Unlocking the Complete Chloroplast Genome of a Native Tree Species from the Amazon Basin, Capirona (Calycophyllum Spruceanum, Rubiaceae), and Its Comparative Analysis with Other Ixoroideae Species.

Capirona (Calycophyllum spruceanum Benth.) belongs to subfamily Ixoroideae, one of the major lineages in the Rubiaceae family, and is an important timber tree. It originated in the Amazon Basin and has widespread distribution in Bolivia, Peru, Colombia, and Brazil. In this study, we obtained the first complete chloroplast (cp) genome of capirona from the department of Madre de Dios located in the Peruvian Amazon. High-quality genomic DNA was used to construct libraries. Pair-end clean reads were obtained by PE 150 library and the Illumina HiSeq 2500 platform. The complete cp genome of C. spruceanum has a 154,480 bp in length with typical quadripartite structure, containing a large single copy (LSC) region (84,813 bp) and a small single-copy (SSC) region (18,101 bp), separated by two inverted repeat (IR) regions (25,783 bp). The annotation of C. spruceanum cp genome predicted 87 protein-coding genes (CDS), 8 ribosomal RNA (rRNA) genes, 37 transfer RNA (tRNA) genes, and one pseudogene. A total of 41 simple sequence repeats (SSR) of this cp genome were divided into mononucleotides (29), dinucleotides (5), trinucleotides (3), and tetranucleotides (4). Most of these repeats were distributed in the noncoding regions. Whole chloroplast genome comparison with the other six Ixoroideae species revealed that the small single copy and large single copy regions showed more divergence than inverted regions. Finally, phylogenetic analyses resolved that C. spruceanum is a sister species to Emmenopterys henryi and confirms its position within the subfamily Ixoroideae. This study reports for the first time the genome organization, gene content, and structural features of the chloroplast genome of C. spruceanum, providing valuable information for genetic and evolutionary studies in the genus Calycophyllum and beyond.

Phytochemical and morpho-anatomical study of the vegetative organs of Psychotria fractistipula L. B.Sm., R.M. Klein & Delprete (Rubiaceae)

Psychotria fractistipula L.B.Sm, R.M. Klein & Delprete (Rubiaceae) is found in the existing Atlantic Forest in the South of Brazil, in the Paraná and Santa Catarina States. The members of Rubiaceae are characterized chemically by the presence of alkaloids, some pharmacological properties of which include antifungal, antibacterial, analgesic, antioxidant, antimutagenic, and antiviral activities. In this study, we report the results of anatomical, morphological, and histochemical analyses of the leaves and stems of the Rubiaceae family member, P. fractistipula. The anatomical analysis involved of light microscopy and scanning electron microscopy. Morpho-anatomical and histochemical characterization were made using standard methodology. The isolated compounds were identified by Nuclear Magnetic Resonance spectroscopic analysis. The following structures are highlighted as the distinctive features of the species: presence of petiolate leaves and a petiole base, leaf blade with smooth cuticle on both surfaces, paracytic stomata, and, dorsiventral mesophyll and presence of trichomes on the leaves and stems. Phytosteroids, identified as β-sitosterol and campesterol, were observed in the hexane fraction of the stem. Future studies should focus on the isolation of other fractions of interest and their characterization through specific biological and cytotoxicity assays.

The monotypic Brazilian genus Diacrodon is a synonym of Borreria (Spermacoceae, Rubiaceae): morphological and molecular evidences.

Diacrodon is a monotypic genus of the tribe Spermacoceae (Rubiaceae), endemic to northeastern Brazil. Diacrodon compressus is frequently misidentified with a two lobed calyx species of Borreria, B. verticillata. Traditionally, in Spermacoceae the fruit type was considered a diagnostic character among the genera. In this sense, D. compressus presents a strongly compressed, one seeded and indehiscent fruit (vs. globose, two seeded and dehiscent fruit in B. verticillata). In this work, we address two objectives: evaluate the systematic position and determine the identity of Diacrodon in respect to other taxa. Molecular analyses using ITS and ETS indicate that D. compressus is strongly related to other species of Borreria. The morphological results revealed that D. compressus, despite of its type of fruit, is identical to Borreria in floral and palynological features. As conclusion, the new combination Borreria diacrodonta is made and a lectotype is designated. An updated description of the species and a key to the Borreria species with a two lobed calyx are provided. The distribution of B. diacrodonta is extended to Brazilian states Goiás and Minas Gerais, and Paraguay. By this taxonomical change it has become clear that the dehiscence of the fruits lack taxonomic value in the delimitation of Borreria.

