Phylogenetic systematics of dart-poison frogs and their relatives (Amphibia: Athesphatanura: Dendrobatidae)

The known diversity of dart-poison frog species has grown from 70 in the 1960s to 247 at present, with no sign that the discovery of new species will wane in the foreseeable future. Although this growth in knowledge of the diversity of this group has been accompanied by detailed investigations of many aspects of the biology of dendrobatids, their phylogenetic relationships remain poorly understood. This study was designed to test hypotheses of dendrobatid diversification by combining new and prior genotypic and phenotypic evidence in a total evidence analysis. DNA sequences were sampled for five mitochondrial and six nuclear loci (approximately 6,100 base pairs [bp]; x=3,740 bp per terminal; total dataset composed of approximately 1.55 million bp), and 174 phenotypic characters were scored from adult and larval morphology, alkaloid profiles, and behavior. These data were combined with relevant published DNA sequences. Ingroup sampling targeted several previously unsampled species, including Aromobates nocturnus, which was hypothesized previously to be the sister of all other dendrobatids. Undescribed and problematic species were sampled from multiple localities when possible. The final dataset consisted of 414 terminals: 367 ingroup terminals of 156 species and 47 outgroup terminals of 46 species. Direct optimization parsimony analysis of the equally weighted evidence resulted in 25,872 optimal trees. Forty nodes collapse in the strict consensus, with all conflict restricted to conspecific terminals. Dendrobatids were recovered as monophyletic, and their sister group consisted of Crossodactylus, Hylodes, and Megaelosia, recognized herein as Hylodidae. Among outgroup taxa, Centrolenidae was found to be the sister group of all athesphatanurans except Hylidae, Leptodactyidae was polyphyletic, Thoropa was nested within Cycloramphidae, and Ceratophryinae was paraphyletic with respect to Telmatobiinae. Among dendrobatids, the monophyly and content of Mannophryne and Phyllobates were corroborated. Aromobates nocturnus and Colostethus saltuensis were found to be nested within Nephelobates, and Minyobates was paraphyletic and nested within Dendrobates. Colostethus was shown to be rampantly nonmonophyletic, with most species falling into two unrelated cis- and trans-Andean clades. A morphologically and behaviorally diverse clade of median lingual process-possessing species was discovered. In light of these findings and the growth in knowledge of the diversity of this large clade over the past 40 years, we propose a new, monophyletic taxonomy for dendrobatids, recognizing the inclusive clade as a superfamily (Dendrobatoidea) composed of two families (one of which is new), six subfamilies (three new), and 16 genera (four new). Although poisonous frogs did not form a monophyletic group, the three poisonous lineages are all confined to the revised family Dendrobatidae, in keeping with the traditional application of this name. We also propose changes to achieve a monophyletic higher-level taxonomy for the athesphatanuran outgroup taxa. Analysis of character evolution revealed multiple origins of phytotelm-breeding, parental provisioning of nutritive oocytes for larval consumption (larval oophagy), and endotrophy. Available evidence indicates that transport of tadpoles on the dorsum of parent nurse frogs-a dendrobatid synapomorphy-is carried out primitively by male nurse frogs, with three independent origins of female transport and five independent origins of biparental transport. Reproductive amplexus is optimally explained as having been lost in the most recent common ancestor of Dendrobatoidea, with cephalic amplexus arising independently three times. © American Museum of Natural History 2006.

Carotenoid composition of Brazilian fruits and vegetables

Brazil has a wide diversity of food sources of carotenoids. The updated Brazilian database consists of more than 270 items of fruits, vegetables and their prepared and processed products. The database demonstrates variations due to variety, maturity, production technique, climate and processing. Many of these foods are not found in the US and European databases. Good to rich sources (>20 μg/g) of β-carotene are: acerola, bocaiúva, mango 'Extreme' and tucumã. Sources of both α-carotene and β-carotene are buriti, carrot, Cucurbita moschata 'Menina Brasileira', 'Baianinha' and 'Goianinha', and red palm oil. Commercially produced and uncultivated or semi-cultivated leafy vegetables, C. maxima 'Jerimum Caboclo' and the hybrid Tetsukabuto, cooked broccoli are sources of lutein and β-carotene. The edible Tropaeolum majus flower is especially rich in lutein. Although many fruits have β-cryptoxanthin as principal carotenoid (e.g. caja, nectarine, peach, orange-fleshed papaya, tree tomato), the levels are below 20 μg/g. Good to rich sources of lycopene are guava and guava products, papaya, pitanga and pitanga juice, tomato and tomato products, and watermelon. Sources of zeaxanthin are rare; although the principal carotenoid of piqui, the amount is low, lower than that found in buriti.

