Botanical Preferences of Africanized Bees (Apis mellifera) on the Coast and in the Atlantic Forest of Sergipe, Brazil

Pollen analysis in honey can be used as an alternative method to research into flowers visited by bees in an area. This study aimed to indentify the main floral families in honey from apiaries in the Atlantic Forest and Sergipe state coast. Honey samples from these apiaries were studied, as well as plants that grow around them, which can be used as a source of foraging for bees. The palynological technique was used to compare the pollen content of honey samples with the pollen grains from leaves of plants found in the vicinity of the apiaries to assess whether they had been visited by bees. The results of studies in both sites were similar in terms of incompatibility of families found in the apiary vicinity and honey. Thus, it was possible to observe that in honey samples from the coast and in the remaining Atlantic forest, the number of families was greater than the number of families found in the apiary vicinity, which highlights the diversity of plants visited by bees and a possible expansion of the visited area for food search. This diversity suggests an adaptive foraging behavior to plant resources available in the environment, which may facilitate the pollination of these botanical families and consequently improve their genetic quality.

Compatibility of Beauveria bassiana commercial isolate with botanical insecticides utilized in organic crops in southern Brazil

Products based on botanical insecticides and entomopathogenic fungi have been widely used in organic farming, especially in southern Brazil. Thus, this study investigated, in vitro, the effect of aqueous extracts and commercial formulations of plants with insecticidal activity on Beauveria bassiana. The treatments comprised the botanical insecticides Neempro (azadiractin +3-tigloylazadirachtol), at the concentrations of 0.25, 0.5, 0.75, and 1.0% (v/v), and DalNeem (neem oil emulsifiable), at 0.5, 1.0, 1.5, and 2.0% (v/v) (both commercial formulations of Azadirachta indica (neem)), and the aqueous extracts, at the concentrations of 2.5, 5.0, 7.5, and 10.0% (w/v), of neem seeds, tobacco powder (Nicotiana tabacum), and catigua leaves (Trichilia clausenii). In potato, dextrose, and agar culture medium, the effects of each product on the mycelial growth and the production and viability of conidia of B. bassiana were estimated. According to the adopted compatibility index, the aqueous extracts of neem seeds and leaves catigua, depending on the concentration used, and the botanical insecticide Neempro, were classified as compatible with the entomopathogen, becoming important alternatives to integrate programmes of integrated pest management, especially in organic farming systems.

TOWARDS NEW BOTANICAL PESTICIDES: THE TOXIC EFFECT OF Eremanthus goyazensis (Asteraceae) LEAVES ESSENTIAL OIL AGAINST Brevipalpus phoenicis (Acari: Tenuipalpidae)

This work reports the chemical characterization of Eremanthus goyzensis essential oil and its toxic effect over Brevipalpus phoenicis. The essential oil displayed a major composition of sesquiterpenes (61.87%) including trans-caryophillene (26.81%) and germacrene-D (13.31%). The fumigation test indicated a promising bioactivity over adult B. phoenicis individuals at 24 h (2.03 mu L/L of air) and 48 h (1.08 mu L/L of air) of exposition. A brief discussion of essential oils composition and their singular role on the toxic effect over B. phoenicis is provided here. Our results may contribute to a new and profitable use of a species of Brazilian flora on agribusiness.

Botanical Origin of Protein Sources Used by Honeybees (Apis mellifera) in an Atlantic Forest

Productive and reproductive traits of beehives are influenced by climate and food availability in the region where the bees are reared or maintained, thus honey and pollen storage, egg-laying conditions of the queen as well as comb occupation are subject to seasonal variations. The present study was conducted in the apiary of the Department of Entomology and Acarology, College of Agriculture Luiz de Queiroz, ESALQ/USP, in the municipality of Piracicaba, in an area containing fruit trees, ornamental plants and a fragment of a native forest. The objective was to identify protein sources used by honeybees (Apis mellifera) over a whole year (2010-2011) in remnants of the Atlantic forest, information that can be used in the conservation and restoration of degraded areas. For sample preparation, the acetolysis method was adopted (Eredtman 1952) and the quantitative analysis was performed by counting successive samples of 900 grains per sample which were grouped by botanical species and/or pollen types. The results show that the bees used various plant types in the area, including ruderal species, to maintain their colonies. Apis mellifera seeks food sources in all plants in the surroundings of the apiary, including herbaceous, shrubs, trees, native or introduced. Eucalyptus sp. played an important role as a food source in all seasons due to its wide availability around the apiary and its high flower production. The most frequent pollen types (greater than 10% of the sample) were Anadenanthera sp., Acacia sp, Miconia sp. and Eucalyptus sp. in winter; Philodendron sp., Mikania cordifolia, Parthenium and Eucalyptus sp. in spring; Alternanthera ficoidea, Chamissoa altissima and Eucalyptus sp. in summer; Philodendron sp., Raphanus sp. and Eucalyptus sp. in autumn.