Rubiaceae in Brazilian Atlantic Forest remnants: floristic similarity and implications for conservation / Rubiaceae en remanentes de Bosque Atlántico de Brasil: similitud florística e implicaciones para la conservación

AbstractBrazil holds most of the Atlantic Forest Domain and is also one of the Rubiaceae diversity centers in the Neotropics. Despite the urban expansion in the state of Rio de Janeiro, large areas of continuous vegetation with high connectivity degree can still be found. Recently, new Rubiaceae species have been described in the Rio de Janeiro flora, which present small populations and very particular distribution. The current paper analyzed the similarity in the floristic composition of the Rubiaceae in eight Atlantic Forest remnants of Rio de Janeiro state protected by Conservation Units. We also surveyed and set guidelines for conservation of microendemic species. The similarity analysis were based on previously published studies in Área de Proteção Ambiental de Grumari, Área de Proteção Ambiental Palmares, Parque Estadual da Serra da Tiririca, Parque Nacional do Itatiaia, Parque Nacional de Jurubatiba, Reserva Biológica de Poço das Antas, Reserva Biológica do Tinguá and Reserva Ecológica de Macaé de Cima - using the PAST software (“Paleontological Statistics”) with Sørensen coefficient. The floristic similarity analysis revealed two groups with distinct physiographic characteristics and different vegetation types. Group A consisted in two Restinga areas, Área de Proteção Ambiental de Grumari and Parque Nacional de Jurubatiba, which showed strong bootstrap support (98 %). Group B included forest remnants with distinct phytophisiognomies or altitudes, but with moderate bootstrap support. Low similarity levels among the eight areas were found due to the habitats’ heterogeneity. The current study pointed out 19 microendemic species from the Atlantic Forest, they present a single-site distribution or a distribution restricted to Mountain and Metropolitan regions of Rio de Janeiro state. Concerning the conservation status of microendemic species, discrepancies between the Catalogue of Flora of Rio de Janeiro and the Red Book of Brazilian Flora (two of the main reference catalogs of Brazilian flora) have been identified. We have also highlighted the need for recollecting microendemic species from the Atlantic Forest, and for properly assessing the degree of threat faced by these taxons early. Rev. Biol. Trop. 64 (2): 655-665. Epub 2016 June 01.

ResumenBrasil tiene la mayor parte de bosque del Atlántico y también es uno de los centros de diversidad de Rubiaceas del Neotrópico. A pesar de la expansión urbana en el estado de Río de Janeiro, todavía se pueden encontrar grandes áreas de vegetación con alto grado de conectividad. Recientemente, nuevas especies de Rubiaceae se han descrito en la flora de Río de Janeiro, las cuales presentan poblaciones pequeñas y distribuciones particulares. El presente documento analiza la similitud en la composición florística de Rubiaceae en ocho remanentes del bosque Atlántico del estado de Río de Janeiro, protegidos por las Unidades de Conservación. También se inspeccionaron y establecieron directrices para la conservación de especies microendémicas. El análisis de similitud se basó en estudios publicados anteriormente en el área de Protección Ambiental de Grumari, área de Protección Ambiental Palmares, Parque Estatal de Serra da Tiririca, Parque Nacional de Itatiaia, Parque Nacional de Jurubatiba, Reserva Biológica de Poço das Antas, Reserva Biológica de Tinguá y Reserva Ecológica de Macaé de Cima, utilizando el software PAST (“Paleontological Statistics”) con coeficiente de Sørensen. El análisis de similitud florística reveló dos grupos con características fisiográficas distintas y diferentes tipos de vegetación. El Grupo A consistió en dos áreas, Área de Protección Ambiental de Grumari y el Parque Nacional de Jurubatiba, que mostraron un fuerte bootstrap support (98 %). El Grupo B incluye remanentes de bosque con fitofisionomía o altitudes distintas, pero con moderado bootstrap support. Los bajos niveles de similitud entre las ocho áreas fueron encontrados debido a la heterogeneidad de los hábitats. El estudio señaló que 19 especies microendémicas del Atlántico presentan una distribución de un solo sitio o una distribución restringida a las regiones de montaña y regiones metropolitanas del estado de Río de Janeiro. En cuanto al estado de conservación de especies microendémicas, se han identificado discrepancias entre el Catálogo de la Flora de Río de Janeiro y el Libro Rojo de la Flora de Brasil (dos de los principales catálogos de referencias de la flora brasileña). También se puso de relieve la necesidad de recolectar especies microendémicas del bosque Atlántico para evaluar temprano y adecuadamente el grado de amenaza que enfrentan estos taxones.

Rubiaceae in Brazilian Atlantic Forest remnants: floristic similarity and implications for conservation.