Frugivory by phyllostomid bats (Mammalia: Chiroptera) in a restored area in Southeast Brazil

We studied the potential contribution of frugivorous bats to the reestablishment of vegetational diversity in a restored area. We analysed the diets of the bat species and the differences between them in the consumption of fruits of autochtonous and allochthonous species. Planted (autochtonous) species were the basis of diets, especially Solanum mauritianum and Cecropia pachystachya, whereas for allochthonous species we found that Piperaceae to be of particular importance. Carollia perspicillata was the main seed disperser for allochthonous species, and potentially the most important bat in the promotion of vegetation diversity in the study area. Our results suggest that frugivorous bats are especially important in the reestablishment of vegetation in disturbed areas, and that restorarion efforts should focus on the planting of different zoochorous species that would guarantee a high year-round fruit production, thereby facilitating natural plant reestablishment by frugivorous bats in regenerating areas. © 2010 Elsevier Masson SAS.

Potential hydroponics systems for seed potato production in tropical conditions

In tropical conditions such as Brazil, depending on the local altitude, potatoes can be planted and harvested in every month of the year successively, which favors pest and disease incidence, especially aphids transmission viruses. Therefore, obtaining good yield depends on constant pest and disease control and quality potato seed acquisition. One of the main strategies to increase a healthier seed potato multiplication rate and production is the use of hydroponic systems, with or without substrates, in channels, pots or boxes. In 2005 and 2008, researches investigated several hydroponic systems with and without the use of substrates. In the hydroponic systems without substrate, the aeroponic system resulted in the highest multiplication rate, with 47 tubers plant-1 compared to NFT and DFT, 35 and 37 tubers plant-1, respectively. With the use of substrates, the pot system obtained better results, reaching 12 tubers plant-1, followed by the capillary system and boxes, with 8 and 7 tubers plant-1, respectively. Potato seed production was influenced by hydroponics systems. Among hydroponic systems with substrate, the pots were shown as the best option. However, higher yields were obtained in the hydroponics systems without substrate, with the aeroponic system that provided the best results.

Adubação nitrogenada e critérios de amostragem foliar para a cultura da batata

The present study aimed to determine which leave will be displayed and the time of collection of this leave. The treatments consisted on five doses of nitrogen, four times for collection sampling and three types of leaves, arranged in a randomized block design with three repetitions, totalizing one hundred and forty-four experimental units. The leaves collections took place every fifteen days at thirty, forty-five, sixty and seventy five days after budbreak (DAB), collecting the laminated / compound young leaf (first leave), newly mature (second leave) and the mature leave (third leave) from the set of terminal leaflets. For this was installed an experiment with the culture of potato (cv. Atlantic), in Barretos/SP, the period of march the june of 2010. According to this data collected in this study, the best time for collecting the leaves is at 30 days after the budbreak of the potato cultivation containing nitrogen concentrations in the leaves minus heterogeneous of plants from a fraction to another and with values R2 = 0.98 being higher to the all times of harvests. To the diagnostic leaf, the highest determination coefficient was observed in the newly mature (second leaf) with R2 = 0.98. It can be observed that the highest levels of nitrogen were found on the first leaf (39.01 kg-1). Nevertheless, the values were very heterogeneous and did not fit the curve being the second leave (newly mature) the one that best represents the nutritional status of the plant. Therefore the recommendation for nutritional diagnosis will collect the recently matured leaves (2nd leaf) 30 days after budbreak.

