Phytochemical Analysis, Antioxidant and Antifungal Activity of Essential oil and Extracts of Alpinia malaccensis (Burm. f. ) Roscoe flowers

Abstract The present study describes chemical composition, phytochemicals, antifungal activities, antioxidant assays and total phenolic content of essential oil and varied polarity solvent extract from flowers of Alpinia malaccensis (Burm.f.). Total 27 components were identified in essential oil by GC-MS with terpinen-4-ol (28.6%) and α- terpineol (12.8%) as the main constituent. The essential oil was found to have maximal levels of phenolic content (64.60 µg/mL) as compared to the other extracts. The antioxidant assay evaluated in extracts and essential oil by different methods revealed good-to-moderate antioxidant potential with different IC50 values viz. (188.02 -250.25 µg/mL) in Fe3+ reducing power, (153.15-201.59 µg/mL) in Fe2+ metal-chelating ability, (130.39-181.12 µg/mL) in DPPH, (88.29-187.32 µg/mL) in OH radical, (79.04-156.79 µg/mL), in NO radical and (138.72-233.00 µg/mL) in superoxide anion scavenging activities, respectively. The methanolic extract display remarkable fungicidal activity against the tested pathogens followed by dichloromethane extract, essential oil, hexane extract and petroleum ether extract respectively, with MIC values ranging from 31.25 to 500 µg/mL. Based on results, it can be inferred that the flower of A. malaccensis if explored further for its medicinal properties, might be a good source to develop a safe and sustainable natural food preservative

Effect of rape flower on benign prostatic hyperplasia in rats

We were carried out to investigate the efficacy of Rape (Rapeseed, Brassica napus L.) flower on BPH (benign prostatic hyperplasia) in rats. We found that the extract from Rape flower prevented hyperplasia in testosterone-induced BPH model, the relevant animal model of human BPH. Extract reduced the weight of prostate and induced significantly cell apoptosis in prostate in BPH model. In addition, the extract controlled expression of TGF-ß1 in prostate gland and promoted urinary output in dose-dependence in BPH model. Our data provide that Rape flower may be useful for treatment of BPH

Purified novel and new diferuloyl glycerate related phenolic acid from Pandanus odoratissimus flowers shows antioxidant, invertase inhibition and control against diabetic foot ulcer (DFU) causing bacterial pathogens - An in vitro study to establish an effective regulation over type 2 diabetes mellitus

Abstract Chronic type 2 diabetes mellitus (T2DM) and its associated diseases are major concern among human population and also responsible for significant mortality rate. Hence, the present study aims to evaluate and correlate the invertase inhibition, antioxidant activity and control against DFU causing bacterial pathogens by Pandanus odoratissimus flowers. Two dimensional preparative thin layer chromatography (2D PTLC) was adopted to purify the phenolic acid component and LC-MS2 was done to predict the phenolic acid structures. Standard spectrophotometry methods were adopted to investigate the in vitro invertase inhibitory and antioxidant (CUPRAC and ABTS) activities. Agar well diffusion and broth dilution assays were used to record the antibacterial property against DFU causing pathogens isolated from clinical samples. Statistical analyses were used to validate the experiments. A new and novel diferuloyl glycerate related phenolic acid (m/z 442) purified from PTLC eluate has recorded satisfactory cupric ion reducing power (ED50= 441.4±2.5 µg), moderate ABTS radical scavenging activity (IC50= 450.3±10 µg; 32.5±1.5%), and a near moderate, in vitro, invertase mixed type inhibition (24.5±4.5%; Ki: 400 µg). Similarly, bacterial growth inhibitory kinetics has showed a significant inhibition against E. coli and S. aureus.

Thrombolysis, clotting, and genotoxic activities modulated by essential oils extracted from Lippia alba

Abstract The composition and pharmacological properties of Lippia alba (Mill.) (L. alba) (Verbenaceae) flower and leaf essential oils (EO) were determined in this study. The major constituents in the flower EO were geranial (49.83%) and neral (32.75%), and in the leaf EO were geranial (38.06%), neral (31.02%), and limonene (18.03%). Flower EO inhibited thrombolysis induced by Bothrops moojeni (B. moojeni) and Lachesis muta muta (L. muta muta) venoms (0.05-1.2 µL mL-1). When tested against L. muta muta venom, the protective effect was smaller in both EO. The EOs prolonged the clotting time induced by L. muta muta venom and a procoagulant effect was observed on B. moojeni. In the comet assay, the flower EO presented anti-genotoxic action (damage frequency of only 11.6 - 34.9%) against the L. muta muta venom. The positive control (Doxorubicin) and the venom alone presented a damage frequency of 80.3% and 70.7%, respectively. The flower EO protected DNA from damage induced by L. muta muta venom. L. alba leaf and flower EOs presented anti-genotoxic action

Combination of Sanger and target-enrichment markers supports revised generic delimitation in the problematic 'Urera clade' of the nettle family (Urticaceae).

