Microwave-assisted solvent extraction and analysis of shikimic acid from plant tissues

A better method for determination of shikimate in plant tissues is needed to monitor exposure of plants to the herbicide glyphosate [N-(phosphonomethyl)glycine] and to screen the plant kingdom for high levels of this valuable phytochemical precursor to the pharmaceutical oseltamivir. A simple, rapid, and efficient method using microwave-assisted extraction (MWAE) with water as the extraction solvent was developed for the determination of shikimic acid in plant tissues. High performance liquid chromatography was used for the separation of shikimic acid, and chromatographic data were acquired using photodiode array detection. This MWAE technique was successful in recovering shikimic acid from a series of fortified plant tissues at more than 90% efficiency with an interference-free chromatogram. This allowed the use of lower amounts of reagents and organic solvents, reducing the use of toxic and/or hazardous chemicals, as compared to currently used methodologies. The method was used to determine the level of endogenous shikimic acid in several species of Brachiaria and sugarcane (Saccharum officinarum) and on B. decumbens and soybean (Glycine max) after treatment with glyphosate. The method was sensitive, rapid and reliable in all cases.

Soil seed bank of plant species as a function of long-term soil management and sampled depth

This study aimed at assessing the level of weed infestation indifferent areas that were submitted to different soil management for 16 years. Four management systems were studied: (1) agriculture only under conventional tillage system; (2) agriculture only under no-till system; (3) crop-livestock integrationcrop-livestock integration; (4) livestock only. These areas were sampled at three soil depths (0-5, 5-10 and 10-15 cm), and soil was stored in plastic pots and taken to a greenhouse, where soil moisture and weight were standardized. Soil was kept near 70% moisture field capacity, being revolved every 20 days when all seedling emerged from soil were counted, identified and collected for dry mass assessment. The soil coverage by weeds, number of weed seedlings and dry mass of the weedy community were assessed. A phytoecological analysis was conducted. Weed composition is differentdifferent among management systems after 16 years. Areas with livestock showed much smaller number of weed species in comparison to systems where only grain crops are grown. The presence of livestock affects the potential of germination of soil seed bank. Agriculture systems are similar in terms of weed composition along soil profile, while systems involving livestock show little relation in what regards such sampled depths. Conservationist models of land exploration contribute to reduce severity of weed species occurrence in the long term.

Adaptive value of ryegrass biotypes with low-level resistance and susceptible to the herbicide fluazifop and competitive ability with the wheat culture

Ryegrass is the main weed in wheat crop, causing yield loss due to competition by environmental resources. The objectives of this study were to estimate the fitness cost of ryegrass biotypes with low-level resistance and susceptible to fluazifop and to investigate the relative competitive ability of these biotypes between themselves and against the crop. Thus, fitness cost and competitive ability experiments were conducted under greenhouse conditions. For the fitness cost experiments, the low-level resistant ryegrass biotypes and those susceptible to fluazifop were used. For competitive ability, the treatments were arranged in a replacement series, with five proportions of the wheat cultivar FUNDACEP Horizonte and the low-level resistant and susceptible ryegrass biotypes 100:0, 75:25, 50:50, 25:75 and 0:100. Competitive analysis was carried out through diagrams applied to the replacement experiments and use of relative competitiveness indices. Variables evaluated were plant height, in the fitness cost experiment, and leaf area and shoot dry biomass in both experiments. The ryegrass biotypes show overall similar fitness cost and competitive ability. The wheat cultivar FUNDACEP Horizonte is superior in competitive ability to the ryegrass biotype with low-level resistance and equivalent to the susceptible biotype.

Sinningia allagophylla(Gesneriaceae): in vitro cultivation of a native plant of the Brazilian cerrado

We used axillary buds as initial explants for hormone interaction studies required for in vitro cultivation of S. allagophylla. Callus production was achieved on gelled Murashige & Skoog medium (MS) supplemented with indole-3-acetic acid (IAA= 0.1 and 0.5 mg.l­1 alone or combined with 6 benzylaminopurine) (BA= 0.01 and 0.1 mg.l-1). A hormone balance between IAA and BA that would encourage shoot bud development was not found. Nodal segments from axenic cultures grown in the presence of cytokinin (0.1 mg.1­1 of BA) without any auxin on MS medium with half-strength macronutrients were used as a standard explant source for subsequent experiments on optimum mineral culture media composition for S. allagophylla in vitro cultivation. We found that explants kept in vitro on gelled Gamborg et al. (B5) mineral composition culture medium showed better shoot and specially root growth than on MS medium. Comparisons of the ammonium and nitrate ratios of MS and B5 media indicate that B5 medium has a substantial reduced ammonium ion when compared to MS medium, as well as a lower total nitrogen level. The growth response pattern obtained in vitro may be evidence of the adaptation of this species to soils of poor mineral composition as found in the Brazilian cerrado, as well as an indication that nitrogen levels play a key role for S. allagophylla growth.

