Repositório RCAAP

Abstract Campomanesia lineatifolia (gabiroba) is a native edible species found in the Amazon Rainforest. Previous studies have demonstrated the flavonoid nature of ethanolic extract from the C. lineatifolia leaves, in addition to gastroprotective activity and TNF inhibition. However, the extraction process used was long and consumed a large amount of solvent. Therefore, the objective of this study was to obtain a bioactive extract rich in phenolics, in an extractive method of simpler, faster, and lower-cost execution. The C. lineatifolia leaves were dried and crushed, and the extractions were carried out in different solvents/mixtures (ethanol, methanol, ethyl acetate, and water) under ultrasonic bath (UB), electromagnetic stirring, and continuous reflux extraction (R). The extraction efficiency was evaluated by the flavonoid major compound concentration in the extracts, in a method developed by ultra-high performance liquid chromatography (UHPLC). Injection and pattern matching tests, and UHPLC analyses coupled to ultraviolet spectrometry were conducted to identify catechin and quercitrin. It has been demonstrated that the ethanolic extraction by R and the mixture of ethanol: water (8:2) by UB represented optimized methods in obtaining the flavonoid compounds identified. Thus, the results may contribute to chemical-biological extract standardization for gastric antiulcer activity evaluation.

Abstract Duroia saccifera (Rubiaceae) occurs in the Amazon rainforest and their extracts showed antibacterial properties. To obtain greater quantities of active substances, leaf segments from in vitro D. saccifera seedlings were used as explants for calli induction; calli were multiplied via multiple subcultures, dried and extracted with hexane followed by ethyl acetate (EtOAc) and methanol (MeOH). As D. macrophylla had been reported to produce antimycobacterial substances, we assayed calli extracts against Mycobacterium tuberculosis (H37Rv strain). Calli EtOAc extract was active, with a minimal inhibitory concentration (MIC) of ≤ 25 mg mL-1, IC90of 19.5 mg mL-1 and minimal bactericidal concentration (MBC) of 200 mg mL-1. EtOAc extract was analyzed by Thin Layer Chromatography (TLC) and Nuclear Magnetic Resonance (NMR) to determine its chemical profile, and was found to be rich in terpenes. Chromatographic fractionation of the EtOAc extract yielded a mixture of two sterols, β-sitosterol and stigmasterol (in proportion of 2:1), which were identified by 1H and 13C NMR analysis. As far as we know, this is the first report of Duroia saccifera in vitro cell culture, antituberculosis activity of calli extract and β-sitosterol and stigmasterol isolation from in vitro plant cell culture.

Abstract Ipomoea pes-caprae (Convolvulaceae), popularly known as “salsa-da-praia”, is used in folk medicine for the treatment of several diseases, including hypertension. The aim of this research was to evaluate hypotensive activity attributed to this species and identify the bioactive compounds responsible for this effect. Hydroalcoholic extract of aerial parts of Ipomoea pes-caprae (PIEA) was fractionated by column chromatography with Amberlite XAD-2, thus achieving a methanolic fraction (PIEA-M). The chemical profile investigation of PIEA-M was conducted by Ultra Performance Liquid Chromatography coupled to Mass Spectrometry (UPLC-MS) and compound isolation through High Performance Countercurrent Chromatography (HPCCC). Hemodynamic evaluations of PIEA, PIEA-M and isolated compounds were carried out on normotensive rats. The locomotor activity of PIEA and PIEA-M were performed by Rota Rod test using Swiss mice. UPLC-MS analysis of PIEA-M led to the identification of 11 compounds corresponding to seven phenolic acids and four flavonoids. By means of HPCCC, quercetin 6”-O-acetyl-3-O- glucoside (1), a mixture of quercetin-3-O-galactoside and quercetin-3-O- glucoside (2) and quercetin 3-O-glucoside (3) were isolated. PIEA, PIEA-M and the obtained compounds (1 and 2) significantly reduced blood pressure. Also, PIEA and PIEA-M caused significant impairment on mice locomotor activity. These results indicate the potential of Ipomoea pes-caprae as a source of hypotensive agents suggesting that phenolic compounds present in the species are responsible for its hypotensive activity.