Urera Gaudich, s.l. is a pantropical genus comprising c. 35 species of trees, shrubs, and vines. It has a long history of taxonomic uncertainty, and is repeatedly recovered as polyphyletic within a poorly resolved complex of genera in the Urticeae tribe of the nettle family (Urticaceae). To provide generic delimitations concordant with evolutionary history, we use increased taxonomic and genomic sampling to investigate phylogenetic relationships among Urera and associated genera. A cost-effective two-tier genome-sampling approach provides good phylogenetic resolution by using (i) a taxon-dense sample of Sanger sequence data from two barcoding regions to recover clades of putative generic rank, and (ii) a genome-dense sample of target-enrichment data for a subset of representative species from each well-supported clade to resolve relationships among them. The results confirm the polyphyly of Urera s.l. with respect to the morphologically distinct genera Obetia, Poikilospermum and Touchardia. Afrotropic members of Urera s.l. are recovered in a clade sister to the xerophytic African shrubs Obetia; and Hawaiian ones with Touchardia, also from Hawaii. Combined with distinctive morphological differences between Neotropical and African members of Urera s.l., these results lead us to resurrect the previously synonymised name Scepocarpus Wedd. for the latter. The new species epiphet Touchardia oahuensis T.Wells & A.K. Monro is offered as a replacement name for Touchardia glabra non H.St.John, and subgenera are created within Urera s.s. to account for the two morphologically distinct Neotropical clades. This new classification minimises taxonomic and nomenclatural disruption, while more accurately reflecting evolutionary relationships within the group.

Investigation of genes associated with petal variations between diploid and autotetraploid in Chinese cabbage (Brassica rapa L. ssp. pekinensis) by RNA-seq and sRNA-seq.

Polyploidy promotes morphological, physiological, and reproductive diversity in plants. The imminent effect of chromosome doubling in plants is the enlargement of organs such as flowers and fruits, which increases the commercial value of crops. Flowering plays a vital role in the growth and development of angiosperms. Here, we prepared an isolated microspore culture of 'FT', a doubled haploid (DH) line of Chinese cabbage (Brassica rapa L. ssp. pekinensis), and obtained diploid and autotetraploid plants with the same genetic background. Compared with diploids, the autotetraploids were characterized by large floral organs, dark petals, delayed flowering, and reduced fertility. The indole-3-acetic acid (IAA) and jasmonic acid (JA) levels in autotetraploid petals were significantly higher and the abscisic acid (ABA) level was significantly lower than those in the diploid petals. The lutein level in autotetraploid petals was nearly two times higher than that in the diploid petals. A comparative transcriptome analysis revealed 14,412 differentially expressed genes (DEGs) between the diploids and autotetraploids, and they were enriched in 117 Gene Ontology terms and 110 Kyoto Encyclopedia of Genes and Genomes pathways. We detected 231 DEGs related to phytohormone signal transduction and 29 DEGs involved in carotenoid biosynthesis. An miRNA-target mRNA analysis showed that 32 DEGs regulated by 16 DEMs were associated with flowering timing (BraA03000336, BraA09004319, and BraA09000515), petal development (BraA05002408, BraA01004006, BraA09004069, and BraA04000966), flower opening (BraA07000350), and pollen development (BraA01000720, BraA09005727, and BraA01000253). This study provides information to help elucidate the molecular mechanisms underlying phenotypic variations induced by autopolyploidy in Chinese cabbage.

Taxonomic traits in the microstructure of flowers of parasitic Orobanche picridis with particular emphasis on secretory structures.

Orobanche picridis is an obligate root parasite devoid of chlorophyll in aboveground organs, which infects various Picris species. Given the high level of phenotypic variability of the species, the considerable limitation of the number of taxonomically relevant traits (mainly in terms of generative elements), and the low morphological variation between species, Orobanche is regarded as one of the taxonomically most problematic genera. This study aimed to analyse the taxonomic traits of O. picridis flowers with the use of stereoscopic and bright-field microscopy as well as fluorescence, scanning, and transmission electron microscopy. The micromorphology of sepals, petals, stamens, and pistils was described. For the first time, the anatomy of parasitic Orobanche nectaries and the ultrastructure of nectaries and glandular trichomes were presented. Special attention was paid to the distribution and types of glandular and non-glandular trichomes as well as the types of metabolites contained in these structures. It was demonstrated that the nectary gland was located at the base of the gynoecium and nectar was secreted through modified nectarostomata. The secretory parenchyma cells contained nuclei, large amyloplasts with starch granules, mitochondria, and high content of endoplasmic reticulum profiles. Nectar was transported via symplastic and apoplastic routes. The results of histochemical assays and fluorescence tests revealed the presence of four groups of metabolites, i.e. polyphenols (tannins, flavonoids), lipids (acidic and neutral lipids, essential oil, sesquiterpenes, steroids), polysaccharides (acidic and neutral polysaccharides), and alkaloids, in the trichomes located on perianth elements and stamens.

Molecular phylogenetics and floral evolution of the Cirrhopetalum alliance (Bulbophyllum, Orchidaceae): Evolutionary transitions and phylogenetic signal variation.

The Cirrhopetalum alliance is a loosely circumscribed species-rich group within the mega-diverse genus Bulbophyllum (Orchidaceae). The monophyletic status of the alliance has been challenged by previous studies, although established sectional classifications have yet to be tested in a phylogenetic context. We used maximum likelihood and Bayesian analyses of DNA sequence data (cpDNA: matK and psbA-trnH; nrDNA: ITS and Xdh; 3509 aligned characters; 117 taxa), including all sections putatively associated with the Cirrhopetalum alliance, to reconstruct the phylogeny. We mapped 11 selected categorical floral characters onto the phylogeny to identify synapomorphies and assess potential evolutionary transitions across major clades. Our results unequivocally support the recognition of an amended Cirrhopetalum alliance as a well-supported monophyletic group characterized by clear synapomorphies, following the inclusion of sect. Desmosanthes and the exclusion of five putative Cirrhopetalum-allied sections. Most sections within the Cirrhopetalum alliance are demonstrated to be polyphyletic or paraphyletic, necessitating a new sectional classification. The inclusion of sect. Desmosanthes revolutionizes our understanding of the alliance, with significant evolutionary transitions in floral characters detected. We further investigated six continuously variable characters of the sepals and labellum, and detect phylogenetic conservatism in labellum width and the evolutionary lability of lateral sepal length, which can partly be explained by the different functional roles they play in pollination and pollinator trapping.