Rhizoctonia solani GA-3PT es el principal patógeno asociado con el chancro del tallo y la sarna negra de la papa en Colombia

Stem canker and black scurf diseases of potatoes are caused by the basidiomycetous fungus Tanatephorus cucumeris (ana-morphic species complex Rhizoctonia solani). Tese diseases have worldwide distribution wherever potato is grown but their etiology varies depending on the predominance of distinct R. solani anastomosis groups (AGs) in a particular area. Within the species complex, several AGs have been associated with stem canker or black scurf diseases, including AG-1, AG-2-1, AG-2-2, AG-3, AG-4, AG-5 and AG-9. Tis article reports on the most comprehensive population-based study, providing evidence on the distribution of R. solani AGs in Colombian potato fields. A total of 433 isolates were sampled from the main potato cropping areas in Colombia from 2005 to 2009. Isolates were assigned to AGs by conventional PCR assays using specific primers for AG-3, sequencing of the ITS-rDNA and hyphal interactions. Most of the isolates evaluated were assigned to AG-3PT (88.45%), and a few to AG-2-1 (2.54%). Te remaining isolates were binucleate Rhizoctonia (AG-A, E, and I). Pathogenicity tests on the stems and roots of different plant species, including the potato, showed that AG-3PT affects the stems of solanaceous plants. In other plant species, damage was severe in the roots, but not the stems. AG-2-1 caused stem canker of Solanum tuberosum cv. Capiro and in R. raphanistrum and B. campestris subsp. Rapa plantlets and root rot in other plants. Te results of our study indicated that R. solani AG-3PT was the principal pathogen associated with potato stem canker and black scurf diseases of potatoes in Colombia.

Eficiência de utilização de nutrientes por cultivares de batata

The use of cultivars efficient in nutrient use is essential to increase yield and reduce production costs. The objective of this study was to evaluate absorption and nutrients use efficiency on five potato cultivars. The experiment was conduced in Itaí, São Paulo State, Brazil, in winter cropping season of 2008 on Oxisol. The experimental design was randomized blocks with split-plots and four replications. Plots comprised the five potato cultivars and subplots were established by sampling times, performed at planting and every seven days after emergence. At 97 DAP was calculated the nutrients use efficiency for dry matter production and at 122 DAP were calculated the nutrients use efficiency for fresh tuber and tuber dry matter yield. Mondial and Asterix cultivars showed greater dry matter production, higher fresh tubers yield and nutrients accumulation. Nutrients use efficiency for total dry matter and tubers dry matter, was similar among cultivars, but Ágata was more efficient in nutrients use efficiency for fresh tubers yield.

At least two indigenous species of the Bemisia tabaci complex are present in Brazil

Bemisia tabaci is one of the most important global agricultural insect pests, being a vector of emerging plant viruses such as begomoviruses and criniviruses that cause serious problems in many countries. Although knowledge of the genetic diversity of B. tabaci populations is important for controlling this pest and understanding viral epidemics, limited information is available on this pest in Brazil. A survey was conducted in different locations of São Paulo and Mato Grosso states, and the phylogenetic relationships of B. tabaci individuals from 43 populations sampled from different hosts were analysed based on partial mitochondrial cytochrome oxidase 1 gene (mtCOI) sequences. According to the recently proposed classification of the B. tabaci complex, which employs the 3.5% mtCOI sequence divergence threshold for species demarcation, most of the specimens collected were found to belong to the Middle East-Asia Minor 1 species, which includes the invasive populations of the commonly known B biotype, within the Africa/Middle East/Asia Minor high-level group. Three specimens collected from Solanun gilo and Ipomoea sp. were grouped together and could be classified in the New World species that includes the commonly known A biotype. However, six specimens collected from Euphorbia heterophylla, Xanthium cavanillesii and Glycine maxima could not be classified into any of the 28 previously proposed species, although according to the 11% mtCOI sequence divergence threshold, they belong to the New World high-level group. These specimens were classified into a new recently proposed species named New World 2 that includes populations from Argentina. Middle East-Asia Minor 1, New World and New World 2 were differentiated by RFLP analysis of the mtCOI gene using TaqI enzyme. Taq I analysis in silico also differentiates these from Mediterranean species, thus making this method a convenient tool to determine population dynamics, especially critical for monitoring the presence of this exotic pest in Brazil. © 2012 Blackwell Verlag, GmbH.