Abstract Justicia pectoralis (Acanthaceae) is employed in folk medicine for its analgesic, anti-inflammatory and sedative effects and to treat respiratory diseases. It is known for properties of its coumarins, 1,2-benzopyrone and umbelliferone. A green, simple, fast, and inexpensive ultrasound-assisted extractive (UAE) method for extracting umbelliferone and 1,2-benzopyrone from Justicia pectoralis was optimized. Additionally, a HPLC analytical method was developed and validated for the simultaneous determination of both coumarins. The Box-Behnken design and response surface methodology were used to evaluate the UAE process. Ethanol concentration, extraction time, plant-to-solvent ratio were the independent variables studied and the coumarin content was the dependent one. The HPLC-UV/VIS method was validated in terms of recovery, linearity, accuracy, precision and robustness, proving to be valuable for the quality control of Justicia pectoralis extract and in the development of its herbal products. Results show that the optimal UAE conditions were: ethanol concentration of 15% (w/w), extraction time of 34 min and plant-to-solvent ratio of 0.1 g/mL. The predicted values of coumarin contents (22.16 µg/mL - umbelliferone and 163.86 µg/mL - benzopyrone) were determined under the optimal UAE conditions and proved that UAE is an efficient and eco-friendly extractive process for the production of aerial part extracts from Justicia pectoralis.

Abstract Maytenus s.l. is a genus of the Celastraceae family and many of its species are used in traditional medicine. Most of the substances responsible for the biological activities of plants come from their secondary metabolism, such as terpenoids and flavonoids, which have multiple functions in the plant and can occur in a similar way in plant groups. Chemophenetics analyzes the occurrence of these substances in a taxon and can infer evolutionary trends and contribute to the rational search for new drugs. Chemosystematic parameters were used to analyze the data obtained through a bibliographic survey. The species of Maytenus s.l. were separated into three groups: species with accepted names, synonymous species with the genus Monteverdia and synonymous species with the genus Gymnosporia. Due to their great occurrence and structural variety in Maytenus s.l., triterpenes were chosen as chemosystematic markers, with friedelanes and quinone methide triterpenes being the most expressive types. Studies have already demonstrated the analgesic potential of quinone methides and the antitumor potential of phenolic triterpenes and dimers of these units. Together, the three classes assist in the circumscription of Maytenus s.s. and Monteverdia, while lupanes can be used in the separation of these genera. Oleanans are representative of Gymnosporia.

Abstract With the upcoming medical Cannabis regulation, quality control methods on raw material will be required. Besides testing for contaminants and potency, there are also pharmaceutical and forensic interests in the determination of the terpene profile in different strains of Cannabis as complementary identification methods. A simple non-destructive HS-SPME GC-MS method was used to identify the terpene content in twelve Cannabis samples, four of them were of the hemp type (Harle-tsu), seven from various marihuana types and one of the intermediate type. They all were previously analyzed by HPLC to determine the potency (THC and CBD content). Spectral library matching was used to identify the terpenes compounds. Thirty terpenes compounds were detected, nine of them were present in all Cannabis samples and used to find their terpene profile: α-pinene, β-pinene, β-myrcene, D-limonene, terpinolene, linalool, caryophyllene, α-bergamotene and humulene. Three of them, caryophyllene, α-pinene and β-myrcene were found as larger components in most of samples. A principal components analyses (PCA) was performed. The four hemp type samples showed two different profiles, two samples showed caryophyllene as main component and the others two with β-myrcene as such. The marihuana type samples showed wider profiles with no clear patterns at all, which is not surprising because of the low number of samples. The simple methodology shows viable to set the terpenes profile for analyses of raw Cannabis material. Suitability for differentiation between different sorts of types needs more studies, with increasing numbers of samples.