New Caffeoylquinic Acid Derivatives and Flavanone Glycoside from the Flowers of Chrysanthemum morifolium and Their Bioactivities.

The Chrysanthemum morifolium flower is widely used in China and Japan as a food, beverage, and medicine for many diseases. In our work, two new caffeoylquinic acid derivatives (1, 2), a new flavanone glycoside (3), and six reported flavanones (4⁻9) were isolated and identified from the flowers of C. morifolium. The chemical structures of all isolates were elucidated by the analysis of comprehensive spectroscopic data as well as by comparison with previously reported data. The isolated constituents 1⁻8 were evaluated for their neuroprotective activity, and compounds 3 and 4 displayed neuroprotective effects against hydrogen peroxide-induced neurotoxicity in human neuroblastoma SH-SY5Y cells.

Style polymorphism in Linum (Linaceae): a case of Mediterranean parallel evolution?

Heterostyly is a sex polymorphism that has challenged evolutionary biologists ever since Darwin. One of the lineages where heterostyly and related stylar conditions appear more frequently is Linum (Linaceae). This group is particularly suitable for testing competing hypotheses about ancestral and transitional stages on the evolutionary building up of heterostyly. We generated a phylogeny of Linum based on extensive sampling and plastid and nuclear DNA sequences, and used it to trace the evolution of character states of style polymorphism. We also revised available data on pollination, breeding systems, and polyploidy to analyse their associations. Our results supported former phylogenetic hypotheses: the paraphyly of Linum and the non-monophyly of current taxonomic sections. Heterostyly was common in the genus, but appeared concentrated in the Mediterranean Basin and the South African Cape. Ancestral character state reconstruction failed to determine a unique state as the most probable condition for style polymorphism in the genus. In contrast, approach herkogamy was resolved as ancestral state in some clades, giving support to recent hypotheses. Some traits putatively related to heterostyly, such as life history and polyploidy, did show marginal or non-significant phylogenetic correlation, respectively. Although pollinator data are limited, we suggest that beeflies are associated with specific cases of heterostyly. The consistent association between style polymorphism and heteromorphic incompatibility points to ecological factors as drivers of the multiple evolution of style polymorphism in Linum. Albeit based on limited evidence, we hypothesised that specialised pollinators and lack of mating opportunities drive evolution of style polymorphism and loss of the polymorphism, respectively.

Bioactivity of arid region honey: an in vitro study.

BACKGROUND: Antioxidant and anti-inflammatory properties of honey have been largely recognized by various studies. Almost all of the potential benefits are associated with polyphenol content. Honey varieties from the arid region are reported to be rich in polyphenols, but data related to its bioactivity in vitro is greatly lacking. This study aimed at establishing the antioxidant and anti-inflammatory properties of arid region honey. Four honey varieties from arid region (H1, H2, H3, and H4) and two popular non-arid region honey (H5 and H6) were tested in vitro in this study. METHODS: The erythrocyte membrane protection effect of honey varieties were measured by hemolysis assay after exposing erythrocytes to a peroxide generator. The subsequent production of MDA (malondialdehyde) content in erythrocytes was measured. Immunomodulatory effect of the honey varieties was tested in prostate cancer cells PC-3 and PBMC (peripheral blood mononuclear cells) by measuring the IL-6 (interleukin 6) and NO (nitric oxide) levels in cell culture supernatant after incubation with the honey varieties. PC-3 cell viability was assessed after incubation with honey varieties for 24 h. RESULTS: Arid region honey exhibited superior erythrocyte membrane protection effect with H4 measuring 1.3 ± 0.042mMTE/g and H2 measuring 1.122 ± 0.018mMTE/g. MDA levels were significantly reduced by honey samples, especially H4 (20.819 ± 0.63 nmol/mg protein). We observed a significant decrease in cell population in PC-3 after 24 h in culture on treatment with honey. A moderate increase in NO levels was observed in both cultures after 24 h at the same time levels of IL-6 were remarkably reduced by honey varieties. CONCLUSION: The results demonstrate the antioxidant effect of arid region honey due to its erythrocyte membrane protection effect and subsequent lowering of oxidative damage as evident from lower levels of lipid peroxidation byproduct MDA. Arid region honey varieties were as good as non-arid region types at decreasing cell viability of prostate cancer cells. The moderate increase in NO levels in PC-3 and PBMCs were not significant enough to elicit any pro-inflammatory response. However, IL-6 secretion was remarkably reduced by all honey varieties in a comparable level indicating the potential anti-inflammatory property of arid region honey.

Time to split Salvia s.l. (Lamiaceae) - New insights from Old World Salvia phylogeny.

AIMS: Salvia L. is widely known as the largest genus in the mint family. A morphological modification of the androecium (lever-like stamens) was used to support this genus. However, molecular data revealed that Salvia is polyphyletic. Since phylogenetic studies largely underrepresented Old World Salvia species, we filled this gap and combined new data with existing sequences. The aim of our study was the identification of well-supported clades that provide the basis for evolutionary and taxonomic conclusions. METHODS: We included ITS data (internal transcribed spacer) from 220 Salvia species, 86 of which were sequenced for the first time. Additionally, the highly variable plastid marker rpl32-trnL was sequenced, providing new data for 100 Salvia species. These sequences were combined with the accessions available from GenBank. Old World Salvia is represented herein with 57% of its species. The two datasets were analyzed separately using BI and ML approaches. RESULTS: Our data confirm that Salvia is polyphyletic with four distinct evolutionary lineages (Clade I-IV), including five additional genera. The clades strongly reflect the geographical distribution, i.e., Clade IV (East Asia), Clade III (Southwest Asia to Northern Africa), and Clade II (America). The origin of Salvia s.s. (Clade I) is most likely Southwest Asia. A high degree of parallel character evolution was identified in most of the Old World sections. Based on our results, we reconstructed the evolution and biogeography of Salvia s.l. and propose to split this large group into six genera, each supported by geographical distribution, morphology, and karyology. CONCLUSION: Salvia s.l. is a polyphyletic group that was originally regarded as a genus because its species share a derived stamen structure. However, phylogenetic data clearly indicate that this floral trait and other morphological characters evolved in parallel. Our study illustrates that the combination of different data sets allows a comprehensive reconstruction of taxa and characteristic evolution, both of which are a precondition for future revision.

