Antioxidant and in vitro cytogenotoxic properties of Amburana cearensis (Allemão) A.C.Sm. leaf extract.

Amburana cearensis leaves have been used in folk medicine to treat respiratory diseases and inflammations. This study aimed to evaluate the biological potential of A. cearensis leaves by antioxidant and in vitro cytogenotoxic analyses of ethanolic crude extract (EE) and its fractions in healthy human cells. The EE was obtained by percolation, followed by fractionation using dichloromethane, cyclohexane, ethyl acetate (EtOAc), and methanol (MeOH) as organic solvents. Extract and all fractions were evaluated for their antioxidant potential by DPPH and reducing power tests. In vitro cytotoxic activity was determined in human peripheral blood mononuclear cells by MTT assay for the extract, EtOAc and MeOH fractions. In turn, the genotoxic activity was determined in human lymphocytes by the Cytokinesis Block Micronucleus assay only for the EtOAc fraction. Only EtOAc fraction was analyzed via gas chromatography coupled to mass spectrometry due to its higher biological activity. Considering the antioxidant potential, the EtOAc fraction was most effective in DPPH (EC50 43.37 µg/mL) and reducing power (EC50 89.80 µg/mL) assays. GC-MS analysis of the EtOAc fraction led to the identification of guaiacol, 2,3-dihydro-benzofuran, 2-methoxy-4-vinylphenol, isovanillic acid methyl ester, 4-hydroxybenzaldehyde, and 4-(ethoxymethyl)-phenol. The EE (400-1000 µg/mL), EtOAc (≤150 µg/mL) and MeOH (50 and 150-600 µg/mL) fractions were not cytotoxic by MTT test. Additionally, the EtOAc fraction (100-400 µg/mL) did not induce significant genotoxic damage. Concentrations of the EtOAc fraction with antioxidant activity showed no cytotoxicity, nor genotoxicity potential, indicating them as a nontoxic natural antioxidant source.

Cyperus prophyllatus: An endangered aquatic new species of Cyperus L. (Cyperaceae) with a exceptional spikelet disarticulation pattern among about 950 species, including molecular phylogenetic, anatomical and (micro)morphological data.

Cyperus prophyllatus, an endangered new species of Cyperus (Cyperaceae) from an aquatic ecosystem of the Atlantic Forest, Espírito Santo State, southeastern Brazil, is described and illustrated. The spikelet morphology of Cyperus prophyllatus is unique among the c. 950 species of Cyperus in having both a conspicuous spikelet prophyll and a corky rachilla articulation, which remain persistent at the base of the spikelet after disarticulation. Our molecular phylogenetic data support the placement of C. prophyllatus in the C3 Cyperus Grade and more precisely in the clade representing Cyperus sect. Oxycaryum, which also includes C. blepharoleptos and C. gardneri. Anatomical and (micro)morphological analyses corroborate the phylogenetic results, provide a better understanding of ecology and taxonomy, as well as reveal compatibility of structures with survival and dispersion in aquatic environments. A distribution map, table with distinctive characters of allied species, and conservation status are made available.

Flora of the Usina São José, Igarassu, Pernambuco State, Brazil: Acanthaceae, Gesneriaceae and Loganiaceae / Flora da Usina São José, Igarassu, Estado de Pernambuco, Brasil: Acanthaceae, Gesneriaceae e Loganiaceae

Acanthaceae, Gesneriaceae, and Loganiaceae are well-represented in Brazil and in the Neotropical region, although they have been poorly studied in northeastern Brazil, especially in the Atlantic Forest of Pernambuco State. Thus, aiming to contribute with the taxonomic knowledge about these taxa, this study had the objective of identifying and characterizing the species found in forest fragments of the Usina São José, in Igarassu, as a continuation of the monograph series for the area. Fieldwork was carried out between January 2017 and January 2018. Specimens from the herbaria CEPEC, HST, IPA, JPB, NY, PEUFR, RB, and UFP were also analyzed. Eleven species were found in the study area, five belonging to Acanthaceae, two to Gesneriaceae, and four to Loganiaceae. An identification key, descriptions, illustrations and comments on distribution and habitats are provided.

Acanthaceae, Gesneriaceae e Loganiaceae são bem representadas na flora Neotropical e brasileira, no entanto são famílias pouco estudadas no Nordeste do país, em especial na Mata Atlântica do Estado de Pernambuco. Dessa forma, visando contribuir com o conhecimento taxonômico acerca desses táxons, este estudo teve como objetivo identificar e caracterizar morfologicamente as espécies ocorrentes nos fragmentos da Usina São José, localizada em Igarassu, dando continuidade à série de monografias para a área. Expedições de coletas foram realizadas na área entre janeiro de 2017 e janeiro de 2018. Os espécimes depositados nas coleções dos herbários CEPEC, HST, IPA, JPB, NY, PEUFR, RB e UFP também foram analisados. Para a área de estudo, foram encontradas onze espécies, cinco de Acanthaceae, duas de Gesneriaceae e quatro de Loganiaceae. São apresentados chave de identificação, descrições, ilustrações e comentários sobre distribuição geográfica e hábitats.