Abstract The investigation of genetic diversity in natural populations of species that show potential for use in reforestation programs is a key step in making management decisions. However, reforestation programs with native species in Brazil are still rarely based on a genetic understanding of the seed matrices used for seedling production. This is also the case for Myrsine umbellata, a dioecious shrub within the family Primulaceae that has been used in reforestation programs in Brazil, mainly due to its high production capacity of fruits attractive to the avifauna. The goal of this study was to measure intra- and interpopulational genetic diversity in natural populations of M. umbellata in six forest remnants of the Atlantic Forest using ISSR markers. The results revealed that the intrapopulational genetic diversity was greater than the genetic diversity among the studied populations. For this reason, the cultivation of seedlings from seeds obtained in more than one population seems the most appropriate strategy for reforestation purposes. Even though the most isolated populations are also the ones with highest genetic structure, all populations of M. umbellata included in this study revealed to be an important germplasm bank conserved in situ.

Abstract The present study analyzed taxa of the family Cabombaceae occurring in the state of Ceará. Only Cabomba species was represented, with C. aquatica and C. haynesii recorded in the state. Populations occur in permanent and temporary lentic environments, such as lagoons and shores of lotic systems. This work presents taxon identification keys, morphological descriptions, illustrations, comments and geographic distribution data.

Abstract Species of Acanthaceae are predominantly associated with conserved forest environments such as Iguaçu National Park (PARNA Iguaçu), which is composed of Semideciduous Seasonal Forest and Araucaria forest. The aim of this work was to perform a floristic study of Acanthaceae of PARNA Iguaçu, with botanical illustrations, an identification key and descriptions of the species. Collections were carried out monthly from August 2015 to July 2016, in the areas of Céu Azul, Capanema and Foz do Iguaçu. The individuals collected were deposited in the UNOP herbarium and the specimens present in the herbaria EVB, HCF, MBM and UNOP, as well as in the virtual herbaria Reflora and SpeciesLink, were analyzed. A total of 13 native species were recorded from Atlantic Forest, distributed in seven genera. Justicia was the most representative, with five species. In all, 12 new records were made for PARNA Iguaçu, of which eight are new records for Semideciduous Seasonal Forest. Of the species found, three are threatened with extinction, one of which is considered vulnerable and two are categorized as endangered, which reinforces the role of PARNA Iguaçu in in-situ conservation in the state of Paraná.

Abstract Three new taxa of subtribe Hyptidinae (Lamiaceae) from Amazonian Brazil are here described: two from Pará state Eriope macrostachya var. amazonica, and Hyptis cachimboensis, and Hyptis spathulata from Rondônia state. The diagnostic characters, which distinguish them from their closest relatives are described and discussed, their conservation status is given and a plate is provided for each of the new taxa. The Hyptidinae conspectus an earlier checklist of Lamiaceae from Amazonian Brazil is updated with various name changes since its publication, and with the addition of another two species of Hyptis not previously included. Also included, is a note on Hyptis sect. Eriosphaeria, to which the two new species described here, belong.

Abstract In this research we studied root anatomy of eight species and two morphotypes from the genus Geonoma in Venezuela, in order to explore their structural and taxonomic implications. Fresh material sampled wild population was fixed in FAA and hand-sectioned or with a rotary microtome, following standardized protocols. Histochemical tests were carried out in some components. Obtained preparations were studied under a calibrated optical microscope; some variables were quantified by means of an eyepiece micrometer. The obtained results allowed the anatomical differentiation of the studied species and the two morphotypes. Most informative diagnostic variables identified include the thickening shape of the cells in the exodermis walls (transverse section), parenchyma cells with circular contours in the cortex, length of air spaces, the differentiation of passage cells in the endodermis, shared “V” and “Y” shaped xylem arches, the shape of phloem strands and scattered metaxylem vessels in the pith. Our results are also discussed in the context of other structural studies published on roots of different members of Arecaceae.

Resumo Calophyllaceae está representada na Reserva Ducke por dez espécies pertencentes aos seguintes gêneros: Calophyllum (C. brasiliense), Caraipa (C. costata, C. grandifolia subsp. grandifolia, C. heterocarpa, C. odorata, C. punctulata, C. rodriguesii, C. valioi), Clusiella (C. axillaris) e Haploclathra (H. paniculata). Apresentamos descrições para os gêneros e espécies, chaves de identificação e comentários sobre distribuição geográfica e fenologia.

