a series of related studies that aim to understand the mechanisms that drive this great range in stand growth rates. In a seven-year study in Hawaii of Eucalyptus saligna at a site that was not water limited, we showed that nutrient availability differences led to a two-fold difference in stand wood production. Increasing nutrient supply in mid-rotation raised productivity to the level attained in continuously fertilised plots. Fertility affected the age-related decline in wood and foliage production; production in the intensive fertility treatments declined more slowly than in the minimal fertility treatments. The decline in stem production was driven largely by a decline in canopy photosynthesis. Over time, the fraction of canopy photosynthesis partitioned to below-ground allocation increased, as did foliar respiration, further reducing wood production. The reason for the decline in photosynthesis was uncertain, but it was not caused by nutrient limitation, a decline in leaf area or in photosynthetic capacity, or by hydraulic limitation. Most of the increase in carbon stored from conversion of the sugarcane plantation to Eucalyptus plantation was in the above-ground woody biomass. Soil carbon showed no net change. This study and other studies on carbon allocation showed that resource availability changes the fraction of annual photosynthesis used below-ground and for wood production. High resources (nutrition or water) decrease the partitioning below-ground and increase partitioning to wood production. Annual foliage and wood respiration and foliage production as a fraction of annual photosynthesis was remarkably constant across a wide range of fertility treatments and forest age. In the Brazil Eucalyptus Productivity Project, stand structure was manipulated by planting clonal Eucalyptus all at once or in three groups at three-monthly intervals, producing a stand where trees did not segregate into dominants and one that had strong dominance. The uneven stand structure reduced production 10-15% throughout the rotation.

Secondary metabolites produced by Propionicimonas sp (ENT-18) induce histological abnormalities in the sclerotia of Sclerotinia sclerotiorum

Sclerotinia sclerotiorum is a highly aggressive pathogen that causes great economic losses, especially in temperate climates. Several biological control agents are available, but actinobacteria have seldom been used to control this fungus. Our objective was to evaluate the efficiency and ultrastructural effects of the secondary metabolites produced by the ant-associated actinobacterium Propionicimonas sp. ENT-18 in controlling the sclerotia of S. sclerotiorum. We demonstrated total inhibition of sclerotia treated with 62.5 mu g/10 mu l of an ethyl acetate extract of compounds produced by ENT-18, and calculated an LC(50) of 1.69 mu g/sclerotia. Histological and ultrastructural analysis indicated that the cells of the treated sclerotia were severely damaged, suggesting direct action of the biomolecule(s) produced by the actinobacterium ENT-18 on the cell structure of the medullae and rind cell wall. This is the first report demonstrating a novel property of Propionicimonas sp.-antifungal activity against S. sclerotiorum.

Adenanthera pavonina TRYPSIN INHIBITOR RETARD GROWTH OF Anagasta kuehniella (LEPIDOPTERA: PYRALIDAE)

Anagasta kuehniella is a polyphagous pest that feeds on a wide variety of stored products. The possible roles suggested for seed proteinase inhibitors include the function as a part of the plant defensive system against pest via inhibition of their proteolytic enzymes. In this study, a trypsin inhibitor (ApTI) was purified from Adenanthera pavonina seed and was tested for insect growth regulatory effect. The chronic ingestion of ApTI did result in a significant reduction in larval survival and weight. Larval and pupal developmental time of larvae fed on ApTI diet at 1% was significantly longer; the larval period was extended by 5 days and pupal period was 10 days longer, therefore delaying by up to 20 days and resulting in a prolonged period of development from larva to adult. As a result, the ApTI diet emergence rate was only 28% while the emergence rate of control larvae was 80%. The percentage of surviving adults (%S) decreased to 62%. The fourth instar larvae reared on a diet containing 1% ApTI showed a decrease in tryptic activity of gut and that no novel proteolytic form resistant to ApTI was induced. In addition, the tryptic activity in ApTI -fed larvae was sensitive to ApTI. These results suggest that ApTI have a potential antimetabolic effect when ingested by A. kuehniella. (C) 2010 Wiley Periodicals, Inc.

Fungal pathogens as classical biological control agents against arthropods

Fungal entomopathogens have been used more frequently than other types of pathogens for classical biological control. Among 136 programs using different groups of arthropod pathogens, 49.3% have introduced fungal pathogens (including both the traditional fungi and microsporidia). The most commonly introduced species was Metarhizium anisopliae (Metschnikoff) Sorokin, with 13 introductions, followed by Entomophaga maimaiga Humber, Shimazu & Soper, which was released seven times. The majority of introduction programs have focused on controlling invasive species of insects or mites (70.7%) rather than on native hosts (29.4%). Almost half of the introductions of traditional fungi targeted species of Hemiptera and 75% of the microsporidia introduced have been introduced against lepidopteran species. The United States was the country where most introductions of fungi took place (n = 24). From 1993 to 2007, no arthropod pathogens were released in the US due to the rigorous regulatory structure, but in 2008 two species of microsporidia were introduced against the gypsy moth, Lymantria dispar (L.). Establishment of entomopathogenic fungi in programs introducing traditional fungi was 32.1% and establishment was 50.0% for programs introducing microsporidia. In some programs, releases have resulted in permanent successful establishment with no non-target effects. In summary, classical biological control using fungal entomopathogens can provide a successful and environmentally friendly avenue for controlling arthropod pests, including the increasing numbers of invasive non-native species.