Advanced understanding of phylogenetic relationships, morphological evolution and biogeographic history of the mega-diverse plant genus Myrcia and its relatives (Myrtaceae: Myrteae).

Myrcia is the largest exclusively Neotropical genus of the plant family Myrtaceae with c. 770 species. Although several studies have elucidated the relationships within particular sections of the genus, to date no phylogeny has been produced that includes a broad taxonomic and geographic representation. Here we present a phylogenetic hypothesis of Myrcia and close relatives comprising 253 species and based on two nuclear and seven plastid markers. We combine previously available sequence data with 234 new sequences of the genus Myrcia for this study. We use this phylogeny to investigate the evolution of selected morphological traits and to infer the biogeographic history of the genus. Our results yield a highly supported phylogenetic tree where the Myrceugenia group is sister to the Myrcia and Plinia groups. Five Myrcia species previously considered unplaced emerge in a newly circumscribed clade. The monophyly of two Myrcia sections previously considered uncertain, Aulomyrcia and Gomidesia, are confirmed with strong support. Flowers with free calyx lobes, 2-locular ovaries, and anthers with symmetrical thecae are ancestral features of Myrcia. The Myrcia sect. Gomidesia is highly supported and recovered as monophyletic, with asymmetric anthers that retain their curvature after dehiscence as a morphological synapomorphy. The Atlantic Forest is the most likely ancestral area of the genus and most of its internal clades, from where multiple lineages colonized different regions of South and Central America, in particular the Brazilian Cerrado through multiple unidirectional range expansions. The southern Atlantic Forest is the ancestral area for Myrcia sect. Gomidesia, with lineages reaching the northern Atlantic Forest, Cerrado, Yungas, and other savanna vegetation of South America. Our results provide a solid backbone for further evolutionary and taxonomic work and clarify several previously uncertain relationships in this mega-diverse plant group, and shed light on its geographical range evolution.

Integrative research identifies 71 new plant species records in the state of Rio Grande do Norte (Brazil) and enhances a small herbarium collection during a funding shortage.

A National Forest Inventory (NFI) encompassing the entire territory of Brazil is in progress. It is coordinated and promoted by the Brazilian Forest Service of the Ministry of Environment. In each state, the NFI collaborates with local herbaria by receiving collected plant material and performing species identification. Consultants are hired by the NFI and work at the local herbaria under the supervision of a curator. In exchange for curatorial assistance, the NFI provides equipment and consumables for the herbarium. Other public projects collaborating with NFI are Reflora and the Brazilian Biodiversity Information System (SiBBr). Both projects have online platforms that seek to connect herbaria and make all their data freely available, including high quality digital images of specimens. Through inter-institutional collaboration, the joint interests of NFI, Reflora, SiBBr and local herbaria have improved collections, expanded the online Reflora database, and provided the NFI with verified species lists. These strategic uses of public funding are positively affecting Botany, particularly during a period of economic crisis and cuts in research. Here, we illustrate the increase in floristic knowledge through the improvement of a herbarium collection in Rio Grande do Norte (RN) - the Brazilian state with the lowest levels of plant richness. We report 71 new occurrences of vascular plants for RN, belonging mainly to the Poaceae, Fabaceae and Malvaceae. Most of the species with new occurrences have a Neotropical distribution (21 spp.) and only seven are restricted to the Brazilian Northeast. Our findings highlight previous gaps in RN's floristic knowledge. The partnership NFI, Reflora, SiBBr and the UFRN herbarium improved herbarium curation, digital collection, and quality of data. Finally, a fellowship provided by Reflora and SiBBr allowed improving curation by distributing duplicates and incorporating the Herbarium of Câmara Cascudo Museum.

Population genetic structure of the rock outcrop species Encholirium spectabile (Bromeliaceae): The role of pollination vs. seed dispersal and evolutionary implications.