Estratégias para produção de mudas de tomateiro utilizando paclobutrazol

The modern tomato production, either for fruit table production or for processing, has required quality seedlings, able to withstand environmental adversities after planting and maintaining the standard genotype productive. The aim of this study was to evaluate the effect of three concentrations of PBZ (0, 50 and 100 mg L-1) and two methods of application (seed treatment and watering seedlings) on seedling emergence and seedling production of tomato growth indeterminate. We adopted a completely randomized in a 2x3 factorial scheme with four replications. We evaluated the seedling emergence, the rate of speed of emergence, seedling height and diameter of hypocotyl and their absolute growth rate (TCA), leaf area, dry weight of shoots, roots and total, and reason leaf area (RAF). Results were subjected to analysis of variance and the mean levels of the factor methods of application compared by Tukey test at 5% and the mean levels of the factor concentrations subjected to regression analysis. The application of paclobutrazol via seedlings watering did not affect seedling emergence, significantly reduced the rate of seedling growth and, consequently, the size of the seedlings without affecting the diameter of hypocotyl, plus allow accumulation of dry roots. A concentration of 50 mg L-1 was shown to be sufficient to obtain seedlings more compact, subject to leaf area, the application via watering, beyond allowing an increase of about 37% in dry weight of roots.

A simple, rapid method for the extraction of whole fire ant venom (Insecta: Formicidae: Solenopsis)

The invasive fire ant Solenopsis invicta is medically important because its venom is highly potent. However, almost nothing is known about fire ant venom proteins because obtaining even milligram-amounts of these proteins has been prohibitively challenging. We present a simple and fast method of obtaining whole venom compounds from large quantities of fire ants. For this, we separate the ants are from the nest soil, immerse them in dual-phase mixture of apolar organic solvent and water, and evaporate each solvent phase in separate. The remaining extract from the aqueous phase is largely made up of ant venom proteins. We confirmed this by using 2D gel electrophoresis while also demonstrating that our new approach yields the same proteins obtained by other authors using less efficient traditional methods. © 2013 Elsevier Ltd.

Effects of liver S9 enzymes on somalargine and solasodine cytotoxicity and mass spectrometric fragmentation

The steroidal glycoalkaloid solamargine and its parent aglycone solasodine, isolated from Solanum palinacanthum, were studied in vitro for cytotoxicity and biotransformation by the hepatic S9 fraction as the metabolic activating system. The MTT uptake assay was used to determine viability after 24 h in RAW 264.7 mouse macrophage-like and SiHa cells exposed to various concentrations of the alkaloids in the presence and absence of the hepatic S9 microsomal fraction. The dose-response curves were established for solamargine and solasodine in the presence and absence of external metabolizing system. From these data, the cytotoxic index (CI50) was calculated with mean values of 7.2 and 13.6 μg/mL for Raw cells and 8.6 and 26.0 μg/mL for SiHa cells, respectively. Mass spectrometry was performed to compare the fragmentation patterns of the alkaloids to predict metabolism by the S9 fraction. The mass spectra demonstrated a distinct fragmentation patterns for solamargine and solasodine after the addition of the S9 fraction. In the present study, we demonstrate that the cytotoxic effect of solamargine and solasodine and their metabolites prepared in vitro by biotransformation with the S9 fraction are comparable. These findings suggest that the metabolic activation system S9 fraction may fail to suppress the cytotoxicity of these alkaloids. © 2013 Springer-Verlag Berlin Heidelberg.

Pulp fruit added to culture medium for in vitro orchid development

As an additive in in vitro culture media, fruits have a great potential for facilitating economical orchid production because of lower technology requirements and the ease of obtaining raw materials to formulate culture media. We studied the in vitro growth of Cattleya bicolor Lindl. grown in a simplified culture medium supplemented with different kinds of fruit pulp. The experimental design was completely randomised, with eight seedlings per replication and ten replications per treatment, for a total of 80 seedlings per treatment. The culture medium was made using 150 g L -1 of pulp (without peel or seed) from the following fruits: ripe Santa Cruz tomatoes (Solanum lycopersicum L.), dwarf bananas (Musa cavendishii L.) of intermediate ripeness, light green chayote (Sechium edule (Jacq.) Sw), ripe papaya (Carica papaya L.) or green coconut (Cocos nucifera L.).The treatment control was MS 50 %. The treatments and the control were kept in a growth chamber for seven months before evaluating seedling survival percentage, shoot height, number of leaves, rooting percentage, root number, root length and dry masses of shoot and roots. The highest percentages of seedling survival were obtained using MS 50 %, banana and coconut medium. The seedling survival and rooting percentages illustrate that it is possible to emphasise the culture medium MS 50% and the culture medium supplemented with coconut on the most traditional culture medium with banana or tomato pulp. For the in vitro development of Cattleya bicolor Lindl., a simplified culture medium supplemented with coconut pulp is the most suitable for use as an alternative to MS 50%. A simplified culture medium supplemented with papaya pulp is not recommended for the in vitro development of Cattleya bicolor Lindl.

