Three New Polyprenylated Benzophenone Derivatives Isolated from Clusia burle-marxii.

Three new polyprenylated benzophenone derivatives named burlemarxiones G-I (1-3) were isolated from C. burle-marxii trunks (compound 1) and leaves (compounds 2 and 3), along with the known compound burlemarxione F. Burlemarxione G (1) was isolated after methylation with diazomethane and it is the keto-enol tautomeric pair of burlemarxione F. Burlemarxione H (2) derives from burlemarxiones F and G, but it has additional rings due to cyclization of the prenyl group attached to C-5 that establishes new single bonds between C-1 and C-23, as well as, between C-24 and C-29. Burlemarxione I (3) has two additional cyclizations: the first encompasses the cyclization of the former isopentenyl group into an 11,11-dimethyl-six-membered ring, whereas the second produces additional rings due to the cyclization of the prenyl group attached to C-5 that establishes new single bonds between C-1 and C-23, as well as, between C-24 and C-29. All three compounds showed moderate anti-glioma activity. These results show that C. burle-marxii is an important source of sophisticated polyprenylated benzophenone derivatives.

Flavonoids, biphenyls and xanthones from the genus Clusia: chemistry, biological activities and chemophenetics relevance.

Clusia is one of the most important genera of the Clusiaceae family, comprising up to 400 species. This review describes the identification of twenty-two flavonoids from Clusia species, which includes five flavonols (1-4 and 11), six flavones (5-10), one catechin (12), one flavanone (13), and nine biflavonoids (14-22). O- and C-glycosylation are frequently observed amongst these flavonoids. Furthermore, seven biphenyls (23-29) and nine xanthones (30-38) have been isolated from Clusia species. Biphenyls and xanthones show limited occurrence within the genus, but together with biosynthetic insights, they might offer important chemophenetics leads for the consolidation of the genus Clusia within the Clusiaceae family. Altogether, this work provides an overview of the chemistry of the genus Clusia in terms of flavonoids, biphenyls and xanthones, as well as it discusses biological activities and chemophenetics of the isolated compounds, when appropriate.

A multiplex approach of MS, 1D-, and 2D-NMR metabolomics in plant ontogeny: A case study on Clusia minor L. organs (leaf, flower, fruit, and seed).

INTRODUCTION: The genus Clusia L. is mostly recognised for the production of prenylated benzophenones and tocotrienol derivatives. OBJECTIVES: The objective of this study was to map metabolome variation within Clusia minor organs at different developmental stages. MATERIAL AND METHODS: In total 15 organs/stages (leaf, flower, fruit, and seed) were analysed by UPLC-MS and 1H- and heteronuclear multiple-bond correlation (HMBC)-NMR-based metabolomics. RESULTS: This work led to the assignment of 46 metabolites, belonging to organic acids(1), sugars(2) phenolic acids(1), flavonoids(3) prenylated xanthones(1) benzophenones(4) and tocotrienols(2). Multivariate data analyses explained the variability and classification of samples, highlighting chemical markers that discriminate each organ/stage. Leaves were found to be rich in 5-hydroxy-8-methyltocotrienol (8.5 µg/mg f.w.), while flowers were abundant in the polyprenylated benzophenone nemorosone with maximum level detected in the fully mature flower bud (43 µg/mg f.w.). Nemorosone and 5-hydroxy tocotrienoloic acid were isolated from FL6 for full structural characterisation. This is the first report of the NMR assignments of 5-hydroxy tocotrienoloic acid, and its maximum level was detected in the mature fruit at 50 µg/mg f.w. Seeds as typical storage organ were rich in sugars and omega-6 fatty acids. CONCLUSION: To the best of our knowledge, this is the first report on a comparative 1D-/2D-NMR approach to assess compositional differences in ontogeny studies compared with LC-MS exemplified by Clusia organs. Results derived from this study provide better understanding of the stages at which maximal production of natural compounds occur and elucidate in which developmental stages the enzymes responsible for the production of such metabolites are preferentially expressed.

Effects of "Bacuri" Seed Butter (Platonia insignis Mart.), a Brazilian Amazon Fruit, on Oxidative Stress and Diabetes Mellitus-Related Parameters in STZ-Diabetic Rats.