New propolis type from north-east Brazil: chemical composition, antioxidant activity and botanical origin.

BACKGROUND: Propolis is a bee product with wide diversity of biological activity. It has a complex composition, which is dependent on its botanical source. The present study aimed to determine the chemical profile, antioxidant activity and botanical origin of two samples of a propolis type from two locations of the state of Rio Grande do Norte (RN, north-east Brazil). RESULTS: The standard chemical characteristics of the RN propolis are similar or superior to the internationally marketed Brazilian green propolis. RN propolis from two locations have high antioxidant activity, corresponding to 10% (municipality of Afonso Bezerra) and 13% (municipality of Alto do Rodrigues) of quercetin activity by the 2,2-diphenyl-1-picrylhydrazyl method and to 15% (both locations) by the ß-carotene discoloration method. High-performance liquid chromatography with diode array detection (HPLC-DAD)-electrospray ionization-tandem mass spectrometry analyses revealed that most constituents of the RN propolis are flavonoids, mainly flavonols and chalcones. HPLC-DAD analysis of ethanol extracts revealed a great similarity between the chemical profile of RN propolis and shoot apices of 'jurema-preta' (Mimosa tenuiflora, Leguminosae, Mimosoideae). CONCLUSION: 'Jurema-preta' shoot apices are likely resin sources of RN propolis. The chemical characteristics and antioxidant property of RN propolis provide promising prospects for the introduction of this type of propolis into the apicultural market. © 2017 Society of Chemical Industry.

Unlocking the Karyological and Cytogenetic Diversity of Iris from Lebanon: Oncocyclus Section Shows a Distinctive Profile and Relative Stasis during Its Continental Radiation.

Despite being an important target of conservation concern and horticultural interest, Lebanese irises yet have a confusing taxonomic history and species' delimitation is often considered problematic, more especially among royal irises (Iris section Oncocyclus). Indeed, these irises of exceptionally large and spectacular flowers have radiated across Caucasus and eastern Mediterranean giving rise to a number of strict endemic taxa, many of them being considered under threat. Whilst efforts have mostly focused on clarifying the evolutionary relationships in the group based on morphological and molecular data, karyological and cytogenetic characters have been comparatively overlooked. In this study, we established for the first time the physical mapping of 35S rDNA loci and heterochromatin, and obtained karyo-morphological data for ten Lebanese Iris species belonging to four sections (Iris, Limniris, Oncocyclus and Scorpiris). Our results evidenced distinctive genomic profiles for each one of the sections, where Oncocyclus irises, while having the lowest chromosome numbers, exhibit both the highest number of 35S loci and CMA3+ sites. The continental radiation of royal irises has been accompanied by a relative karyological and cytogenetic stasis, even though some changes were observed regarding karyotype formula and asymmetry indexes. In addition to that, our results enabled taxonomic differentiation between I. germanica and I. mesopotamica-two taxa currently considered as synonyms-and highlighted the need for further studies on populations of I. persica and I. wallasiae in the Eastern Mediterranean Region.

Anatomía Floral de Peristethium leptostachyum (Loranthaceae) / Floral anatomy of Peristethium leptostachyum (Loranthaceae)

ResumenPeristethium leptostachyum es una especie hemiparásita de la familia Loranthaceae, distribuida en Colombia, Costa Rica, Ecuador, Perú, Venezuela y Panamá. Previamente tratada como Struthanthus leptostachyus, la especie fuerecientemente fue reubicada en Peristethium junto con otras que previamente estaban en los géneros Cladocolea y Struthanthus. La decisión de reconocer a Peristethium como género es controversial y fue tomada con base en caracteres de la inflorescencia y de la flor; en tanto que la monofilia de los tres géneros nombrados es incierta. En esta investigación se estudió la morfoanatomía de flores e inflorescencias de Peristethium leptostachyum, detallando la estructura del androceo y gineceo, así como los procesos de microgametogénesis y megagametogénesis; adicionalmente se realizaron comparaciones con especies afines y precisiones en relación con las diagnosis previas. Se recolectaron flores en diversas fases de desarrollo en Santa María (Boyacá-Colombia), se prepararon y analizaron bajo microscopio secciones histológicas teñidas con astrabluefucsina, además de disecciones bajo estereomicroscopio. Los resultados mostraron que P. leptostachyum comparte caracteres inflorenciales con Cladocolea (inflorescencia determinada, flor terminal ebracteada), pero también con Struthanthus (pares de tríadas a lo largo del eje, brácteas caducas y flores actinomorfas). Las flores de P. Leptostachyum de Santa María son claramente hermafroditas, con androceos y gineceos totalmente desarrollados; lo cual contradice la descripción hecha por Kuijt que reporta una condición dioica para esta especie. El androceo resultó afín al de Struthanthus vulgaris, con tapetum glandular y microsporogénesis simultánea; en contraste, Cladocolea loniceroides presenta tapetum periplasmodial y microesporogénesis sucesiva. El gineceo de P. leptostachyum, al igual que en Cladocolea, Struthanthus y Phthirusa, es unilocular con mamelón y tejido arquesporial orientado hacia el estilo, el cual es sólido y con tejido amilífero. P. leptostachyum es afín a Cladocolea loniceroides y difiere de Struthanthus vulgaris por presentar varios sacos embrionarios y pelvis (hipostasa) no lignificada. La presencia de un canal estilar sólido se propone como sinapomorfía de la tribu Psittacanthinae. Dado que P. Leptostachyum comparte caracteres anatómicos florales tanto con Cladocolea como con Strutanthus, la relación entre estos tres géneros no queda resuelta, se requieren estudios filogenéticos para establecer esta relación y poner a prueba las hipótesis de monofilia de cada uno de ellos.