Indices of competition and bio-agroeconomic efficiency of lettuce and tomato intercrops in greenhouses

Intercropping is a cropping system for the production of greenhouse vegetables. It uses space more efficiently, thus reducing the cost of production. Intercropping tomato and lettuce has not been studied, but knowledge of the competitive and agroeconomic indices of these vegetables can help in the management of the intercropping system. The objectives of this study were to assess, through biological and agroeconomic indices, the competition between species and the profitability of intercropping tomato and lettuce at different times of transplantation over two growing seasons (autumn-winter and summer-winter) in greenhouse conditions. In autumn-winter, two experiments were conducted with a randomised complete-block design and five replicates. Tomato and lettuce were the main crops in the individual experiments. Treatments were arranged in a factorial of two cropping systems (intercropping and individual crops) with four transplants of the secondary crop (0, 10, 20 and 30 days after) plus an additional treatment (individual main crop). These two experiments were repeated in summer-winter. Tomato was the dominant crop regardless of transplant order. Intercropping systems established with transplants of both species on the same day had higher values of indices of competition and bio-agroeconomic efficiency than systems with longer periods of transplants between main and secondary crops. The intercropping of lettuce and tomato in greenhouses, regardless of transplant time or order, had bio-agroeconomic advantages over individual crops. The transplantation of tomato after lettuce is recommended for greater profitability.

Dragon's blood Croton palanostigma induces genotoxic effects in mice

Ethnopharmacological relevance Dragon's blood is a dark-red sap produced by species from the genus Croton (Euphorbiaceae), which has been used as a famous traditional medicine since ancient times in many countries, with scarce data about its safe use in humans. In this research, we studied genotoxicity and clastogenicity of Croton palanostigma sap using the comet assay and micronucleus test in cells of mice submitted to acute treatment. Material and methods HPLC analysis was performed to identify the main components of the sap. The sap was administered by oral gavage at doses of 300 mg/kg, 1000 mg/kg and 2000 mg/kg. For the analysis, the comet assay was performed on the leukocytes and liver cells collected 24 h after treatment, and the micronucleus test (MN) on bone marrow cells. Cytotoxicity was assessed by scoring 200 consecutive polychromatic (PCE) and normochromatic (NCE) erythrocytes (PCE/NCE ratio). Results and conclusion The alkaloid taspine was the main compound indentified in the crude sap of Croton palanostigma. The results of the genotoxicity assessment show that all sap doses tested produced genotoxic effects in leukocytes and liver cells and also produced clastogenic/aneugenic effects in bone marrow cells of mice at the two higher doses tested. The PCE/NCE ratio indicated no cytotoxicity. The data obtained suggest caution in the use of Croton palanostigma sap by humans considering its risk of carcinogenesis. © 2013 Elsevier B.V. All rights reserved.

Plant-derived essential oils affecting settlement and oviposition of Bemisia tabaci (Genn.) biotype B on tomato

The silverleaf whitefly Bemisia tabaci (Genn.) biotype B (Hemiptera: Aleyrodidae) is an economically important pest of tomatoes Solanum lycopersicum (L.), causing irregular ripening on fruits and transmitting several plant pathogenic geminiviruses. The management of this pest is commonly based on repetitive spraying with synthetic pesticides, causing serious environmental damages and increase of resistance by insect population. In the present study, essential oils from the leaves of Artemisia camphorata Vill., Ageratum conyzoides L., Foeniculum vulgare Mill., Lippia alba (Mill.) N. E. Br., Plectranthus neochilus Schltr., and Tagetes erecta L. were investigated for their possible repellent and oviposition-deterrent effects against B. tabaci biotype B on tomato. In a multi-choice assay, P. neochilus essential oil was the most active repellent and oviposition deterrent. Essential oils of A. conyzoides and T. erecta significantly deterred the female B. tabaci biotype B from laying eggs on treated tomato leaflets compared with the control. (E)-Caryophyllene (30.67 %) and the monoterpenes α-pinene (15.02 %) and α-thujene (11.70 %) were identified as the major constituents of the essential oil of P. neochilus. Our findings demonstrated the potential of essential oil of P. neochilus and other oils in the reduction of settlement and oviposition of B. tabaci biotype B on tomato. © 2012 Springer-Verlag Berlin Heidelberg.