This study aimed to investigate the effects of oral administration of Platonia insignis Mart. ("bacuri") seed butter (BSB) on oxidative stress and diabetes mellitus-related parameters in streptozotocin-induced (STZ) diabetic rats. Diabetes mellitus was induced in female Wistar rats (180-250 g) by the intraperitoneal administration of STZ (45 mg/kg, b.w). BSB (25, 50, and 100 mg/kg) was administered to animals for four weeks. The effect on weight gain, food intake, blood glucose, glycated hemoglobin, hepatic transaminases, plasma and liver TBARS and MPO activity, erythrocyte SOD activity, non-protein sulfhydryl groups (SH-NP), and histopathology of the liver tissue was investigated. BSB at the dose of 100 mg/kg had a positive effect on the reduction in glycated hemoglobin percentage and increased albumin concentration, as well as decreased ALT and AST levels and increased SH-NP liver levels in treated animals compared to normal control rats. Moreover, BSB had no effects on weight gain, food intake, and fasting glucose. Thus, the BSB presented marked properties in improvement of hepatic antioxidant defenses, which demonstrates BSB as a potential hepatoprotective agent in metabolic disorders.

Novel polyprenylated benzophenone derivatives from Clusia burle-marxii.

Three new caged polyprenylated benzophenone derivatives named burlemarxiones D-F (1-3) were isolated from the hexane extract of Clusia burle-marxii trunks. Burlemarxione D (1) contains the tetracyclo[8.3.1.03,11.05,10]tetradecane core skeleton also observed for burlemarxione A, its probable immediate precursor. However, two additional rings are formed to produce an unprecedented complex-caged core skeleton. These additional rings could be formed by a radical cyclization reaction of one prenyl group at C-5 with C-1 and C-33, followed by oxidative dehydrogenation (rearomatization) or by an intramolecular [4 + 2] radical cycloaddition (Diels-Alder reaction), followed by an enolization reaction (rearomatization). Burlemarxiones E and F were isolated after methylation with diazomethane that was necessary to avoid the interconversion of the pair of ß-diketones in tautomeric equilibrium. The proposed biosynthetic pathway for burlemarxiones D-F involves the condensation of either lavandulyl pyrophosphate or 2-(1-methylvinyl)-hexa-5-enyl pyrophosphate with the acylphloroglucinol derivative 6-benzoyl-5-hydroxy-5-cyclohexen-1,3-dione, followed by consecutive prenylation reactions. Therefore, Clusia burle-marxii reinforces the claim that the genus Clusia is an important source of sophisticated caged polyprenylated benzophenone derivatives.

Calanolides E1 and E2, two related coumarins from Calophyllum brasiliense Cambess. (Clusiaceae), displayed in vitro activity against amastigote forms of Trypanosoma cruzi and Leishmania infantum.

In the present work, the MeOH extract from stem barks of Calophyllum brasiliense Cambess. (Clusiaceae) displayed activity against amastigote forms of Trypanosoma cruzi and Leishmania infantum and was subjected to a bioactivity-guided fractionation to give two related coumarins - calanolides E1 (1) and E2 (2). Compounds 1 and 2 were actives to T. cruzi with EC50 values of 12.1 and 8.2 µM, respectively. When tested against L. infantum, the EC50 values were 37.1 and 29.1 µM, respectively. Compound 2, corresponding to anti isomer, showed the best selectivity index (SI) with values >24.4 to T. cruzi and >6.9 to L. infantum in comparison to the syn isomer 1. Furthermore, using an in silico multi-parametric prediction, both compounds did not contain any PAINS sub-structures. Therefore, these data suggest that coumarins 1 and 2 may contribute as scaffolds for the design of novel drug candidates for leishmaniasis and Chagas disease.

Atividades farmacológicas da manteiga de bacuri (Platonia insignis Mart): revisão integrativa / Pharmacological actions of bacuri butter (Platonia insignis Mart): an integrative review

RESUMO Objetivo identificar as atividades farmacológicas da manteiga de bacuri (Platonia insignis Mart.). Métodos revisão integrativa, realizada nas bases de dados Literatura Latino-americana e do Caribe em Ciências da Saúde, Cumulative Index to Nursing and Allied Health Literature, EMBASE, MEDLINE/PubMed, Web of Science, Cochrane Library e SCOPUS, sem delimitação temporal e de idioma. A seleção se constituiu de 13 ensaios pré-clínicos. A avaliação das informações ocorreu de forma descritiva, confrontando com os achados pertinentes. Resultados observou-se que 50,0% das publicações foram indexadas na MEDLINE/PubMed, maioria das publicações ocorreram na Inglaterra (61,5%), seguidas do Brasil e dos Estados Unidos, ambos com 13,3%. Destaca-se que 100,0% dos artigos foram ensaios pré-clínicos; atividades farmacológicas para antioxidante (38,4%) e antileishmanicidas (30,7%). Registrou-se que 38,4% dos ensaios apresentaram testes de toxicidade. Conclusão a manteiga de bacuri (Platonia insignis Mart.) apresentou atividades farmacológicas em ensaios pré-clínicos, como antioxidantes, antileshimaniose, anticonvulsivante e cicatrização de feridas.