Brazil holds most of the Atlantic Forest Domain and is also one of the Rubiaceae diversity centers in the Neotropics. Despite the urban expansion in the state of Rio de Janeiro, large areas of continuous vegetation with high connectivity degree can still be found. Recently, new Rubiaceae species have been described in the Rio de Janeiro flora, which present small populations and very particular distribution. The current paper analyzed the similarity in the floristic composition of the Rubiaceae in eight Atlantic Forest remnants of Rio de Janeiro state protected by Conservation Units. We also surveyed and set guidelines for conservation of microendemic species. The similarity analysis were based on previously published studies in Área de Proteção Ambiental de Grumari, Área de Proteção Ambiental Palmares, Parque Estadual da Serra da Tiririca, Parque Nacional do Itatiaia, Parque Nacional de Jurubatiba, Reserva Biológica de Poço das Antas, Reserva Biológica do Tinguá and Reserva Ecológica de Macaé de Cima - using the PAST software ("Paleontological Statistics") with Sørensen coefficient. The floristic similarity analysis revealed two groups with distinct physiographic characteristics and different vegetation types. Group A consisted in two Restinga areas, Área de Proteção Ambiental de Grumari and Parque Nacional de Jurubatiba, which showed strong bootstrap support (98 %). Group B included forest remnants with distinct phytophisiognomies or altitudes, but with moderate bootstrap support. Low similarity levels among the eight areas were found due to the habitats' heterogeneity. The current study pointed out 19 microendemic species from the Atlantic Forest, they present a single-site distribution or a distribution restricted to Mountain and Metropolitan regions of Rio de Janeiro state. Concerning the conservation status of microendemic species, discrepancies between the Catalogue of Flora of Rio de Janeiro and the Red Book of Brazilian Flora (two of the main reference catalogs of Brazilian flora) have been identified. We have also highlighted the need for recollecting microendemic species from the Atlantic Forest, and for properly assessing the degree of threat faced by these taxons early.

Determination of toxicity in rabbits and corresponding detection of monofluoroacetate in four Palicourea (Rubiaceae) species from the Amazonas state, Brazil.

Numerous monofluoroacetate (MFA)-containing plants in Brazil cause sudden death syndrome precipitated by exercise in livestock, which is characterized by loss of balance, ataxia, labored breathing, muscle tremors, and recumbence leading to death. Four species of Palicourea collected at six farms were tested for the presence of MFA and their toxicity to rabbits. Palicourea longiflora and Palicourea barraensis contained MFA and caused sudden death in the rabbits. Palicourea croceoides and Palicourea nitidella did not contain MFA and were not toxic to rabbits. P. longiflora and P. barraensis were collected at three farms with a history of sudden death in their cattle. This is the first report of toxicity in regard to these two species.

Sobre a botânica, a etnofarmacologia e a química de Calycophyllum spruceanum (Benth. ) Hook. f. ex K. Schum / On the botany, ethnopharmacology, and chemistry of Calycophyllum spruceanum (Benth. ) Hook. f. ex K. Schum

RESUMO O fato de possuir espécies com potencial econômico ilimitado, faz com que o ambiente amazônico se torne alvo constante da biopirataria e da extração predatória dos recursos. Muitas espécies deste ambiente sequer possuem catalogação e, mesmo assim, encontram risco eminente de desaparecimento. Neste âmbito, trabalhos que reúnam dados sobre estas espécies possuem grande valor científico, cultural e econômico, sendo este o estímulo que desencadeou a produção desta revisão. Aqui a espécie Calycophyllum spruceanum (Benth.) Hook. f. ex K. Schum., uma Rubiaceae amazônica, é contemplada por meio de características botânicas, indicações etnofarmacológicas e propriedades químicas. Popularmente conhecida como mulateiro, a espécie é frequentemente receitada na etnomedicina como cicatrizante e rejuvenescedor, além de ser usada no controle de manchas de pele. Como peculiaridade botânica, mulateiro apresenta tronco retilíneo com epiderme fina esverdeada que evolui para uma periderme castanho-escuro, que é anualmente renovada. Em termos químicos, há destaque para a presença de alcalóides, taninos e, sobretudo, secoiridóides (7-metoxididerrosideo, 6´-acetil-β-D-glucopiranosildiderrosideo e 8-0-tigloildiderrosideo são peculiares à espécie). Mesmo com propriedades fotoprotetoras comprovadas, C. spruceanum ainda carece de pesquisas, sobretudo àquelas voltadas para a produção ex situ da planta e àquelas que demonstrem a relação entre a ecologia da planta e a produção de metabólitos funcionais para a indústria.

ABSTRACT The Amazonian environment is a constant target for biopiracy and predatory extraction of resources due to the species with vast economic potential. Many of these species have not even been cataloged, and yet are already at imminent risk of extinction. In this paper, Calycophyllum spruceanum (Benth.) Hook. f. ex k. Schum., an Amazonian Rubiaceae, is studied with information on its botanical characteristics, ethnopharmacological uses, and chemical properties. Popularly known as mulateiro, the species is often prescribed in ethnomedicine for healing and vitality, and is also used to control skin patches. As a botanical peculiarity, it has a thin green epidermis that evolves into a dark brown periderm, which is renewed annually. In terms of chemistry, there is emphasis on the presence of alkaloids, tannins, and, especially, secoiridoids (7-methoxydiderroside, 6′-acetyl-β-D-glucopyranosyldiderroside, and 8-O-tigloyldiderroside are peculiar to the species). Even with proven photoprotective properties, research on C. spruceanum is still lacking, in particular studies aimed at the ex situ production of the plant and those that show the relationship between the plant’s ecology and the production of functional metabolites for the industry.