Plant species and habitat structure in a sand dune field in the brazilian Caatinga: a homogeneous habitat harbouring an endemic biota

One dune habitat in the semi-arid Caatinga Biome, rich in endemisms, is described based on plant species composition, woody plant density, mean height and phenology and a multivariate analysis of the micro-habitats generated by variables associated to plants and topography. The local flora is composed mainly by typically sand-dweller species of Caatinga, suggesting the existence of a phytogeographic unity related to the sandy areas in the Caatinga biome, which seems to be corroborated by faunal distribution. Moreover, some species are probably endemic from the dunes, a pattern also found in vertebrates. The plant distribution is patchy, there is no conspicuous herbaceous layer and almost 50% of the ground represents exposed sand. Phenology is not synchronized among species, occurring leaves budding and shedding, flowers development and anthesis, fruits production and dispersion both in rainy and dry seasons. Leaf shedding is low compared to the level usually observed in Caatinga areas and about 50% of the woody individuals were producing leaves in both seasons. Spectrum of dispersal syndromes shows an unexpected higher proportion of zoochorous species among the phanerophytes, accounting for 31.3% of the species, 78.7% of the total frequency and 78.6% of the total density. The habitat of the dunes is very simple and homogeneous in structure and most of environmental variance in the area is explained by one gradient of woody plants density and another of increase of Bromelia antiacantha Bertol. (Bromeliaceae) and Tacinga inamoena (K. Schum.) N.P. Taylor & Stuppy (Cactaceae) toward valleys, which seem to determine two kinds of protected micro-habitats for the small cursorial fauna.

Evolution of circadian organization in vertebrates

Circadian organization means the way in which the entire circadian system above the cellular level is put together physically and the principles and rules that determine the interactions among its component parts which produce overt rhythms of physiology and behavior. Understanding this organization and its evolution is of practical importance as well as of basic interest. The first major problem that we face is the difficulty of making sense of the apparently great diversity that we observe in circadian organization of diverse vertebrates. Some of this diversity falls neatly into place along phylogenetic lines leading to firm generalizations: i) in all vertebrates there is a "circadian axis" consisting of the retinas, the pineal gland and the suprachiasmatic nucleus (SCN), ii) in many non-mammalian vertebrates of all classes (but not in any mammals) the pineal gland is both a photoreceptor and a circadian oscillator, and iii) in all non-mammalian vertebrates (but not in any mammals) there are extraretinal (and extrapineal) circadian photoreceptors. An interesting explanation of some of these facts, especially the differences between mammals and other vertebrates, can be constructed on the assumption that early in their evolution mammals passed through a "nocturnal bottleneck". On the other hand, a good deal of the diversity among the circadian systems of vertebrates does not fall neatly into place along phylogenetic lines. In the present review we will consider how we might better understand such "phylogenetically incoherent" diversity and what sorts of new information may help to further our understanding of the evolution of circadian organization in vertebrates

Centromere domain organization and histone modifications

Centromere function requires the proper coordination of several subfunctions, such as kinetochore assembly, sister chromatid cohesion, binding of kinetochore microtubules, orientation of sister kinetochores to opposite spindle poles, and their movement towards the spindle poles. Centromere structure appears to be organized in different, separable domains in order to accomplish these functions. Despite the conserved nature of centromere functions, the molecular genetic definition of the DNA sequences that form a centromere in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe, in the fruit fly Drosophila melanogaster, and in humans has revealed little conservation at the level of centromere DNA sequences. Also at the protein level few centromere proteins are conserved in all of these four organisms and many are unique to the different organisms. The recent analysis of the centromere structure in the yeast S. pombe by electron microscopy and detailed immunofluorescence microscopy of Drosophila centromeres have brought to light striking similarities at the overall structural level between these centromeres and the human centromere. The structural organization of the centromere is generally multilayered with a heterochromatin domain and a central core/inner plate region, which harbors the outer plate structures of the kinetochore. It is becoming increasingly clear that the key factors for assembly and function of the centromere structure are the specialized histones and modified histones which are present in the centromeric heterochromatin and in the chromatin of the central core. Thus, despite the differences in the DNA sequences and the proteins that define a centromere, there is an overall structural similarity between centromeres in evolutionarily diverse eukaryotes.