ABSTRACT Objective to identify the pharmacological activities of bacuri butter (Platonia insignis Mart.). Methods an integrative review, carried out in the databases of Latin American and Caribbean Literature in Health Sciences, Cumulative Index to Nursing and Allied Health Literature, EMBASE, MEDLINE/PubMed, Web of Science, Cochrane Library and SCOPUS, without the time and language restriction. The selection consisted of 13 pre-clinical trials. The information assessment descriptively took place, comparing with the pertinent findings. Results it was observed that 50.0% of the publications were indexed in MEDLINE/PubMed, most publications were from England (61.5%), followed by Brazil and the United States, both with 13.3%. It is noteworthy that 100.0% of the articles were pre-clinical trials; pharmacological activities for antioxidants (38.4%) and antileishmanicides (30.7%). It was found that 38.4% of the trials presented toxicity tests. Conclusion bacuri butter (Platonia insignis Mart.) Showed pharmacological activities in pre-clinical trials, such as antioxidants, antileshimaniasis, anticonvulsant and wound healing.

Medicinal Potential of Garcinia Species and Their Compounds.

Garcinia is a genus of Clusiaceae, distributed throughout tropical Asia, Africa, New Caledonia, Polynesia, and Brazil. Garcinia plants contain a broad range of biologically active metabolites which, in the last few decades, have received considerable attention due to the chemical compositions of their extracts, with compounds which have been shown to have beneficial effects in several diseases. Our work had the objective of reviewing the benefits of five Garcinia species (G. brasiliensis, G. gardneriana, G. pedunculata, G. cambogia, and G. mangstana). These species provide a rich natural source of bioactive compounds with relevant therapeutic properties and anti-inflammatory effects, such as for the treatment of skin disorders, wounds, pain, and infections, having demonstrated antinociceptive, antioxidant, antitumoral, antifungal, anticancer, antihistaminic, antiulcerogenic, antimicrobial, antiviral, vasodilator, hypolipidemic, hepatoprotective, nephroprotective, and cardioprotective properties. This demonstrates the relevance of the genus as a rich source of compounds with valuable therapeutic properties, with potential use in the prevention and treatment of nontransmissible chronic diseases.

Polyprenylated benzophenone derivatives with a novel tetracyclo[8.3.1.03,11.05,10]tetradecane core skeleton from Clusia burle-marxii exhibited cytotoxicity against GL-15 glioblastoma-derived human cell line.

Three new polyprenylated benzophenone derivatives (1-3) were identified in the hexane extract of Clusia burle-marxii trunks, through the isolation and structural elucidation of their methyl derivatives, along with two known polyprenylated benzophenone derivatives sampsonine N (4) and obdeltifolione C (5). Burlemarxiones A (1) and B (2) show an unprecedent tetracyclo[8.3.1.03,11.05,10]tetradecane core skeleton. These compounds are a pair of ß-diketones in tautomeric equilibrium, whereas isonemorosonol (3) is the respective ß-diketone pair in tautomeric equilibrium with nemorosonol. Burlemarxione A methyl derivative (1a) and sampsonine N exhibited strong in vitro cytotoxic activity against GL-15 glioblastoma-derived human cell line.