Atividades biológicas de espécies amazônicas de Rubiaceae / Biological activities of Amazonian species of Rubiaceae

RESUMORubiaceae é uma família botânica com grande potencial químico e biológico Neste trabalho realizou-se a prospecção fitoquímica e avaliou-se as atividades citotóxica, antioxidante e antifúngica dos extratos diclorometânicos e metanólicos das folhas de Duroia saccifera, Ferdinandusa goudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera e P. guianensis. A avaliação da citotoxicidade foi realizada empregando-se o ensaio de toxicidade sobre Artemia salina. A atividade antifúngica foi determinada pelo método de difusão em ágar utilizando-se os fungos: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T. rubrum ATCC 28189, T. tonsurans 21/97, Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 e C. albicans U. 5/99. A atividade antioxidante foi determinada através dos ensaios de redução do radical livre 1,1-difenil-2-picrilhidrazila (DPPH) e de descoloração do cátion radical ácido-6-sulfônico-2,2-azinobis-3-etilbenzotiazolina (ABTS). Para o ensaio de toxicidade sobre A. salina observou-se maior toxicidade no extrato metanólico de F. goudotiana, que se mostrou tóxico até a concentração de 5 µg mL-1. A presença de princípios antifúngicos foi observada em F. hirsuta e F. paraensiscontra quatro dos fungos testados. Na avaliação da atividade antioxidante os extratos metanólicos mostraram maior atividade que os extratos diclorometânicos, sendo coincidente o resultado obtido através de ambos os métodos (redução do DPPH e descoloração do ABTS). Os resultados criam possibilidades para futuras investigações relacionadas à estrutura e atividade dos componentes de cada extrato ativo.

ABSTRACTRubiaceae is a plant family with great chemical and biological potential. In this work a phytochemical prospection was performed and it was assessed the cytotoxic, antioxidant and antifungal activities of dichloromethanic and methanolic extracts of the leaves of Duroia saccifera, Ferdinandusagoudotiana, F. hirsuta, F. paraensis, Ferdinandusa sp., Palicourea corymbifera and P. guianensis. The cytotoxicity evaluation was carried out using the test toxicity on Artemia salina. The antifungal activity was determined by agar diffusion method using fungi: Epidermophyton floccosum, Microsporum canis 32905, M. gipseum 29/00, Trichophyton mentagrophytes ATCC 9533/03, T.rubrum ATCC 28189, T. tonsurans 21/97 , Cladosporium cladosporioides, C. sphaerospermum, FusariumU. 662/06, Scytalidium U. 661/06, Candida albicans ATCC 3632 and C. albicans U. 5/99. The antioxidant activity was determined by testing the amount of free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the discoloration of the radical cation 6-sulfonic acid-2 ,2-azinobis-3-ethylbenzothiazoline (ABTS). For the toxicity test on A. salina greater toxicity in the methanol extract of F. goudotiana was observed, which proved to be toxic up to a concentration of 5 µg mL-1. The presence of antifungal principles was observed in the F. hirsuta and F. paraensis against four tested fungi. In the evaluation of the antioxidant activity, the methanol extracts showed greater activity than the dichloromethanic ones, being similar the obtained result through both methods (reduction of DDPH and ABTS decolorization) . The results create possibilities for future researches related to the structure and activity of the components of each active extract.

Cinética de secagem e difusividade efetiva em folhas de jenipapo (Genipa americana L. ) / Drying kinetics and effective diffusivity in jenipapo sheets (Genipa americana L. )

RESUMO O jenipapo (Genipa americana L.) é uma espécie nativa com importância medicinal, sendo amplamente utilizada no Brasil. Em função da necessidade de conhecimento à cerca do pré-processamento desta espécie, este trabalho foi desenvolvido com o objetivo de avaliar a cinética de secagem de folhas de jenipapo (G. americana L.), bem como determinar a difusividade efetiva da água durante o processo. As folhas foram coletadas com teor de água inicial de 2,30±0,05 (decimal b.s.), e submetidas à secagem em três condições de temperatura do ar (35,3; 46,0 e 65,0°C) até atingirem o teor de água de equilíbrio. Aos dados experimentais, ajustaram-se doze modelos matemáticos, recomendados para representar o processo de secagem de produtos agrícolas. As magnitudes do coeficiente de determinação (R2), do erro médio relativo (P), do erro médio estimado (SE) e do teste do qui-quadrado (χ2), foram utilizadas para verificar o grau de ajuste dos modelos. Os modelos de Henderson e Pabis modificado e Midilli apresentaram ajustes adequados aos dados experimentais, sendo o modelo de Midilli, em função de sua simplicidade, escolhido para representar a cinética de secagem das folhas de jenipapo. Aumentando a temperatura do ar de secagem de 35,3 para 46,0 e 65,0ºC houve redução no tempo de secagem das folhas de jenipapo de 91,1 para 62,5 e 24,2 horas, respectivamente. O coeficiente de difusão efetivo aumenta com a elevação da temperatura, e esta relação é descrita pela equação de Arrhenius, que apresenta energia de ativação para a difusão líquida de 33,9 kJ mol-1.