Investigation of cytotoxic and mutagenic effects of Malpighia glabra L. (barbados cherry) fruit pulp and vitamin C on plant and animal test systems

Fruits are important sources of nutrients in human diet, and Barbados Cherry (Malpighia glabra L.) is of particular interest due to its high content of antioxidants. Diets rich in fruits and vegetables protect individuals against diseases and cancer, but excessive intake of vitamins may act as pro-oxidant and generate changes in DNA. To evaluate the effect of different in natura (BAN) and frozen (BAF) Barbados Cherry pulp concentrations and synthetic vitamin C in liquid form (VC) on the chromosome level and the cell cycle division, root meristeme cells of Allium cepa L. and bone marrow cells of Wistar rats Rattus norvegicus, were used as test system. In Allium cepa L., BAN, at the highest concentration (0.4 mg.mL-1) and BAF, at the lowest concentration (0.2 mg.mL-1), inhibited cell division, and there was recovery of cell division after the recovery period in water only for BAN. In the Wistar rats, all treatments with Barbados Cherry, either acute or subchronic, were not cytotoxic or mutagenic; only the highest concentration of VC increased significantly the rate of chromosomal abnormalities. The data obtained are important to reinforce the use of Barbados Cherry fruit in the diet.

Tolerance level of green seed in soybean seed lots after storage

The occurrence of green soybean seed due to forced maturation or premature plant death caused by drought or foliar and/or root diseases has been common in several Brazilian production areas. Physiological quality of seed lots with green seed may have their germination and vigor potentials affected and therefore discarded by the grain industry. The objective of this experiment was to determine the maximum tolerated level of green seed in soybean seed lots, which is information of major importance for seed producers when taking the decision whether to sell these lots. Soybean seed of the cultivars CD 206, produced in Ubirata, Parana, and ‘FMT Tucunare,’ produced in Alto Garças, Mato Grosso, were used in the study. Green seed and yellow seed of both cultivars were mixed in the following proportions: 0%, 3%, 6%, 9%, 12%, 15%, 20%, 30%, 40%, 50%, 75% and 100%. Seed quality was evaluated by the germination, accelerated aging, tetrazolium and electrical conductivity tests. The contents of a, b and total chlorophyll in the seed were also determined. A complete randomized block design in a factorial scheme (two cultivars x 12 levels of green seed) was used. Seed quality was negatively affected and chlorophyll contents incremented with the increase in the percentage of green seed. Seed germination, viability and vigor, measured by the accelerated aging test, were not reduced with levels of up to 3% green seed, for both cultivars. Levels above 6% green seed significantly reduced the quality of the seed. The quality of seed lots with 9% or more green seed was significantly reduced to the point that their commercialization is not recommended.

Multi-level governance and social cohesion in the European Union: the assessment of local agents, a study case inside Galicia

Implementing multi-level governance has been a key priority in EU cohesion policy. This study assesses the perceived achievements and shortcomings in implementing European Social Fund by analyzing the deficits and weaknesses as well as the poor participation of local agents who are in direct contact with the beneficiaries in order to design and implement this fund, which is the main financial instrument of EU social policy.

The tale of a Trojan horse or the quest for market access? China and the World Trade Organization

In 2001, China finally joined the WTO. The accession of China was looked forward to by many WTO members and China itself. However, observers had some fears that the Chinese accession would prove to be a Trojan horse, disrupting the working of the WTO. This paper looks into the Chinese accession and its involvement in the WTO Dispute Settlement and argues that these fears seem so far to be unfounded.

Uniformity of liquid distribution in the canopy of the bean plant, using the spectrophotometric analysis

Fungal diseases are important factors limiting common bean yield. White mold is one of the main diseases caused by soil pathogens. The objective of this study was to quantify the distribution of a fungicide solution sprayed into the canopy of bean plants by spectrophotometry, using a boom sprayer with and without air assistance. The experiment was arranged in a 2 x 2 x 2 factorial (two types of nozzles, two application rates, and air assistance on and off) randomized block design with four replications. Air assistance influenced the deposition of solution on the bean plant and yield increased significantly with the increased rate of application and air assistance in the boom sprayer.