A comparison of callus induction in 4 Garcinia species

Background: This research is intended to determine suitable types and concentrations of plant growth regulators (PGRs) to induce callus on stem and leaf sections of 4 species of the genus Garcinia, namely, Garcinia mangostana, Garcinia schomburgkiana, Garcinia cowa, and Garcinia celebica. The base medium was MS medium containing 30 g l -1 sucrose, 0.5 g l-1 polyvinylpyrrolidone (PVP), and 7 g l-1 agar, and for the different treatments, PGRs were added to the medium as follows: thidiazuron (TDZ) at concentrations of 0, 0.1, 0.5, 1, and 2 mg l-1; 6-(3- hydroxybenzylamino) purine (meta-topolin) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; 4-amino-3,5,6- trichloro-2-pyridinecarboxylic acid (picloram) at concentrations of 0, 0.5, 2.5, and 5 mg l-1; and 2,4- dichlorophenoxyacetic acid (2,4-D) at concentrations of 0, 0.5, 1, 2, and 4 mg l-1. The occurrence of callus was observed after 4 weeks. Results: A maximum of 100% and 93% of G. mangostana leaf explants formed callus in the 0.5 mg l-1 and 1 mg l-1 TDZ treatments, respectively, while 100% of G. schomburgkiana stem explants formed callus in the 1 mg l-1 TDZ treatment and 89% of G. schomburgkiana leaf explants formed callus in the 0.5 mg l-1 picloram treatment. The highest callus induction rate for G. cowa was 62% in the 1 mg l-1 TDZ treatment and for G. celebica was 56% in the 0.5 mg l-1•mT-1 treatment. Conclusions: For all 4 species, the greatest amount of large nodular callus was observed in the TDZ treatments. White, friable callus was observed on most of the 2,4-D and picloram treatment groups. Most meta-topolin treatments resulted in minimal callus formation.

Pollination efficiency of cockroaches and other floral visitors of Clusia blattophila.

Cockroaches have rarely been documented as pollinators. In this paper we examine whether this is because they might be inefficient at pollination compared to other pollinators. Clusia blattophila, a dioecious shrub growing on isolated rocky outcrops in French Guiana, is pollinated by Amazonina platystylata cockroaches and provides a valuable system for the study of cockroach pollination efficiency. We examined the species composition of the visitor guild and visitation rates by means of camcorder recordings and visitor sampling. Then, we investigated the capacity for pollen transfer of principal visitors and found correlations between visitation rates and pollen loads on stigmas. In an exclusion experiment we determined the contributions of individual species to pollination success. Amazonina platystylata, crickets and two species of Diptera transferred pollen, but the number of transferred pollen grains was only related to visitation rates in the case of cockroaches. Crickets visited and rarely carried pollen. Dipterans were as frequent as cockroaches, carried similar pollen loads, but transferred much less pollen. An estimated 41% and 17% of ovules were pollinated by cockroaches and dipterans, respectively. The remaining ovules were not pollinated. There was no spatial variation in pollinator guild composition, but cockroaches visited flowers less frequently at the smaller study site. We demonstrate that cockroaches pollinate a large proportion of ovules. Their pollination service is not confined to one study site and, unlike that provided by dipterans, is not limited to certain years. We suggest that cockroach pollination has been overlooked and that cockroach-pollinated plants, which share certain floral features, possess adaptations to pollination by cockroaches.

Molecular characterization of progenies of bacurizeiro ( Platonia insignis ) from Marajó Island, northeastern Amazon / Caracterização molecular de progênies de bacurizeiro ( Platonia insignis ) da Ilha de Marajó, nordeste da Amazônia

ABSTRACT The bacurizeiro (Platonia insignis Mart.) is a tree native to the Amazon whose fruit is much used in the gastronomy in the North and Northeast regions of Brazil. Due to its great economic potential for these regions, the species is being conserved in germplasm banks to support genetic breeding programs. The aim of this work was the molecular characterization of P. insignis accessions belonging to the germplasm bank of the Embrapa Eastern Amazon research unit using ISSR (Inter Simple Sequence Repeat) markers. Seventy-eight accessions of P. insignis belonging to 16 progenies were sampled in two different localities on Marajó Island, state of Pará, Brazil. Among the 16 progenies, seven were collected in Soure and nine in Salvaterra. The 78 accessions were genotyped with 23 pre-selected primers. We obtained 121 amplified products, of which 54 were polymorphic. The most polymorphic primers were UBC 834, UBC 899 and UBC 900. Primers UBC810 and UBC884 did not amplify polymorphic bands. Forty-nine markers out of 54 were selected for genetic analyses. AMOVA within and among progenies showed low genetic differentiation (PT = 0.064, P 0.001) with higher diversity within progenies (96%), low genetic differentiation among sampling localities (PT = 0.025, P 0.013), and higher diversity within (98%) than among localities. Clustering by UPGMA based on Jaccard similarities among pairs of accessions did not separate genotypes according to progeny or sampling localitiy. We recommend that new germplasm surveys consider a greater sampling effort within sampling localitites.(AU)