Towards new botanical pesticides: the toxic effect of Eremanthus goyazensis (Asteraceae) leaves essential oil against Brevipalpus phoenicis (Acari: Tenuipalpidae)

This work reports the chemical characterization of Eremanthusgoyzensis essential oil and its toxic effect over Brevipalpus phoenicis. The essential oil displayed a major composition of sesquiterpenes (61.87%) including trans-caryophillene (26.81%) and germacrene-D (13.31%). The fumigation test indicated a promising bioactivity over adult B. phoenicis individuals at 24 h (2.03 µL/L of air) and 48 h (1.08 µL/L of air) of exposition. A brief discussion of essential oils composition and their singular role on the toxic effect over B. phoenicis is provided here. Our results may contribute to a new and profitable use of a species of Brazilian flora on agribusiness.

Self-Organizing Maps of Molecular Descriptors for Sesquiterpene Lactones and Their Application to the Chemotaxonomy of the Asteraceae Family

The Asteraceae, one of the largest families among angiosperms, is chemically characterised by the production of sesquiterpene lactones (SLs). A total of 1,111 SLs, which were extracted from 658 species, 161 genera, 63 subtribes and 15 tribes of Asteraceae, were represented and registered in two dimensions in the SISTEMATX, an in-house software system, and were associated with their botanical sources. The respective 11 block of descriptors: Constitutional, Functional groups, BCUT, Atom-centred, 2D autocorrelations, Topological, Geometrical, RDF, 3D-MoRSE, GETAWAY and WHIM were used as input data to separate the botanical occurrences through self-organising maps. Maps that were generated with each descriptor divided the Asteraceae tribes, with total index values between 66.7% and 83.6%. The analysis of the results shows evident similarities among the Heliantheae, Helenieae and Eupatorieae tribes as well as between the Anthemideae and Inuleae tribes. Those observations are in agreement with systematic classifications that were proposed by Bremer, which use mainly morphological and molecular data, therefore chemical markers partially corroborate with these classifications. The results demonstrate that the atom-centred and RDF descriptors can be used as a tool for taxonomic classification in low hierarchical levels, such as tribes. Descriptors obtained through fragments or by the two-dimensional representation of the SL structures were sufficient to obtain significant results, and better results were not achieved by using descriptors derived from three-dimensional representations of SLs. Such models based on physico-chemical properties can project new design SLs, similar structures from literature or even unreported structures in two-dimensional chemical space. Therefore, the generated SOMs can predict the most probable tribe where a biologically active molecule can be found according Bremer classification.

Phylogeny of Lundia (Bignoniaceae) based on ndhF and PepC sequences

The circumscription of genera belonging to tribe Bignonieae (Bignoniaceae) has traditionally been complex, with only a few genera having stable circumscriptions in the various classification systems proposed for the tribe. The genus Lundia, for instance, is well characterized by a series of morphological synapomorphies and its circumscription has remained quite stable throughout its history. Despite the stable circumscription of Lundia, the circumscription of species within the genus has remained problematic. This study aims to reconstruct the phylogeny of Lundia in order to refine species circumscriptions, gain a better understanding of relationships between taxa, and identify potential morphological synapomorphies for species and major clades. We sampled 26 accessions representing 13 species of Lundia, and 5 outgroups, and reconstructed the phylogeny of the genus using a chloroplast (ndhF) and a nuclear marker (PepC). Data derived from sequences of the individual loci were analyzed using parsimony and Bayesian inference, and the combined molecular dataset was analyzed with Bayesian methods. The monophyly of Lundia nitidula, a species with a particularly complex circumscription, was tested using Shimodaira-Hasegawa (SH) test and the approximately unbiased test for phylogenetic tree selection (AU test). In addition, 40 morphological characters were mapped onto the tree that resulted from the analysis of the combined molecular dataset in order to identify morphological synapomorphies of individual species and major clades. Lundia and most species currently recognized within the genus were strongly supported as monophyletic in all analyses. One species, Lundia nitidula, was not resolved as monophyletic, but the monophyly of this species was not rejected by the AU and SH tests. Lundia sect. Eriolundia is resolved as paraphyletic in all analyses, while Lundia sect. Eulundia is monophyletic and supported by the same morphological characters traditionally used to circumscribe this section. The phylogeny of Lundia contributed important information for a better circumscription of species and served as basis the taxonomic revision of the genus.