ABSTRACT The jenipapo (Genipa americana L.) is a native species with medicinal importance and is widely used in Brazil. Due to the need for knowledge about the preprocessing of this species, this work was developed to evaluate the drying kinetics of the jenipapo leaves (G. americana L.), and also to determine the effective diffusivity of water during the process. The leaves were collected with an initial moisture content of 2.30 ± 0.05 (decimal db), and dried in three conditions of air temperature (35.3, 46.0 and 65.0°C) until they reach the equilibrium moisture content. The experimental data set were twelve mathematical models, recommended to represent the drying process of agricultural products. The magnitudes of the coefficient of determination (R2), the mean relative error (P), the average estimated error (SE) and the chi-square (X2), were used in order to verify the adequacy level of the models. The Henderson, modified Pabis and Midilli models presented appropriate adjustments to the experimental data, with the model Midilli, due to its simplicity, chosen to represent the drying kinetics of the jenipapo leaves. By increasing the temperature of the drying air from 35.3 to 46.0 and 65.0ºC, there was a reduction in the drying time of the jenipapo leaves, from 91.1 to 62.5 and to 24.2 hours, respectively. The effective diffusion coefficient increases with the temperature’s raise, and this relationship is described by the Arrhenius equation, which shows activation energy for liquid diffusion of 33.9 kJ mol-1.

Detection of toxic monofluoroacetate in Palicourea species.

Numerous plant species worldwide including some Palicourea (Rubiaceae), Tanaecium (Bignoniaceae), and Amorimia (Malpighiaceae) species in Brazil cause sudden death and are known to contain monofluoroacetate (MFA). Two species of Palicourea, Palicourea aenofusca and Palicourea marcgravii, cause sudden death and are reported to contain MFA while other Palicourea species are reported to cause sudden death in livestock and are suspected to contain MFA due to the similarity in clinical signs. Using an HPLC-APCI-MS method to detect MFA, herbarium specimens representing 46 Palicourea taxa were screened for the presence of MFA. Additionally we screened five Psychotria taxa that are closely related to Palicourea species. Ten species of Palicourea were identified that contained MFA, two previously reported and eight newly reported here; these are closely related to each other, though some other related species did not contain MFA.

Morphology, ecophysiology and germination of seeds of the neotropical tree Alibertia patinoi (Rubiaceae).

Alibertia patinoi (Rubiaceae) is of economic and cultural importance for communities in the Colombian Pacific and Amazon regions, where it is cultivated and mature fruits are highly appreciated and consumed. Since there is a lack of knowledge of the seed physiology of this species, we describe here the germination behavior and morphometry of seeds of Alibertia patinoi, and relate them to its habitat. Fruits were collected from a mixed food crop and a commercial plantation in Guaimía village, Buenaventura, Colombia, a tropical rain forest area. We measured length, width, thickness, mass (n = 1 400), and moisture content of seeds (n = 252). Primary dormancy tests were conducted (n = 200), followed by imbibition (n=252) and germination dynamics, under different conditions of light and temperature specific to understory and forest clearings (n = 300 seeds). Finally, seed storage behavior was established (n = 100 seeds). We observed that size and mass of seeds had a narrow range of values that did not differ within or among fruits and that the species did not exhibit primary dormancy. The seeds are recalcitrant, and recently harvested seeds exhibited higher seed moisture content (ca. 44%) and continuous metabolism. The seed germination percentage was observed to be higher under the specific dense canopy forest light and temperature conditions; furthermore, neither enriched far-red light nor darkness conditions inhibited germination. We concluded that rapid germination could be the establishment strategy of this species. Also, the physiological traits (i.e., rapid germination rate, low germination requirements, absence of primary dormancy, and recalcitrant behavior) and seed size and mass, suggest that A. patinoi is adapted to conditions of mature tropical rain forests.

Morphology, ecophysiology and germination of seeds of the Neotropical tree Alibertia patinoi (Rubiaceae) / Morfología, ecofisiología y germinación de semillas del árbol neotropical

Alibertia patinoi (Rubiaceae). Alibertia patinoi (Rubiaceae) is of economic and cultural importance for communities in the Colombian Pacific and Amazon regions, where it is cultivated and mature fruits are highly appreciated and consumed. Since there is a lack of knowledge of the seed physiology of this species, we describe here the germination behavior and morphometry of seeds of Alibertia patinoi, and relate them to its habitat. Fruits were collected from a mixed food crop and a commercial plantation in Guaimía village, Buenaventura, Colombia, a tropical rain forest area. We measured length, width, thickness, mass (n=1 400), and moisture content of seeds (n=252). Primary dormancy tests were conducted (n=200), followed by imbibition (n=252) and germination dynamics, under different conditions of light and temperature specific to understory and forest clearings (n=300 seeds). Finally, seed storage behavior was established (n=100 seeds). We observed that size and mass of seeds had a narrow range of values that did not differ within or among fruits and that the species did not exhibit primary dormancy. The seeds are recalcitrant, and recently harvested seeds exhibited higher seed moisture content (ca. 44%) and continuous metabolism. The seed germination percentage was observed to be higher under the specific dense canopy forest light and temperature conditions; furthermore, neither enriched far-red light nor darkness conditions inhibited germination. We concluded that rapid germination could be the establishment strategy of this species. Also, the physiological traits (i.e., rapid germination rate, low germination requirements, absence of primary dormancy, and recalcitrant behavior) and seed size and mass, suggest that A. patinoi is adapted to conditions of mature tropical rain forests.