PREMISE OF THE STUDY: Inselbergs are terrestrial, island-like rock outcrop environments that present a highly adapted flora. The epilithic bromeliad Encholirium spectabile is a dominant species on inselbergs in the Caatinga of northeastern Brazil. We conducted a population genetic analysis to test whether the substantial phenotypic diversity of E. spectabile could be explained by limited gene flow among populations and to assess the relative impact of pollen vs. seed dispersal on the genetic structure of the species. METHODS: Nuclear and chloroplast microsatellite markers were used to genotype E. spectabile individuals from 20 rock outcrop locations, representing four geographic regions: northern Espinhaço Range, Borborema Plateau, southwestern Caatinga and southeastern Caatinga. F-statistics, structure, and other tools were applied to evaluate the genetic makeup of populations. KEY RESULTS: Considerable levels of genetic diversity were revealed. Genetic structuring among populations was stronger on the plastid as compared with the nuclear level, indicating higher gene flow via bat pollination as compared with seed dispersal by wind. structure and AMOVA analyses of the nuclear data suggested a high genetic differentiation between two groups, one containing all populations from the southeastern Caatinga and the other one comprising all remaining samples. CONCLUSIONS: The strong genetic differentiation between southeastern Caatinga and the remaining regions may indicate the occurrence of a cryptic species in E. spectabile. The unique genetic composition of each inselberg population suggests in situ conservation as the most appropriate protection measure for this plant lineage.

Inside-out flowers of Lacandonia brasiliana (Triuridaceae) provide new insights into fundamental aspects of floral patterning.

Background and Aims. A recently described Brazilian species, Lacandonia brasiliana, shares with its longer established putative sister species from Mexico, L. schismatica, inverted floral patterning (carpels surrounding stamens) that is almost unique among angiosperms. We present a detailed ontogenetic study of L. brasiliana for comparison with other members of the tribe Triurideae (Triuridaceae) to explore the possible evolutionary origins of "inside-out" flowers. Methods. Wild-source populations of L. brasiliana were compared morphologically and ontogenetically with related species of Triurideae, using light and scanning electron microscopy. Key Results. Relatively few morphological differences separate flowers of L. brasiliana and L. schismatica. Both species have tepals with late-developing subapical appendages. In both species, the three central (almost sessile) anthers develop precociously with respect to the carpels; the anthers remain closed, and fertilization is achieved via pollen-tube growth from germinating pollen grains of the same cleistogamous flower. Carpels are initiated on fascicles. Conclusions. The close similarity between the two Lacandonia species makes it unlikely that they arose independently from two separate homeotic transformation events; they could either represent sister species or two populations of a single disjunct species. Our study underlines the problematic generic and species boundaries within Triurideae. We present an evolutionary scenario of character evolution in Triuridaceae. The inside-out Lacandonia flower could have resulted from a stabilized homeotic transformation; this hypothesis is not in conflict with constrasting theories of the origin of the Triuridaceae flower, which coincided with a shift to unisexuality. The unisexual yet highly plastic flowers that are typical of Triuridaceae could have pre-adapted the origin of the extraordinary Lacandonia morphology.

Unraveling the karyotype structure of the spurges Euphorbia hirta Linnaeus, 1753 and E. hyssopifolia Linnaeus, 1753 (Euphorbiaceae) using genome size estimation and heterochromatin differentiation.

Euphorbia Linnaeus, 1753 (Euphorbiaceae) is one of the most diverse and complex genera among the angiosperms, showing a huge diversity in morphologic traits and ecologic patterns. In order to improve the knowledge of the karyotype organization of Euphorbia hirta (2n = 18) and Euphorbia hyssopifolia (2n = 12), cytogenetic studies were performed by means of conventional staining with Giemsa, genome size estimations with flow cytometry, heterochromatin differentiation with chromomycin A3 (CMA) and 4',6-diamidino-2-phenylindole (DAPI) and Giemsa C-banding, fluorescent in situ hybridization (FISH) with 45S and 5S rDNA probes, and impregnation with silver nitrate (AgNO3). Our results revealed small metacentric chromosomes, CMA+/DAPI0 heterochromatin in the pericentromeric regions of all chromosomes and CMA+/DAPI- in the distal part of chromosome arms carriers of nucleolar organizing regions (NORs). The DNA content measurements revealed small genomes for both species: Euphorbia hirta with 2C = 0.77 pg and Euphorbia hyssopifolia with 2C = 1.41 pg. After FISH procedures, Euphorbia hirta, and Euphorbia hyssopifolia presented three and four pairs of terminal 45S rDNA sites, respectively, colocalizing with CMA+ heterochromatic blocks, besides only one interstitial pair of 5S rDNA signals. Additionally, the maximum number of active NORs agreed with the total number of observed 45S rDNA sites. This work represents the first analysis using FISH in the subfamily Euphorbioideae, revealing a significant number of chromosomal markers, which may be very helpful to understand evolutionary patterns among Euphorbia species.

Occurrence and evolutionary inferences about Kranz anatomy in Cyperaceae (Poales).