AbstractPeristethium leptostachyum is a hemiparasite species of the family Loranthaceae, distributed in Colombia, Costa Rica, Ecuador, Peru, Venezuela and Panama. Previously treated as Struthanthus leptostachyus, the species was recently transferred to Peristethium together with other species of Cladocolea and Struthanthus. The present research describes the inflorescence and floral morphoanatomy of Peristethium leptostachyum, detailing the structure of the androecium and gynoecium and the processes of microgametogenesis and megagametogenesis, thus allowing comparison with Struthanthus and Cladocolea. Flowering material was collected in February and August 2012, in Santa María, Boyacá, Colombia. Histological sections were prepared and stained with astrablue-fuchsin and floral dissections were performed under a stereomicroscope. Peristethium leptostachyum shares inflorescence characters with Cladocolea (determinate inflorescence, ebracteate terminal flower), but also with Struthanthus (pairs of triads along the axis, deciduous bracts and actinomorphic flowers). The flowers of P. leptostachyum from Santa María are clearly hermaphrodites with androecium and gynoecium fully developed. This observation contradicts the description by Kuijt who reported this species to be dioecious. The androecium was observed to be similar to that of Struthanthus vulgaris, with a glandular tapetum and simultaneous microsporogenesis; in contrast, Cladocolea loniceroides has a periplasmodial tapetum and successive microsporogenesis. The gynoecium of P. leptostachyum, like that of Cladocolea, Struthanthus and Phthirusa, has a unilocular ovary with a mamelon and arquesporial tissue isoriented towards the style, which in turn is solid and amyliferous. Peristethium leptostachyum is similar to Cladocolea loniceroides and differs from Strutanthus vulgaris in presenting multiple embryo sacs and an unlignified pelvis (hipostase). The presence of a solid stylar canal is proposed as a synapomorphy of the tribe Psittacanthinae. Given that P. leptostachyum shares characters with both Cladocolea and Struthanthus generic placement cannot be clearly determined on the basis of anatomical evidence. Phylogenetic studies that include representative species of all three genera are desirable to test hypotheses of monophyly. The sexual system observed here in P. leptostachyum is different from that reported by Kuijt and more studies are needed to identify the factors (geographic, ecological, etc.) that influence this variation. Rev. Biol. Trop. 64 (1): 341-352. Epub 2016 March 01.

Fenologia e prospecção fitoquímica do jaborandi (Pilocarpus microphyllus Stapf ex Holmes) / Phenology and phytochemical prospection of Jaborandi (Pilocarpus microphyllus Stapf ex Holmes)

RESUMO O estudo fenológico tem como finalidade determinar o ritmo sazonal dos eventos do ciclo de vida da planta, como floração e frutificação. Estes eventos são determinados por uma série de fatores, como: alternância de períodos chuvosos ou não chuvosos, intensidade da radiação solar, entre outros. A fitoquímica tem por objetivos conhecer os constituintes químicos de espécies vegetais ou avaliar sua presença. O presente trabalho teve como objetivo realizar a caracterização fenológica e a prospecção fitoquímica de folhas de jaborandi. A área de estudo para a avaliação do material vegetal foi o Banco Ativo de Germoplasma de Jaborandi da Embrapa Amazônia Oriental, situada no município de Belém-PA. Os acessos escolhidos foram: Merck, cultivado a pleno sol e à sombra; Japonês e Bonal 4, cultivados a pleno sol. Os registros foram realizados diariamente por um período de 28 meses correspondendo a agosto de 2010 a dezembro de 2012, de cinco plantas/acesso e organizados para demonstração mensal, através de fichas com a numeração respectiva das plantas, com registro de presença ou ausência das fenofases, floração e frutificação. A determinação do peso seco das amostras coletadas dos acessos foi realizada no Laboratório de Agroindústria da Embrapa Amazônia Oriental, onde, após a triagem e remoção das impurezas, as folhas foram cortadas, pesadas, colocadas em bandejas de inox e secas em estufa com circulação mecânica (FANEM 320-SE), à temperatura de 45º C por 120 h. Em seguida, as amostras foram pesadas, trituradas e acondicionadas em sacos plásticos devidamente identificados e guardados sob refrigeração à temperatura de 10º C até o uso. Os extratos das plantas foram preparados utilizando-se 100 g de folhas secas de cada acesso, triturados e submetidos à extração hidroalcoólica (etanol 80%) em banho-maria sob refluxo, por aproximadamente 4 horas. Os extratos foram armazenados protegidos da luz na geladeira até o momento das análises. Foi analisada a presença das seguintes classes de substâncias químicas: ácidos orgânicos, açúcares redutores, polissacarídeos, proteínas e aminoácidos, taninos, catequinas, flavonoides, glicosídeos cardíacos, lactonas sesquiterpênicas, azulenos, carotenoides, esteroides e triterpenoides, depsídeos e depsidonas, derivados da cumarina, saponina espumídica, alcaloides, purinas, antraquinonas. Os resultados obtidos demonstraram que a espécie Pilocarpus microphyllus apresentou floração durante o ano todo e frutificação em onze meses, e a prospecção fitoquímica revelou a presença de 11 classes de constituintes químicos.