La germinación y morfometría de las semillas de Alibertia patinoi se describen y se relacionan con el hábitat de la especie, el Bosque Pluvial Tropical Americano. Se midió la longitud, ancho, grosor, peso y contenido de humedad de las semillas. Se realizaron pruebas de latencia, imbibición y dinámica de la germinación con diferentes condiciones de luz y temperatura específicas de claros y de sotobosque cubierto por un dosel denso. Además, se estableció el comportamiento de almacenamiento de las semillas. El tamaño y peso de las semillas no difieren dentro ni entre frutos. Las semillas recién cosechadas no tienen latencia primaria, presentan alto contenido de humedad (ca. 44%), metabolismo continuo y son recalcitrantes. La germinación tuvo porcentajes altos y en condiciones de luz y temperatura específicas de bosque con follaje denso, y ni la luz enriquecida con rojo lejano ni la oscuridad inhibieron la germinación. Se concluye que la germinación rápida podría ser la estrategia de establecimiento de esta especie. Además, los rasgos fisiológicos, el tamaño y peso de las semillas, sugieren que A. patinoi está adaptada a las condiciones del bosque lluvioso tropical maduro y se comporta como una especie que no es pionera.

Essential oil of Mitracarpus frigidus as a potent source of bioactive compounds.

In our previous work (Fabri et al. 2009), we showed that different extracts of Mitracarpus frigidus had significant antibacterial, antifungal and leishmanicidal activities. In order to increase our knowledge about this species, this work assesses the chemical composition and the in vitro biological activity of its essential oil. Thus, the essential oil obtained by hydrodistillation of the aerial parts of M. frigidus was analyzed by GC/MS. Among several compounds detected, 11 were identified, being linalool and eugenol acetate the major components. The essential oil exhibited a moderate antibacterial effect against Staphyloccocus aureus, Bacillus cereus, Pseudomonas aeruginosa and Enterobacter cloacae (MIC 250 µg/mL). On the other hand, it showed a strong antifungal effect against Cryptoccocus neoformans (MIC 8 µg/mL) and Candida albicans (MIC 63 µg/mL). Expressive activity against L. major and L. amazonensis promastigote forms with IC50 values of 47.2 and 89.7 µg/mL, respectively, were also observed. In addition, the antioxidant activity was investigated through DPPH radical-scavenging and showed a significative activity with IC50 of 38 µg/mL. The cytotoxicity against Artemia salina was moderate with LC(50) of 88 µg/mL. The results presented here are the first report on the chemical composition and biological properties of M. frigidus essential oil.

Phenology of tropical understory trees: patterns and correlates

Reproductive phenologies of plants are constrained by climate in highly seasonal regions. In contrast, plants growing in wet tropical forests are freed from many abiotic constraints, which in canopy tree communities lead to a rich diversity of phenological patterns within and among individuals, species and communities. However, basic descriptions of tropical phenological patterns and the processes that shape them are rare. Here, we document the individual-, population-, and landscape-level phenological patterns of two dominant families of understory woody plants important to avian frugivores, the Melastomataceae and Rubiaceae, along an elevational transect in Costa Rica. The 226 individual plants belonging to 35 species in this study, varied in the number of reproductive bouts/year, and the timing, duration, and synchrony of reproductive stages. This variation was not related to factors related to their interactions with mutualists and antagonists, nor did it appear to be constrained by phylogeny. Diverse phenological patterns among species led to relatively aseasonal patterns at the community and landscape level. Overall, evidence for biotic processes shaping temporal patterns of fruiting phenology was weak or absent. These findings reveal a number of unexplained patterns, and suggest that factors shaping phenology in relatively aseasonal forests operate in idiosyncratic ways at the species level.