Prevalence of periodontal disease in dogs and owners' level of awareness - a prospective clinical trial

Periodontal disease (PD) is widely known among veterinarians for its high prevalence and serious consequences to the dogs. The objective of this study was to assess the occurrence of PD in dogs that live in the micro-region of Viçosa, treated at the Veterinary Hospital of the Federal University of Viçosa (HVT - Hospital Veterinário da Universidade Federal de Viçosa), as well as to assess how aware of this disease dog owners are. In order to do so, all dogs treated at the HVT from March 10th, 2009 to November 30th, 2009, on alternate days, had their oral cavities examined. Medical history data, such as age, type of food, main complaint and owner consent, halitosis, presence of dental calculus, inflammation and gingival recession and tooth loss, were collected. A prevalence of 88.67% was found for PD in dogs referred to the HVT, and 2.67% were referred due to this disease. Of all the owners who participated in the study, 43.83% knew about periodontal disease and of these 17.46% made use of some type of prevention or treatment. Therefore, periodontal disease is highly prevalent and the owners are not aware of the disease. Thus, a dog owner clarification program on periodontal disease is needed in the area where HVT-UFV operates.

Adventitious rooting in cuttings of croton and hibiscus in response to indolbutyric acid and humic acid

Adventitious rooting of ornamental plants can be accelerated by the application of growth regulators, such as auxin. Humic acids, organic matter in soil and organic compounds also have a biostimulant effect. This work evaluated the rooting in cuttings of croton (Codianeum variegatum L. Rumph) and hibiscus (Hibiscus rosa-sinensis L) in response to the application of different concentrations of indolbutyric acid (IBA) and humic acid (HA). The experiment was carried out in a greenhouse. Apical stem cuttings were treated with solutions at concentrations of: 0, 250, 500, 1000, 2000 mg L-1 IBA and 0, 10, 20, 30, 40 mmol L-1 HA carbon isolated from vermicomposting. Forty-five days after the applications, the cuttings were removed from the pots containing carbonized rice hull and the following variables were measured: rooting number, length and width of leaves, fresh and dry matter of root and aerial part and root area. The results were subjected to analysis of variance and the qualitative and quantitative effects of the treatments were compared by contrast and regression, respectively. Regression equations were used to determine the maximum efficiency level of root dry matter according to IBA and HA. Higher accumulation of root dry matter was recorded for the treatments with the doses 579 mg L-1 IBA and 14 mmol L-1 HA and 970 mg L-1 IBA and 50 mmol L-1 HA for root cuttings of croton and hibiscus, respectively. It was found that the application of eiher IBA or HA at the indicated doses accelerates rooting in cuttings of croton and hibiscus and contributes to the formation of vigorous plants.

Production of basic potato seed minitubers in substrate and different nitrogen rates

The optimal dose of nitrogen (N) in potato crop depends on the production system. The objective of this study was to determine the optimal dose of N for the production of basic potato seed minitubers and evaluate the effect of N rates on physiological and nitrogen indices in the youngest fully developed leaf (fourth leaf) and in the oldest leaf of the plants at 60 days after planting. The experiment was conducted in a greenhouse at the Departamento de Fitotecnia da Universidade Federal de Viçosa. The treatments consisted of five N rates (0, 45, 90, 180 and 360 mg dm-3), with 10% of each dose applied at planting and the remainder through irrigation water, daily, for 30 days. The nitrogen rates positively influenced the physiological indices (length, width, leaf area, number of leaves, fresh mass and dry mass) and nitrogen (level and content of N and N-NO³ in the dry mass and SPAD) both in the fourth leaf and in the oldest leaf. Likewise, the N rates positively influenced the number and mass of harvested tubers. The largest number (5.44 tubers/plant) and the maximum mass of tubers (243.5 g/plant) were obtained with 360.0 and 332.9 mg N dm-3, respectively. Therefore, the mass and number of tubers were not optimized by the same N rate. The critical SPAD index was 38.8 in the fourth leaf, which was more sensitive to the effect of N rates than the oldest leaf.