Cyperaceae is an angiosperm family with the greatest diversity of species with Kranz anatomy. Four different types of Kranz anatomy (chlorocyperoid, eleocharoid, fimbristyloid and rhynchosporoid) have been described for this angiosperm family, and the occurrence and structural characteristics of these types are important to trace evolutionary hypotheses. The purpose of this study was to examine the available data on Cyperaceae Kranz anatomy, emphasizing taxonomy, geographic distribution, habitat and anatomy, to infer the potential origin of the Kranz anatomy in this family. The results showed that the four types of Kranz anatomy (associated with C4 photosynthesis) in Cyperaceae emerged numerous times in unrelated phylogenetic groups. However, the convergence of these anatomical types, except rhynchosporoid, was observed in certain groups. Thus, the diverse origin of these species might result from different environmental pressures that promote photorespiration. Greater variation in occurrence of Kranz anatomy and anatomical types was observed in Eleocharis, whose emergence of the C4 pathway was recent compared with other genera in the family, and the species of this genus are located in aquatic environments.

DNA barcoding in Atlantic Forest plants: What is the best marker for Sapotaceae species identification?

The Atlantic Forest is a phytogeographic domain with a high rate of endemism and large species diversity. The Sapotaceae is a botanical family for which species identification in the Atlantic Forest is difficult. An approach that facilitates species identification in the Sapotaceae is urgently needed because this family includes threatened species and valuable timber species. In this context, DNA barcoding could provide an important tool for identifying species in the Atlantic Forest. In this work, we evaluated four plant barcode markers (matK, rbcL, trnH-psbA and the nuclear ribosomal internal transcribed spacer region - ITS) in 80 samples from 26 species of Sapotaceae that occur in the Atlantic Forest. ITS yielded the highest average interspecific distance (0.122), followed by trnH-psbA (0.019), matK (0.008) and rbcL (0.002). For species discrimination, ITS provided the best results, followed by matK, trnH-psbA and rbcL. Furthermore, the combined analysis of two, three or four markers did not result in higher rates of discrimination than obtained with ITS alone. These results indicate that the ITS region is the best option for molecular identification of Sapotaceae species from the Atlantic Forest.

Importance of anatomical leaf features for characterization of three species of Mapania (Mapanioideae, Cyperaceae) from the Amazon Forest, Brazil / Importância dos caracteres anatômicos foliares na caracterização de três espécies de Mapania (Mapanioideae, Cyperaceae) da Floresta Amazônica, Brasil

Mapania belongs to Mapanioideae, a quite controversial subfamily in Cyperaceae due to the existence of unusual characters in both reproductive and vegetative organs. The genus is represented by seven species in Northern Brazil but taxonomic valuable information related to the leaf organs is still unknown. The present study aimed the anatomical description of the leaf organs (either basal leaves or cataphylls and involucral bracts) of three representative Brazilian species of Mapania. Samples of cataphylls, basal leaves and involucral bracts were sectioned and stained for observations under light microscopy. The involucral bracts provide the most elucidative characters (ten) to distinguish the three species The basal leaves provides six distinguishing characters and are useful to M. macrophylla and M. pycnostachya, as they are absent in M. sylvatica. Mesophyll arrangement in the involucral bracts supports the circumscription of M. macrophylla and M. pycnostachya in M. sect. Pycnocephala and of M. sylvatica in M. sect. Mapania. Some features as thin-walled epidermal cells, stomata level and aerenchyma were considered to be adaptive to the humid environment in which the species occur. The translucent cells are here considered as aerenchyma precursors and a supportive function is assumed for the bulliform cells on the basal leaves and involucral bracts. No silica bodies were found which confirm it as a diagnostic character of Mapania among Hypolytreae genera.

Mapania pertence à subfamília Mapanioideae que apresenta caracteres incomuns às demais Cyperaceae, tanto vegetativos como reprodutivos. O gênero é representado por sete espécies no norte do Brasil e apresenta ainda lacunas de informações especialmente relacionadas aos órgãos foliares. O presente estudo objetivou a descrição anatômica dos órgãos foliares (folhas basais, ou catafilos, e brácteas involucrais) de três espécies representativas de Mapania no território brasileiro. Amostras de catafilos, folhas basais e brácteas involucrais foram seccionadas e coradas para observações em microscopia de luz. As brácteas involucrais foram as mais elucidativas na separação das espécies, com dez caracteres variáveis, uma vez que ocorrem em todas as espécies durante o estágio reprodutivo. As folhas basais forneceram seis caracteres úteis na distinção de M. macrophylla e M. pycnostachya, já que não ocorrem em M. sylvatica. seguidas das folhas basais. O arranjo do mesofilo das brácteas involucrais suporta a circunscrição de M. macrophylla e M. pycnostachya em M. sect. Pycnocephala e de M. sylvatica em M. sect. Mapania. Características adaptativas ao ambiente úmido também foram encontradas, como epiderme com paredes delgadas, níveis dos estômatos e presença de aerênquima. As células translúcidas do mesofilo foram descritas como precursoras de aerênquima e considerou-se que as células buliformes presentes nas folhas e brácteas involucrais apresentam função de suporte desses órgãos. Corpos silicosos não foram encontrados sustentando o seu uso na delimitação de Mapania e dos demais gêneros de Hypolytreae.