En regiones con marcada estacionalidad, los patrones fenológicos de las plantas están limitados por el clima. Por el contrario, las plantas que crecen en bosques húmedos tropicales, no tienen tantas limitaciones abióticas y es por esto que el dosel presenta una diversidad muy rica en los patrones fenológicos de individuos, especies y comunidades. Sin embargo, es muy escasa la información sobre la descripción básica de los patrones fenológicos tropicales y de los procesos que los afectan. En este documento, presentamos los patrones fenológicos, a nivel de individuo, población y paisaje, a lo largo de un transecto altitudinal en Costa Rica, para dos familias dominantes de plantas leñosas (Melastomataceae y Rubiaceae) que son de gran importancia en la dieta de aves frugívoras. En este estudio, las plantas variaron en el número de brotes reproductivos por año, así como en el tiempo, duración y sincronización de la reproducción. Esta variación no estuvo relacionada con interacciones con mutualistas y antagonistas, ni aparentemente estuvo limitada por la filogenia. Los diversos patrones fenológicos de las especies fueron relativamente no estacionales a nivel de comunidad y paisaje. En conjunto, hubo poca evidencia de que los procesos abióticos hayan afectado los patrones fenológicos de fructificación. Estos resultados revelan una serie de patrones sin explicación aparente, y sugieren que los factores que determinan la fenología en los bosques relativamente no estacionales, funcionan de manera idiosincrática a nivel de especie.

Phenology of tropical understory trees: patterns and correlates.

Reproductive phenologies of plants are constrained by climate in highly seasonal regions. In contrast, plants growing in wet tropical forests are freed from many abiotic constraints, which in canopy tree communities lead to a rich diversity of phenological patterns within and among individuals, species and communities. However, basic descriptions of tropical phenological patterns and the processes that shape them are rare. Here, we document the individual-, population-, and landscape-level phenological patterns of two dominant families of understory woody plants important to avian frugivores, the Melastomataceae and Rubiaceae, along an elevational transect in Costa Rica. The 226 individual plants belonging to 35 species in this study, varied in the number of reproductive bouts/year, and the timing, duration, and synchrony of reproductive stages. This variation was not related to factors related to their interactions with mutualists and antagonists, nor did it appear to be constrained by phylogeny. Diverse phenological patterns among species led to relatively aseasonal patterns at the community and landscape level. Overall, evidence for biotic processes shaping temporal patterns of fruiting phenology was weak or absent. These findings reveal a number of unexplained patterns, and suggest that factors shaping phenology in relatively aseasonal forests operate in idiosyncratic ways at the species level.

Population structure of Rudgea parquioides (Rubiaceae), a shade-tolerant shrub species, in Southern Brazil

The expectations that shade-tolerant forest species show 1) a population structure composed by a high amount of small individuals, and 2) biomass allocation for diameter higher than for height growth, were tested for Rudgea parquioides, a typical shrub in Southern Brazil. We described the size structure (height and stem diameter) and allometrical relations of a R. parquioides population by counting and measuring all the individuals in a 725m² area in the municipality of Curitiba (25"25'S; 49"19'W). A total of 916 individuals (12,634 ind.ha-1) were recorded in the area. The firstexpectation was supported, since distribution by height and diameter classes showed a predominance of small individuals (skewness coefficients > 1). On the other hand, the regression between height and stem base diameter showed slope β < 1, which indicates that growth in height is higher than in diameter, not supporting the second expectation. These results show that life strategies in shade-tolerant species may imply in more trade-off combinations than previously described.

Foram testadas duas hipóteses relativas a espécies florestaistolerantes à sombra em uma população de Rudgea parquioides, um arbusto característico do sul do Brasil: 1) espécies tolerantes à sombra possuem uma estrutura populacional composta por um grande número de indivíduos pequenos e 2) a alocação de biomassa para o crescimento em diâmetro é maior que para o crescimento em altura. Foram descritas as estruturas de tamanho (altura e diâmetro do tronco) e relações alométricas através da contagem e medição de todos os indivíduos de R. parquioides em uma área de 725m² no município de Curitiba (25"25'S; 49"19'W). Um total de 916 indivíduos(12.634 ind.ha-1) foi medido nesta área. A primeira hipótese foi aceita, uma vez que a distribuição em classes de altura e de diâmetro mostrou um predomínio de indivíduos pequenos (coeficiente de assimetria > 1). Por outro lado, a regressão entre altura e diâmetro na base do caule mostrou inclinação β < 1, o que indica que o crescimento em altura é maior que em diâmetro, não suportando a segunda hipótese. Esses resultados mostram que as estratégias de crescimento em espécies tolerantes à sombra podem implicar em outras relações ecológicas além das já descritas.

Population structure of Rudgea parquioides, a shade-tolerant shrub species, in Southern Brazil.

The expectations that shade-tolerant forest species show 1) a population structure composed by a high amount of small individuals, and 2) biomass allocation for diameter higher than for height growth, were tested for Rudgea parquioides, a typical shrub in Southern Brazil. We described the size structure (height and stem diameter) and allometrical relations of a R. parquioides population by counting and measuring all the individuals in a 725m(2) area in the municipality of Curitiba (25°25'S; 49°19'W). A total of 916 individuals (12,634 ind.ha(-1)) were recorded in the area. The first expectation was supported, since distribution by height and diameter classes showed a predominance of small individuals (skewness coefficients > 1). On the other hand, the regression between height and stem base diameter showed slope ß < 1, which indicates that growth in height is higher than in diameter, not supporting the second expectation. These results show that life strategies in shade-tolerant species may imply in more trade-off combinations than previously described.