Resumo Lytoneuron está circunscrito a família Pteridaceae (Cheilanthoideae) e foi segregado do gênero Doryopteris s.l. a partir da evidência molecular e morfológica, reconhecido como uma linhagem monofilética. Este estudo se baseia na análise de espécimes ocorrentes no estado do Rio de Janeiro, obtidos no campo e que estão nos acervos dos herbários. Como resultado, são indicadas 11 espécies de Lytoneuron para o estado das quais nove são consideradas endêmicas do Brasil e restritas ao bioma Mata Atlântica (Lytoneuron acutilobum, L. itatiaiense, L. feei, L. ornithopus, L. paradoxum, L. quinquelobatum, L. rosenstockii, L. subsimplex e L. tijucanum). As espécies Lytoneuron quinquelobatum, L. subsimplex e L. tijucanum são conhecidas apenas para o estado Rio de Janeiro. São apresentadas descrições, comentários, ilustrações e uma chave de identificação.

Resumo Esse estudo teve como objetivo realizar o levantamento florístico das espécies de Convolvulaceae ocorrentes na Área de Proteção Ambiental Jenipabu - APAJ. Os dados foram obtidos a partir de expedições de campo aleatórias, no período de agosto de 2010 a julho de 2011. Foram registradas nove espécies, distribuídas em cinco gêneros: Daustinia montana, Distimake aegyptius, Evolvulus frankenioides, E. ovatus, Ipomoea asarifolia, I. brasiliana, I. pes-caprae, I. rosea, e Jacquemontia bahiensis. Destaca-se que D. montana, I. asarifolia e I. pes-caprae ocorreram preferencialmente em áreas úmidas, e as demais foram encontradas em áreas secas. Daustinia montana, I. brasiliana, e J. bahiensis são endêmicas do Brasil e I. rosea é endêmica do nordeste brasileiro. A morfologia das folhas, sépalas e gineceu foram os principais caracteres para diagnosticar as espécies. O tratamento possui chave de identificação, descrições, pranchas ilustrativas e comentários sobre a distribuição dos táxons.

Abstract The present study was carried out in Limpopo National Park (LNP) with the objective to map floristic communities that occur inside the park. Three (3) Landsat 8 satellite images were obtained by Operational Land Imager (OLI) and Thermal Infrared Sensor (TIRS) tools and were pre-processed and classified, culminating in six (6) types of land use and cover. The floristic survey consisted of stratified sampling in three (3) main LNP landscapes types, namely landscape of limestone soils, landscape of rhyolite shallow soils and landscape of deep sandy soils. The final map presents 13 floristic communities identified by the names of greatest value of ecological importance (IVI) species. The community of Terminalia sericea / Combretum apiculatum / Guibourtia conjugata / Colophospermum mopane presented a larger occurrence area. Colophospermum mopane was the most representative species among the mapped communities, mostly occurring in association with other species. The overall accuracy was 74% and the Kappa index was 68%, thus giving a good rating. The mapping also showed that human occupied areas are larger than some floristic communities areas, so we suggest that these smaller communities, should be given priority actions for their conservation, especially those without human occupation.

Abstract The use of micropropagation techniques is crucial for the conservation of endangered moss species and their reestablishment in nature. This study aimed to establish in vitro cultures of gametophyte fragments of ten species of Cerrado mosses. After disinfestation with alcohol and commercial bleach, moss explants were grown in Petri dishes containing Knop medium. The species Bryum argenteum, B. coronatum, Isopterygium tenerifolium, Leucobryum crispum, Pogonatum pensilvanicum, and Vitalia cuspidifera were successively established with efficiency rate ranging from 1 to 31.2%. However, no aseptic cultures were obtained for the species Barbula indica, Bryum densifolium, Fissidens flaccidus, and Sphagnum platyphylloides. Even though, a few contaminated explants of these species were able to develop and grow. Thus, all ten species were submitted to rescue techniques to establish cultures in aseptic conditions, from partially contaminated explants (indirect establishment). Consequently, the indirect establishment resulted in higher percentages of explant development, which enhanced the establishment of in vitro cultures for most of the species tested. This fact is especially important for conservation purposes, mainly for species whose material is sensitive or scarce. Therefore, indirect establishment as a new in vitro culture methodology was a viable form of propagating the bryophyte species listed in this research. This fact is essential for conservation purpose, especially for species whose material is sensitive or scarcer.