RESUMO O bacurizeiro (Platonia insignis Mart.) é uma espécie frutífera nativa da Amazônia muito utilizada na cultura alimentar nas regiões Norte e Nordeste do Brasil. Devido a seu grande potencial econômico regional, a espécie vem sendo conservada em bancos ativos de germoplasma (BAG) para apoiar programas de melhoramento genético. Dessa forma, o objetivo deste trabalho foi caracterizar molecularmente acessos de P. insignis pertencentes ao BAG da Embrapa Amazônia Oriental por meio de marcadores ISSR (Inter Simple Sequence Repeat). Foram coletados 78 acessos de P. insignis pertencentes a 16 progênies coletadas em dois locais diferentes na Ilha de Marajó, PA. Das 16 progênies, sete foram coletadas em Soure e nove em Salvaterra. Os 78 acessos foram genotipados com 23 primers ISSR pré-selecionados. Obteve-se 121 produtos amplificados, dos quais 54 foram polimórficos. Os primers mais polimórficos foram UBC 834, UBC 899 e UBC 900. Já os primers UBC810 e UBC884 não apresentaram bandas polimórficas. Das 54 marcas, 49 foram selecionadas para as análises genéticas. A AMOVA entre e dentro de progênies identificou baixa diferenciação genética (PT = 0,064, P 0,001) com maior diversidade dentro de progênies (96%), bem como baixa diferenciação genética entre os locais de coleta (PT = 0,025, P 0,013), com maior diversidade dentro (98%) do que entre locais. O agrupamento pelo método UPGMA, com base nas similaridades de Jaccard entre os acessos, não separou as amostras por progênie ou local de coleta. Recomenda-se que novas coletas de germoplasma considerem maior esforço de coleta em cada local amostrado.(AU)

Molecular characterization of progenies of bacurizeiro ( Platonia insignis ) from Marajó Island, northeastern Amazon / Caracterização molecular de progênies de bacurizeiro ( Platonia insignis ) da Ilha de Marajó, nordeste da Amazônia

ABSTRACT The bacurizeiro (Platonia insignis Mart.) is a tree native to the Amazon whose fruit is much used in the gastronomy in the North and Northeast regions of Brazil. Due to its great economic potential for these regions, the species is being conserved in germplasm banks to support genetic breeding programs. The aim of this work was the molecular characterization of P. insignis accessions belonging to the germplasm bank of the Embrapa Eastern Amazon research unit using ISSR (Inter Simple Sequence Repeat) markers. Seventy-eight accessions of P. insignis belonging to 16 progenies were sampled in two different localities on Marajó Island, state of Pará, Brazil. Among the 16 progenies, seven were collected in Soure and nine in Salvaterra. The 78 accessions were genotyped with 23 pre-selected primers. We obtained 121 amplified products, of which 54 were polymorphic. The most polymorphic primers were UBC 834, UBC 899 and UBC 900. Primers UBC810 and UBC884 did not amplify polymorphic bands. Forty-nine markers out of 54 were selected for genetic analyses. AMOVA within and among progenies showed low genetic differentiation (ΦPT = 0.064, P<0.001) with higher diversity within progenies (96%), low genetic differentiation among sampling localities (ΦPT = 0.025, P<0.013), and higher diversity within (98%) than among localities. Clustering by UPGMA based on Jaccard similarities among pairs of accessions did not separate genotypes according to progeny or sampling localitiy. We recommend that new germplasm surveys consider a greater sampling effort within sampling localitites.

RESUMO O bacurizeiro (Platonia insignis Mart.) é uma espécie frutífera nativa da Amazônia muito utilizada na cultura alimentar nas regiões Norte e Nordeste do Brasil. Devido a seu grande potencial econômico regional, a espécie vem sendo conservada em bancos ativos de germoplasma (BAG) para apoiar programas de melhoramento genético. Dessa forma, o objetivo deste trabalho foi caracterizar molecularmente acessos de P. insignis pertencentes ao BAG da Embrapa Amazônia Oriental por meio de marcadores ISSR (Inter Simple Sequence Repeat). Foram coletados 78 acessos de P. insignis pertencentes a 16 progênies coletadas em dois locais diferentes na Ilha de Marajó, PA. Das 16 progênies, sete foram coletadas em Soure e nove em Salvaterra. Os 78 acessos foram genotipados com 23 primers ISSR pré-selecionados. Obteve-se 121 produtos amplificados, dos quais 54 foram polimórficos. Os primers mais polimórficos foram UBC 834, UBC 899 e UBC 900. Já os primers UBC810 e UBC884 não apresentaram bandas polimórficas. Das 54 marcas, 49 foram selecionadas para as análises genéticas. A AMOVA entre e dentro de progênies identificou baixa diferenciação genética (ΦPT = 0,064, P<0,001) com maior diversidade dentro de progênies (96%), bem como baixa diferenciação genética entre os locais de coleta (ΦPT = 0,025, P<0,013), com maior diversidade dentro (98%) do que entre locais. O agrupamento pelo método UPGMA, com base nas similaridades de Jaccard entre os acessos, não separou as amostras por progênie ou local de coleta. Recomenda-se que novas coletas de germoplasma considerem maior esforço de coleta em cada local amostrado.