ABSTRACT Phenological studies aim to determine the seasonal rhythm of the plant life cycle events, as flowering and fruiting. These events are determined by different factors, such as: alternating periods of rainy or dryer seasons, solar radiation intensity, among others. Phytochemistry aims to identify the chemical constituents of plant species or to evaluate their presence. This study aimed the phenological characterization and the phytochemical prospection of jaborandi leaves. The chosen study area for the plant material assessment was the Active Germplasm Bank of Jaborandi in the Embrapa Eastern Amazon, located in the city of Belém, PA, Brazil. The chosen accessions were the following: Merck, grown in full sun and in shade; Japanese and Bonal 4, both grown in full sun. Records were taken on a daily basis for a period of 28 months (August of 2010 to December of 2012), from five plants/accession, and arranged for monthly demonstrations by record sheets containing the corresponding plant numeration and the presence or absence of flowering and fruiting phenophases. The dry weight of the samples collected from the accessions was measured at the Laboratory of Agribusiness of Embrapa Eastern Amazon, where, after the sorting and removal of impurities, the leaves were cut, weighed, placed in stainless steel trays, and dried in forced air circulation oven (FANEM 320 UP) at a 45°C for 120h. Then, the samples were weighed, crushed, and placed in plastic bags properly identified and stored under refrigeration at a temperature of 10ºC until the use. The plant extracts were prepared using 100g of dried leaves from each accession, crushed, and subjected to hydroalcoholic extraction (80% ethanol) with water bath heating under reflux for approximately 4 hours. The extracts were stored protected from light in a refrigerator until the analysis. We analyzed the presence of the following classes of chemical substances: organic acids, reducing sugars, polysaccharides, proteins and amino acids, tannins, catechins, flavonoids, cardiac glycosides, sesquiterpene lactones, azulenes, carotenoids, steroids and triterpernoids, depsides and depsidones, coumarin derivatives, foam saponin, alkaloids, purines, anthraquinones. Our results showed that the flowering of Pilocarpus microphyllus occurred throughout the year and fruiting occurred in eleven months, and the phytochemical prospection revealed the presence of 11 classes of chemical constituents.

Estructura floral de dos especies de Trachycarpeae (Arecaceae) / Floral structure of two species of Trachycarpeae (Arecaceae)

Copernicia and Washingtonia are two genera of the Trachycarpeae for which no subtribal classification has been proposed, mainly because of the lack of resolution in phylogenetic studies. Morphology and anatomy of flowers whithin Coryphoideae have proven useful for taxa delimitation and supporting relationships among their members. A description of the morphological and anatomical structure of flowers of C. tectorum and W. filifera is presented in order to explore reproductive characters that may clarify their classification within the subfamily and to contribute with floral biology studies. Flowers of cultivated specimens of both taxa and developing fruits of C. tectorum were fixed in FAA, dissected for morphological analysis, and parafin-embedded flowers and fruits were serially sectioned for obtaining permanent slides, using conventional techniques and safranin-fast green staining. All procedures were carried out in the Laboratory of Morpho-Anatomy, Agronomy Faculty of the Universidad Central de Venezuela (UCV). Both species have hermaphroditic flowers. C. tectorum flowers have a thick and pubescent perianth, six stamens with filaments forming a tube fused to the corolla, with rounded projections and an acute apex where the anthers are inserted. W. filifera flowers have an irregularly dentate calyx, and a shortly acuminate corolla, six stamens united by their filaments to the corolla which at the same time are briefly fused to the gynoecium. Cells with druse crystals in the staminal tube are reported for C. tectorum. Only one of the carpels of the gynoecium of C. tectorum develops at fruit stage, and a layer of abundant raphide cells forming a crustaceous endocarp in mature fruits, was found. W. filifera presents the perianth mesophyll with few layers of thick walled cells and schlerenchymatic tissue, gynoecium with apically fused carpels in the ventral region of ovary, free at the base and the apex of the style, where the ventral sutures are opened. C. tectorum has a ventral hypodermis in the petals made of large and thick walled cells, gynoecium with apically fused carpels in the ovary, free and adpressed basally, style-stigma completely fused, and stylar transmission channel absent distally. Distinct stylar canals in C. tectorum, united distally in W. filifera confirm the close relationship between these species and subtribe Livistoninae. Also, some floral morpho-anatomical similarities (e.g. fleshy calyx base and a hypodermis with thickened cell walls in petals) were found between C. tectorum and Pritchardia, supporting the affinities between both genera. Rev. Biol. Trop. 62 (3): 1137-1146. Epub 2014 September 01.