Tracing the impact of the Andean uplift on Neotropical plant evolution.

Recent phylogenetic studies have revealed the major role played by the uplift of the Andes in the extraordinary diversification of the Neotropical flora. These studies, however, have typically considered the Andean uplift as a single, time-limited event fostering the evolution of highland elements. This contrasts with geological reconstructions indicating that the uplift occurred in discrete periods from west to east and that it affected different regions at different times. We introduce an approach for integrating Andean tectonics with biogeographic reconstructions of Neotropical plants, using the coffee family (Rubiaceae) as a model group. The distribution of this family spans highland and montane habitats as well as tropical lowlands of Central and South America, thus offering a unique opportunity to study the influence of the Andean uplift on the entire Neotropical flora. Our results suggest that the Rubiaceae originated in the Paleotropics and used the boreotropical connection to reach South America. The biogeographic patterns found corroborate the existence of a long-lasting dispersal barrier between the Northern and Central Andes, the "Western Andean Portal." The uplift of the Eastern Cordillera ended this barrier, allowing dispersal of boreotropical lineages to the South, but gave rise to a huge wetland system ("Lake Pebas") in western Amazonia that prevented in situ speciation and floristic dispersal between the Andes and Amazonia for at least 6 million years. Here, we provide evidence of these events in plants.

Posible influencia de los rametes en la estructura poblacional y distribución del árbol gigante Warszewiczia uxpanapensis, Cordillera de Tilarán, Costa Rica / Possible influence of ramets on the population structure and distribution of the giant tree Warszewiczia uxpanapensis, Tilarán Mountain Range, Costa Rica

The distribution and population structure of Warszewiczia (=Elaeagia) uxpanapensis, a tree that reaches a height of 45 m, a DBH of 1.40 m and 8 m tall buttresses, was studied on two 1-ha plots of premontane pluvial forest. It had a frequency of 49 % and a mean dominance of 6.5 %. The adult distribution was random (mean distance to the nearest neighboring tree in plot 1, it was 14.26 m, n = 15; Clark-Evans Index (IC-E) = 1.07; in plot 2, it was 15.33 m, n = 18, IC-E = 1.34,). The seedlings represented 60.3 % of the sampled population (plot 1, n = 94; plot 2, n = 85). This species has a very high capability of propagating itself vegetatively through clones ("ramets"), which influence its distribution and population structure. With a ramet density in plot 1 of 0.8 and in plot 2 of 0.3/100 m2, its propagation could be from fallen trees or branches (which also create clearings in the forest) or from standing trees and even live branches that produce adventitious roots.

Se estudió los rametes, la distribución y estructura de la población de Warszewiczia uxpanapensis, en 2 ha de bosque pluvial premontano. Este árbol alcanza hasta 45 m de altura, DAPs hasta de 1.40 m y gambas de 8 m de altura. Su frecuencia fue de 49 % y su cobertura promediode 6.5 %. La distribución de los adultos fue aleatoria (distancia promedio al vecino más cercano en la parcela 1 de 14.26 m, IC-E= 1.07, n= 15 en la parcela 2 de 15.33 m, n= 18, IC-E= 1.34). Sus plántulas representaron el 60.3 % de la población estudiada (parcela 1, n= 94; parcela 2, n= 85). Se descubrió que esta especie tiene una alta capacidad de propagarse vegetativamente por rametes, los cuales influyen en la estructura de población y distribución de la especie (con una densidad de rametes parcela 1, 0.8 y parcela 2, 0.3 rametes/100 m2). Su propagación vegetativa puede ser a partir de troncos caídos o ramas desprendidas (las cuales abren claros en el bosque), rebrotes de árboles, o ramas todavía adheridas a los árboles que son capaces de producir raíces adventicias aéreas.

[Possible influence of ramets on the population structure and distribution of the giant tree Warszewiczia uxpanapensis, Tilarán Mountain Range, Costa Rica]. / Posible influencia de los rametes en la estructura poblacional y distribución del árbol gigante Warszewiczia uxpanapensis, Cordillera de Tilarán, Costa Rica.

The distribution and population structure of Warszewiczia (=Elaeagia) uxpanapensis, a tree that reaches a height of 45 m, a DBH of 1.40 m and 8 m tall buttresses, was studied on two 1-ha plots of premontane pluvial forest. It had a frequency of 49 % and a mean dominance of 6.5 %. The adult distribution was random (mean distance to the nearest neighboring tree in plot 1, it was 14.26 m, n = 15; Clark-Evans Index (IC-E) = 1.07; in plot 2, it was 15.33 m, n = 18, IC-E = 1.34,). The seedlings represented 60.3 % of the sampled population (plot 1, n = 94; plot 2, n = 85). This species has a very high capability of propagating itself vegetatively through clones ("ramets"), which influence its distribution and population structure. With a ramet density in plot 1 of 0.8 and in plot 2 of 0.3/100 m2, its propagation could be from fallen trees or branches (which also create clearings in the forest) or from standing trees and even live branches that produce adventitious roots.