Clusia criuva Cambess. (Clusiaceae): anatomical characterization, chemical prospecting and antioxidant activity

ABSTRACT This study aims the anatomical description and chemical characterization of aerial parts of Clusia criuva Cambess., Clusiaceae in addition to the evaluation of the antioxidant activity of crude extracts, correlated to the flavonoid content. The morphological characterization was performed using traditional techniques of plant anatomy. For phytochemical studies, crude extracts were obtained by static maceration and analyzed by thin layer chromatography. The antioxidant activity and the flavonoids content were determined by colorimetric methods involving, respectively, 2,2-diphenyl-1-picrylhydrazyl free radical and aluminum chloride. C. criuva has uniseriate epidermis, paracytic stomata, hypostomatic leaves, cuticular flanges and cordiform vascular cylinder with accessory bundles. Chemical prospecting confirmed the abundant presence of terpenes and phenols in the extracts of leaves and of fruits. The methanolic extract of seeds showed the lowest EC50 value, but the methanolic extract of pericarps exhibited the highest maximum antioxidant activity. The results suggested a high percentage of flavonoids in the hexanic extract of pericarps, however, this could represent, in fact, the presence of benzophenones. Secretory ducts and the shape of the midrib are diagnostic for C. criuva. The antioxidant activity is not directly related to the flavonoids. The results indicate the importance of future studies with C. criuva chemical constituents.

Pharmacobotanical study of Hypericum thymopsis

ABSTRACT Hypericum thymopsis Boiss., Hypericaceae, is an endemic herb which generally grows at the calcareous steppe regions of Central Turkey. In flowering stage, the aerial parts of this species are used for wound-healing and sedation, and its infusions are used against stomach diseases and throat infections by local people. The aim of this study, to examine and to reveal of the morphology, anatomy and histology of the aerial vegetative and reproductive organs of the H. thymopsis, which are used in popular medicine and thus contributing to the pharmacognostic evaluation of the species. In comparison with previous published morphological description of the species, some different findings about plant height and leaf length were found. In addition, some morphological characteristics such as dimensions of sepals and ovaries, length of filaments, anthers, pistils and pedicels were examined here for the first time. The anatomical characteristics of stem, leaf, sepal, filament and pistil were studied using light microscopy and additionally for stem and leaf using scanning electron microscopy. The stem has the secondary growth, and circular shape. The leaves are amphistomatic and the mesophyll is dorsiventral. Stomata are anisocytic and sunken. The glandiferous emergences are present on stem and translucent glands exist in leaf mesophyll. Type A secretory canals are present in stem, leaf, sepal and petal cross section. While type B secretory canals are observed in sepal and petal, type C secretory canals are observed solely in ovary cross section. All the structural features herein found can assist the diagnosis of H. thymopsis.

Development of genomic tools in a widespread tropical tree, Symphonia globulifera L.f.: a new low-coverage draft genome, SNP and SSR markers.

Population genetic studies in tropical plants are often challenging because of limited information on taxonomy, phylogenetic relationships and distribution ranges, scarce genomic information and logistic challenges in sampling. We describe a strategy to develop robust and widely applicable genetic markers based on a modest development of genomic resources in the ancient tropical tree species Symphonia globulifera L.f. (Clusiaceae), a keystone species in African and Neotropical rainforests. We provide the first low-coverage (11X) fragmented draft genome sequenced on an individual from Cameroon, covering 1.027 Gbp or 67.5% of the estimated genome size. Annotation of 565 scaffolds (7.57 Mbp) resulted in the prediction of 1046 putative genes (231 of them containing a complete open reading frame) and 1523 exact simple sequence repeats (SSRs, microsatellites). Aligning a published transcriptome of a French Guiana population against this draft genome produced 923 high-quality single nucleotide polymorphisms. We also preselected genic SSRs in silico that were conserved and polymorphic across a wide geographical range, thus reducing marker development tests on rare DNA samples. Of 23 SSRs tested, 19 amplified and 18 were successfully genotyped in four S. globulifera populations from South America (Brazil and French Guiana) and Africa (Cameroon and São Tomé island, FST  = 0.34). Most loci showed only population-specific deviations from Hardy-Weinberg proportions, pointing to local population effects (e.g. null alleles). The described genomic resources are valuable for evolutionary studies in Symphonia and for comparative studies in plants. The methods are especially interesting for widespread tropical or endangered taxa with limited DNA availability.