Copernicia y Washingtonia son dos de los géneros de Trachycarpeae para los cuales no ha sido propuesta una clasificación subtribal debido a la carencia de resolución en los estudios filogenéticos. La morfología y anatomía floral dentro de Coryphoideae han sido útiles para la delimitación de taxones y apoyo de relaciones. En este trabajo se presenta una descripción de la estructura morfo-anatómica de las flores de C. tectorum y W. filifera, con la finalidad de explorar caracteres reproductivos que puedan aclarar su ubicación dentro de la subfamilia y contribuir con estudios de biología floral. Se fijaron flores de especimenes cultivados de ambos taxones y frutos en desarrollo de C. tectorum, se realizaron disecciones para las descripciones morfológicas y cortes anatómicos seriados para la obtención de láminas permanentes utilizando técnicas convencionales con parafina. Todos los procedimientos fueron llevados a cabo en el Laboratorio de Morfo-Anatomía, Facultad de Agronomía de la Universidad Central de Venezuela (UCV). Ambas especies tienen flores hermafroditas. Las flores de C. tectorum tienen perianto engrosado y pubescente. Las flores de W. filifera presentan un cáliz irregularmente dentado y corola corto-acuminada. Se reportan células con drusas en el tubo estaminal de C. tectorum. Uno solo de los carpelos de C. tectorum se desarrolla en fruto y se encontró una capa de células con rafidios que podrían formar el endocarpo crustáceo en los frutos maduros. W. filifera presenta pocas capas de células de paredes engrosadas y tejido esclerenquimático en el mesofilo del perianto, gineceo con carpelos fusionados en el ápice del ovario (en su porción ventral), libres en la base y en el ápice del estilo, donde presenta suturas ventrales abiertas. Los pétalos de C. tectorum poseen una hipodermis ventral de células con paredes muy engrosadas, gineceo con carpelos fusionados en el ápice del ovario, libres y adpresos en su base, región estilar-estigmática fusionada y canal de transmisión estilar ausente distalmente. La presencia de canales estilares separados en C. tectorum, unidos distalmente en W. filifera confirman la relación cercana de estas especies con la subtribu Livistoninae. Igualmente, se encontraron algunas similitudes entre C. tectorum y Pritchardia (p.e. base carnosa del cáliz y epidermis ventral con células de paredes engrosadas en los pétalos), apoyando ciertas afinidades entre ambos géneros.

Estructura, ontogenia y vascularización de las flores e inflorescencias de Drimys granadensis (Winteraceae) / Structure, ontogeny and vascularization of the flowers and inflorescences of Drimys granadensis (Winteraceae)

Drimys granadensis is a widespread species in montane forests of South and Central America. In this research, the structure, ontogeny, phyllotaxis and vascularization of the flowers and inflorescences of this species was studied in a population from the Eastern hills of Sabana de Bogota, Colombia. The methods used applied both optical microscopy, with astra blue-fuchsin staining, and scanning electron microscopy, using critical point dryed and gold-paladium metallized samples. Besides, results were compared with those of Drimys winteri, a widely studied species distributed in Chile and Argentina. Additionally, we studied the detail of the floral anatomy to determine the bracteal or calicine identity of the caliptra. I confirmed the proliferative status of the monothelic inflorescence, discarding alternative explanations of the terminal flower identity. I found that uniflorescences have an acropetal development until the terminal meristem becomes the terminal flower, then this flower develops rapidly resulting in a determined uniflorescence. I found pseudosyphonosthelic vascularization in peduncles and pedicels. Besides, I discovered some evidence in the vascular and anatomical structures, to consider the caliptra as the fusion product of various structures and therefore of calicine origin. The caliptra showed a whorled phyllotaxis, but the petals, stamens and carpels presented a spiral condition; phyllotaxis change was explained by the long time lapse between the initiation of the calyx and the corolla. I found great similarities among the inflorescences of D. granadensis and D. winteri; they were different in the proliferation start time, and in the frequent presence of nomophylls in D. granadensis, in contrast to the presence of reduced bracts and bracteoles in D. winteri inflorescences.

Drimys granadensis es una especie de amplia distribución en los bosques montanos de Sur y Centro América. En esta investigación se estudiaron, mediante microscopía óptica y electrónica de barrido, la estructura, ontogenia, filotaxis y vascularización de sus flores e inflorescencias, y fueron comparadas con las de Drimys winteri, especie distribuida en Chile y Argentina. Adicionalmente, se buscó evidencia para determinar la identidad bracteal o calicina de la caliptra de sus flores. Se confirmó la condición monotélica proliferante de la inflorescencia, descartando explicaciones alternativas de identidad de la flor terminal. Las uniflorescencias presentan un desarrollo acrópeto, hasta que el meristemo terminal se transforma en flor terminal, entonces esta flor se desarrolla rápidamente dando lugar a una uniflorescencia determinada. La vascularización es pseudosifonostélica para pedúnculos y pedicelos. Se encontró evidencia en la vascularización y estructura anatómica para considerar la caliptra como el producto de fusión de varias estructuras y, por tanto, de origen calicino. La caliptra presentó una filotaxis verticilada, pero los pétalos, estambres y carpelos presentaron una filotaxis espiralada; el cambio se explicó por el tiempo prolongado entre la iniciación de cáliz y corola. Las inflorescencias de D. granadensis y D. winteri son muy similares; siendo diferente el tiempo de inicio de la proliferación y la frecuente presencia de nomófilos en las inflorescencias de D. granadensis, en contraste con la presencia de brácteas y bracteolas reducidas en D. winteri.

Flowers visited by hummingbirds in the open habitats of the southeastern brazilian mountaintops: species composition and seasonality / Flores visitadas por beija-flores em habitats abertos de montanhas do sudeste do Brasil: sazonalidade e composição de espécies