Efficiency of neem oil nanoformulations to Bemisia tabaci (GENN.) Biotype B (Hemiptera: Aleyrodidae)

The nanotechnology, through encapsulation of active ingredients, has showed an important way to avoid problems with quickly degradation of the pesticide molecules. Thus, neem (Azadirachta indica) oil nanoformulations containing beta-ciclodextrin and poli-epsilon-caprolactone (PCL) were tested as to their control efficiency against eggs and nymphs of Bemisia tabaci (Genn.) biotype B reared in soybean. The Lethal Concentration (LC50) was estimated using a commercial neem oil (Organic Neem (R)) on first-instar nymphs to establish the adequate volume of the nanoformulations per treatment. After that, they were sprayed on eggs and first-instar nymphs in laboratory and greenhouse and on third-instar nymphs in greenhouse. The commercial neem oil and distilled water were used as controls. Egg viability was not affected by any treatment. Among six nanoformulations, only one was efficient against the first-instar nymphs in laboratory conditions. However, its effective period was not increased as expected. In greenhouse, first-instar nymphs were more affected by two nanoformulations which were significantly different of the commercial neem oil - the most effective one. No mortality differences among the formulations in the third-instar test were observed. The nanoformulations were less efficient to control the B. tabaci biotype B nymphs than the commercial neem oil.

A New Species of Pelexia (Orchidaceae, Spiranthinae) from Sao Paulo, Brazil

A new species of Pelexia Poit. ex Until. (Orchidaceae, Spiranthinae) occurring in central Sao Paulo, southeastern Brazil, is described and illustrated as P. vinosa A. W. C. Ferreira, M. I. S. Lima & Pansarin. Pelexia vinosa is recognized by its leaves that are present at flowering and its dark purple leaf blades with reddish margins. Inflorescences are sparsely pubescent and reddish. The red sepals contrast with the white hyaline petals and labellum. The species is notable for its spurlike nectary that is parallel and adnate to the ovary. The new species is morphologically similar to P. laxa (Poepp. & Endl.) Lindl. In addition, the need to preserve native areas of the interior of Sao Paulo State (habitat of P. vinosa) is discussed.

MICROSATELLITE LOCI FOR TUCUMA OF AMAZONAS (ASTROCARYUM ACULEATUM) AND AMPLIFICATION IN OTHER ARECACEAE

Premise of the study: Microsatellite loci were developed for tucuma of Amazonas (Astrocaryum aculeatum), and cross-species amplification was performed in six other Arecaceae, to investigate genetic diversity and population structure and to provide support for natural populations management. Methods and Results: Fourteen microsatellite loci were isolated from a microsatellite-enriched genomic library and used to characterize two wild populations of tucuma of Amazonas (Manaus and Manicore cities). The investigated loci displayed high polymorphism for both A. aculeatum populations, with a mean observed heterozygosity of 0.498. Amplification rates ranging from 50% to 93% were found for four Astrocaryum species and two additional species of Arecaceae. Conclusions: The information derived from the microsatellite markers developed here provides significant gains in conserved allelic richness and supports the implementation of several molecular breeding strategies for the Amazonian tucuma.

MICROSATELLITE MARKERS DEVELOPED FOR UTRICULARIA RENIFORMIS (LENTIBULARIACEAE)

Premise of the study: Microsatellite markers were developed to study the genetic diversity and population structure of the carnivorous bladderwort Utricularia reniformis, which is endemic to the Atlantic Forest of southern and southeastern Brazil. Cross-species amplification was tested in U. gibba, U. neottioides, U. subulata, and Pinguicula benedicta. Methods and Results: The U. reniformis genome was sequenced in a 454 GS FLX sequencer, and eight primer sets were developed based on the microsatellites identified from the reads. All loci are polymorphic, showing 1.6 to 4.8 alleles per population. Preliminary results show that primer sets are suitable for population-level studies. Cross-species amplifi cation was successful in three other Utricularia species and one Pinguicula species. Conclusions: Markers developed in this study provide tools for analyses of intra- and interpopulation genetic diversity in Utricularia and Pinguicula.