Micropropagation of Calophyllum brasiliense (Cambess. ) from nodal segments / Micropropagação de Calophyllum brasiliense (Cambess. ) a partir de segmentos nodais

Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and -naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM -naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.(AU)

A micropropagação de Calophyllum brasiliense Cambess. (Clusiaceae) é uma maneira de superar dificuldades para sua produção em larga escala, devido à natureza recalcitrante das sementes, frutificação irregular e ausência de propagação vegetativa natural da espécie. Culturas foram estabelecidas utilizando segmentos nodais com 2 cm de comprimento, obtidos de plantas com 1 a 2 anos de idade, mantidas em casa de vegetação. Cloreto de mercúrio e Plant Preservative Mixture foram utilizados durante a etapa de desinfestação, com melhores resultados alcançados com a incorporação de Plant Preservative Mixture ao meio de cultura. Polivinilpirrolidona, carvão ativado, cisteína, ácido ascórbico ou ácido cítrico foram adicionados ao meio de cultura para evitar a oxidação dos explantes. Após 30 dias de cultivo, o uso de polivinilpirrolidona ou ácido ascórbico proporcionou melhores resultados, eliminando a oxidação na maioria dos explantes. Para multiplicação das brotações, benzilaminopurina foi usada em concentrações de 4.4 e 8.8 µM em meio WPM, resultando em uma média de 4.43 e 4.68 brotações por explante, respectivamente, após 90 dias. Ácido indol-3-butírico e ácido -naftaleno acético foram usados para a indução de raízes, alcançando um enraizamento máximo de 24% com o uso de 20µM de ácido -naftaleno acético. As plantas enraizadas foram transferidas para substrato Plantmax® e cultivadas em casa de vegetação, alcançando 79% de sobrevivência após 30 dias e 60% após um ano.(AU)

Micropropagation of Calophyllum brasiliense (Cambess) from nodal segments / Micropropagação de Calophyllum brasiliense (Cambess) a partir de segmentos nodais

Abstract Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture™ were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture™ incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and α-naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM α-naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

Resumo A micropropagação de Calophyllum brasiliense Cambess. (Clusiaceae) é uma maneira de superar dificuldades para sua produção em larga escala, devido à natureza recalcitrante das sementes, frutificação irregular e ausência de propagação vegetativa natural da espécie. Culturas foram estabelecidas utilizando segmentos nodais com 2 cm de comprimento, obtidos de plantas com 1 a 2 anos de idade, mantidas em casa de vegetação. Cloreto de mercúrio e Plant Preservative Mixture™ foram utilizados durante a etapa de desinfestação, com melhores resultados alcançados com a incorporação de Plant Preservative Mixture™ ao meio de cultura. Polivinilpirrolidona, carvão ativado, cisteína, ácido ascórbico ou ácido cítrico foram adicionados ao meio de cultura para evitar a oxidação dos explantes. Após 30 dias de cultivo, o uso de polivinilpirrolidona ou ácido ascórbico proporcionou melhores resultados, eliminando a oxidação na maioria dos explantes. Para multiplicação das brotações, benzilaminopurina foi usada em concentrações de 4.4 e 8.8 µM em meio WPM, resultando em uma média de 4.43 e 4.68 brotações por explante, respectivamente, após 90 dias. Ácido indol-3-butírico e ácido α-naftaleno acético foram usados para a indução de raízes, alcançando um enraizamento máximo de 24% com o uso de 20µM de ácido α-naftaleno acético. As plantas enraizadas foram transferidas para substrato Plantmax® e cultivadas em casa de vegetação, alcançando 79% de sobrevivência após 30 dias e 60% após um ano.