Abstract We compared the functional diversity of community at edge and interior areas of an Atlantic forest fragment to test the hypothesis that higher functional diversity exists along edges - in consonance with their higher abundance and floristic diversity as compared to the interior of the forest. By considering a set of vegetative, reproductive and phenological traits and ecosystem service aspects of edge and interior environments, we defined plant functional groups using Cluster Analysis, followed by a silhouette width analysis, together with functional diversity indices of richness, divergence, evenness and dispersion. The main functional groups formed were similar between the edges and interior. Functional richness was the only index that demonstrated differences between edge and the interior. Alterations were perceived in relation to species richness and the Shannon index. Edge effects were not significant in the formation of functional groups. In contrast to our original hypothesis, similar groups were formed both along the edge in the interior - indicating that species played similar ecological roles in both environments, with similar responses to different environmental factors - so that forest edges were colonized by a series of different species that maintained diversity patterns similar to those found in the forest interior.

Abstract In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.

Resumo Os agricultores familiares que vivem em áreas de assentamentos rurais estão entre os diversos grupos sociais da Amazônia que detêm um amplo conhecimento sobre o uso de plantas medicinais. Estudos realizados neste contexto revelam uma vasta heterogeneidade no conhecimento e uso de plantas medicinais, bem como a influência dessas comunidades na diversidade vegetal e nos aspectos culturais associados ao seu uso. Esta pesquisa traz informações sobre a dinâmica do conhecimento e uso de plantas medicinais no Assentamento Rural Paulo Fonteles, estabelecido desde o ano de 2006 no Distrito de Mosqueiro, Município de Belém, estado do Pará. Os dados foram obtidos através de entrevistas semiestruturadas e oficinas participativas e analisados qualitativa e quantitativamente. As plantas medicinais são recursos prioritários para os assentados em períodos que antecedem e sucedem a ocupação. Foram registradas 140 plantas medicinais, das quais, 119 foram identificadas e pertencem a 58 famílias botânicas. Sessenta espécies são nativas do Brasil, sendo 21 com domínio fitogeográfico restrito a Amazônia e 59 são introduzidas. O conhecimento local de plantas medicinais encontra-se bem difundido entre informantes de diferentes classes de idade e gênero. As pessoas provenientes de outras regiões do Brasil reportaram um acervo de plantas medicinais significativamente maior do que àquelas naturais do estado do Pará. Entre as 140 plantas mencionadas, 110 referem-se a espécies cujos usos foram mantidos ao longo do tempo pelos informantes. A heterogeneidade sociocultural e a dinâmica do modo de vida dos assentados rurais contribuem para o acúmulo de saberes e no incremento da diversidade da flora local.

Abstract Gymnopilus is characterized by its ferruginous-yellow basidiomata and lamellae, ferruginous spore print, ellipsoidal basidiospores with warty and rough ornamentation, and lacking a germinative pore. Here, novel data on the Gymnopilus species of Paraguay is presented, macro and microscopic morphological characteristics, distribution, and ecology are described, and a taxonomic discussion is provided. Gymnopilus imperialis is recorded in the Alto Paraná Department, G. lepidotus in the Central Department, G. luteofolius in the Cordillera Department, G. peliolepis in the Paraguarí Department, and G. purpureosquamulosus in the Central Department and Boquerón, all as new records for Paraguay. Photographs of the fresh basidiomata and some microscopic structures such as basidia and basidiospores are attached.