ISOLATION AND CHARACTERISTICS OF EIGHT NOVEL POLYMORPHIC MICROSATELLITE LOCI IN LIPPIA ALBA (VERBENACEAE)

Premise of the study: A set of eight microsatellite (simple sequence repeat [SSR]) markers for Lippia alba, an important medicinal and cosmetic plant, was developed to aid studies of genetic diversity and to define efficient strategies for breeding programs. Methods and Results: Using a (CT)(8)- and (GT)(8)-enriched library, a total of 11 SSR loci were developed and optimized in L. alba. Of the 11 loci, eight were found to be polymorphic after screening 61 accessions from two populations. The parameters used to characterize loci were expected heterozygosity (H-e) and number of alleles. A total of 44 alleles were identified, with an average of 5.5 alleles per loci, which were moderately to highly informative according to H-e. Conclusions: These new SSR markers have potential for informing genetic diversity, allele mining, and mapping studies and will be used to generate information for breeding programs of L. alba

MICROSATELLITE MARKERS FOR PLATHYMENIA RETICULATA (LEGUMINOSAE)

Premise of the study: Microsatellite markers were developed and characterized to investigate genetic diversity and gene flow and to help in conservation efforts for the endangered timber species Plathymenia reticulata. Methods and Results: Eleven microsatellite loci were characterized using 60 adult trees of two populations of P. reticulata from the Atlantic Forest of southern Bahia, Brazil. Of these, nine loci were polymorphic, with an average of 4.39 alleles per locus. The average expected heterozygosity per population ranged from 0.47 to 0.55. The combined exclusion probability was 0.99996. Conclusions: Our results reveal that the microsatellite markers developed in this study are an effective tool for paternity and genetic structure analysis that may be useful for conservation strategies.

Immediate and long-term effects of polysaccharides-based formulations on human skin

A new trend in cosmetic formulations is the use of biotechnological raw materials as the polysaccharides from Klebsiella pneumoniae, which are supposed to enhance cell renewal, improve skin hydration and micro-relief. Botanical extracts of Myrtus communis leaves contain different sugars, which may provide the same benefits. Thus, the objective of this study was to evaluate through objective and subjective analysis the immediate and long-term effects of cosmetic formulations containing polysaccharides biotechnologically-originated and / or the ones contained in Myrtus communis extracts. Three polysaccharide-based and placebo formulations were applied on the forearm skin of 40 volunteers. Skin hydration, transepidermal water loss (TEWL), viscoelasticity and skin micro-relief measurements were made before and 2 hours after a single application and after 15 and 30 day-periods of daily applications. Answers to a questionnaire about perceptions of formulation cosmetic features constituted the subjective analysis. All polysaccharide-based formulations enhanced skin hydration. Formulations with isolated or combined active substances improved skin barrier function as compared to placebo, in the short and long term studies. Formulations containing Myrtus communis extracts had the highest acceptance. Results suggest that daily use of formulations containing these substances is important for protection of the skin barrier function.

GEOGRAPHICAL GENETIC STRUCTURING AND PHENOTYPIC VARIATION IN THE VELLOZIA HIRSUTA (VELLOZIACEAE) OCHLOSPECIES COMPLEX

Premise of the study: Vellozia hirsuta forms a complex presenting wide morphological and anatomical variation, resulting in five specific names and 14 morpho-anatomical patterns occurring in disjunct populations. We carried out a phylogeographical study to investigate the existence of correlation among the genetic and morphological patterns within this complex, and to determine whether it is composed of various species or should be treated as an ochlospecies, a species having widely polymorphic and weakly polytypic complex variation, with morphological characteristics varying independently. Methods: We carried out phylogeographical analyses using cpDNA rpl32F-trnL intergenic region. Key results: We found 20 haplotypes in 23 populations sampled. The populations are genetically structured (Phi(ST) = 0.818) into four phylogeographical groups demonstrating geographical structuring but with no correlation with morpho-anatomical patterns. Our analyses do not support recognizing any of the species now synonymized under Vellozia hirsuta. The northern populations were the most genetically differentiated and could be considered a distinct taxon, as they are also morphologically different. Conclusions: It is recommended that Vellozia hirsuta be considered a single enormously variable species. The patterns of variation within V. hirsuta probably are related to climatic changes that occurred during the Pleistocene Epoch in tropical Brazil when reductions in forest cover favored the expansion of V. hirsuta populations into extensive lowland areas. The expansion of forest cover at the end of the glaciations would have again restricted the occurrence of campos rupestres vegetation to high elevations, which constitute the current centers of diversity of this species.