Immunomodulatory and toxicological evaluation of the fruit seeds from Platonia insignis, a native species from Brazilian Amazon Rainforest

Abstract The “bacuri” (Platonia insignis Mart., Clusiaceae) is a native tropical fruit from the Brazilian Amazon and Northeast Regions. Its seeds are used to treat inflammatory diseases, diarrhea and skin problems in traditional medical practices. Regarding its widespread medicinal uses, it is important to evaluate the biological and toxicological potential of this species. This way, the aim of this study was to investigate the in vitro cytotoxic and immunomodulatory effects of the hexanic extract of P. insignis seeds, as well as its in vivo acute oral toxicity. The biological evaluation was performed by the determination of cytotoxic (MTT and hemolysis assay) and immunomodulatory (phagocytic capacity, lysosomal volume and nitrite production) activities of EHSB in murine peritoneal macrophages. In addition, the oral acute toxicity was evaluated using female Wistar rats treated with EHSB (2.0 g/kg), in accordance with the OECD 423 Guideline. The EHSB showed low toxicity for macrophages in the MTT test (CC50 value: 90.03 µg/ml), as well as for erythrocytes, which caused only 2.5% hemolysis at the highest concentration. A strong immunomodulatory activity was observed by a markedly increase of the NO production, phagocytic ability and lysosomal volume. On the other hand, it was not observed deaths or changes in the clinical and behavioral parameters in the toxicological evaluation. This manner, the present study contributes to the knowledge about the immunomodulatory and toxicological properties of the P. insignis. This may provide perspectives for the evaluation and development of effective and safe phytomedicines created from the Brazilian local biodiversity.

Micropropagation of Calophyllum brasiliense (Cambess.) from nodal segments

Abstract Micropropagation of Calophyllum brasiliense Cambess. (Clusiaceae) is a way to overcome difficulties in achieving large-scale plant production, given the recalcitrant nature of the seeds, irregular fructification and absence of natural vegetative propagation of the species. Cultures were established using nodal segments 2 cm in length, obtained from 1-2 year old seedlings, maintained in a greenhouse. Mercury chloride and Plant Preservative Mixture were used in the surface sterilizing stage, better results being achieved with Plant Preservative Mixture incorporation in culture medium, at any concentration. Polyvinylpyrrolidone, activated charcoal, cysteine, ascorbic acid or citric acid were added to the culture medium to avoid oxidation. After 30 days of culture, polyvinylpirrolidone and ascorbic acid gave better results, eliminating oxidation in most explants. For shoot multiplication, benzylaminopurine was used in concentrations of 4.4 and 8.8 µM in Woody Plant Medium, resulting in an average of 4.43 and 4.68 shoots per explant, respectively, after 90 days. Indole-3-butyric acid and -naphthalene acetic acid were used to induce root formation, reaching a maximum rooting rate of 24% with 20µM -naphthalene acetic acid. For acclimatization. the rooted plants were transferred to Plantmax® substrate and cultured in a greenhouse, reaching 79% of survival after 30 days and 60% after one year.

Resumo A micropropagação de Calophyllum brasiliense Cambess. (Clusiaceae) é uma maneira de superar dificuldades para sua produção em larga escala, devido à natureza recalcitrante das sementes, frutificação irregular e ausência de propagação vegetativa natural da espécie. Culturas foram estabelecidas utilizando segmentos nodais com 2 cm de comprimento, obtidos de plantas com 1 a 2 anos de idade, mantidas em casa de vegetação. Cloreto de mercúrio e Plant Preservative Mixture foram utilizados durante a etapa de desinfestação, com melhores resultados alcançados com a incorporação de Plant Preservative Mixture ao meio de cultura. Polivinilpirrolidona, carvão ativado, cisteína, ácido ascórbico ou ácido cítrico foram adicionados ao meio de cultura para evitar a oxidação dos explantes. Após 30 dias de cultivo, o uso de polivinilpirrolidona ou ácido ascórbico proporcionou melhores resultados, eliminando a oxidação na maioria dos explantes. Para multiplicação das brotações, benzilaminopurina foi usada em concentrações de 4.4 e 8.8 µM em meio WPM, resultando em uma média de 4.43 e 4.68 brotações por explante, respectivamente, após 90 dias. Ácido indol-3-butírico e ácido -naftaleno acético foram usados para a indução de raízes, alcançando um enraizamento máximo de 24% com o uso de 20µM de ácido -naftaleno acético. As plantas enraizadas foram transferidas para substrato Plantmax® e cultivadas em casa de vegetação, alcançando 79% de sobrevivência após 30 dias e 60% após um ano.