The hummingbird-visited plant community located on the open-habitat mountaintop of the Espinhaço Range was studied for two years (from August 2007 to July 2009) in Serra do Cipó National Park, Southeastern Brazil (19° 15′ S and 43° 31′ W). The floral characteristics and flowering period of the hummingbird-visited plants was monthly recorded along trails located in three vegetation types: (1) typical campos rupestres (TCR), (2) open fields (OPF), and (3) capões de mata (CAM). Hummingbird visitation was observed in 51 plant species, 22 ornithophilous and 29 non-ornithophilous species. The TCR showed the greatest number of species visited (N = 38), followed by the OPF (N = 18) and CAM (N = 17). Six species of hummingbirds were recorded visiting flowers: Augastes scutatus, Campylopterus largipennis, Colibri serrirostris, Chlorostilbon lucidus, Eupetomena macroura and Phaethornis pretrei. This study demonstrates that the species richness and the number of ornithophilous species visited by the hummingbirds at the study site are more similar to hummingbird-plant communities of the Atlantic Forest than to those of the Cerrado communities and other Brazilian highland open-habitat communities. The plant families most visited by hummingbirds were Bromeliaceae and Asteraceae. Although the Asteraceae family is rarely used as a food resource for hummingbirds in other high and lowland communities, in the study site this family is used mainly by the endemic hummingbird Augastes scutatus. We found a large overlap of flowering throughout the year among the species visited by the hummingbirds. Thus, the nectar availability supports these resident hummingbirds. The present study also showed that the studied hummingbird-plant community is composed of many species endemic to the campos rupestres of the Espinhaço Range, some of which are considered to be in danger of extinction, thus constituting a unique and threatened community. Thus, understanding hummingbird-plant pollination dynamics becomes fundamental to the conservation of the campos rupestres.

A comunidade de flores visitadas por beija-flores em habitats abertos de montanhas da Serra do Espinhaço, sudeste do Brasil foi estudada por dois anos (de agosto de 2007 a julho de 2009) no Parque Nacional da Serra do Cipó (19° 15′ S e 43° 31′ W). As características florais e o período de floração das plantas visitadas foram registradas mensalmente ao longo de trilhas localizadas em três tipos vegetacionais: (1) campos rupestres típicos (TCR), (2) campos abertos (OPF), e (3) capões de mata (CAM). Foram observadas visitas de beija-flores a 51 espécies de plantas: 22 ornitófilas e 29 não-ornitófilas. O TCR apresentou o maior número de espécies visitadas (N = 38), seguido pelo OPF (N = 18) e CAM (N = 17). Seis espécies de beija-flores foram observadas visitando as flores: Augastes scutatus, Campylopterus largipennis, Colibri serrirostris, Chlorostilbon lucidus, Eupetomena macroura e Phaethornis pretrei. A riqueza de espécies e o número de espécies ornitófilas visitadas pelos beija-flores, neste estudo, foi mais similar à comunidades de plantas visitadas por beija-flores na Floresta Atlântica, que as comunidades do Cerrado, bem como de outras comunidades de ambientes abertos do Brasil. As famílias com maior número de plantas visitadas pelos beija-flores foram Bromeliaceae e Asteraceae. Apesar da família Asteraceae, raramente ser utilizada como fonte alimentar pelos beija-flores em outras comunidades, nas áreas amostradas neste estudo esta família foi utilizada, principalmente, pelo beija-flor endêmico A. scutatus. Nós registramos grande sobreposição de floração ao longo do ano entre as espécies visitadas pelos beija-flores. Portanto, a disponibilidade de néctar mantem os beija-flores residentes. Este estudo, demonstrou também, que a comunidade de plantas visitadas pelos beija-flores nesta área é composta por muitas espécies endêmicas dos campos rupestres da Serra do Espinhaço, algumas das quais consideradas em perigo de extinção, constituindo assim uma comunidade única e ameaçada. Portanto, o entendimento da dinâmica de polinização desta comunidade é fundamental para a conservação dos campos rupestres.

Avaliação da atividade antibacteriana de extratos etanólico e de ciclohexano a partir das flores de camomila (Matricaria chamomilla L. ) / Evaluation of the antibacterial activity of ethanolic and cyclohexane extracts of chamomile flowers (Matricaria chamomilla L. )

Devido à crescente seleção de microrganismos resistentes aos antimicrobianos atuais, tem-se valorizado a busca por alternativas naturais. O presente estudo teve por objetivo avaliar a atividade antibacteriana de extratos etanólico e de ciclohexano de flores de camomila, espécie vegetal de uso antigo pela medicina tradicional, frente às bactérias ATCC Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli e Salmonella enterica subsp. enterica sorovar Typhimurium pelas técnicas de difusão em ágar e diluição em caldo. Foi observada inibição do crescimento de P. aeruginosa frente ao extrato etanólico bruto (1g/mL) na técnica de diluição em caldo, e confirmada pela técnica de difusão em ágar (halo de inibição de 10 mm de diâmetro). Para as demais bactérias testadas, os extratos e suas diluições não apresentaram efeito bacteriostático em nenhuma das técnicas. Pode-se concluir que o extrato etanólico bruto da camomila apresentou atividade antibacteriana frente à P. aeruginosa, porém não foi eficaz frente à S. aureus, E. coli e Salmonella enterica subsp. enterica sorovar Typhimurium. Portanto, são necessários novos estudos com diferentes linhagens de microrganismos, com o intuito de corroborar e assegurar os resultados apresentados, para definir o potencial antimicrobiano do extrato da camomila.

Due to the growing selection of microorganisms resistant to antimicrobial, the search for natural alternatives has become popular. The objective of the present study was to evaluate the antibacterial activity of ethanolic and cyclohexane extracts of chamomile flowers, a plant species long used by traditional medicine, against ATCC bacteria Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Salmonella enterica subsp. enterica serovar Typhimurium by agar diffusion and broth dilution techniques. Growth of P. aeruginosa was inhibited when crude ethanolic extract (1g/mL) was used broth dilution, and was confirmed by agar diffusion, (10 mm diameter inhibition zone). For the other bacteria tested, the extracts and their dilutions did not show any bacteriostatic effect in any of the techniques. It may be concluded that pure ethanolic extract of chamomile presents antibacterial action against P. aeruginosa, and none against S. aureus, E. coli e Salmonella enterica subsp. enterica sorovar Typhimurium. However, other studies with different strains of microorganisms may be useful in order to corroborate and ensure these results, to define evaluation of the antimicrobial activity of chamomile extract.