Interspecific insect relationships on Terminalia argentea (Myrtales: Combretaceae) trees in the Cerrado biome.

Terminalia argentea Mart. (Combretaceae), native to Brazil, is used in habitat restoration programs. Arthropods are bioindicators because their populations reflect changes in the environment. We evaluated the recovery of a degraded area by using ecological indices and analyzing arthropod interactions on T. argentea plants. The richness and diversity of sap-sucking Hemiptera and the abundance of tending ants and Sternorrhyncha predators increased with the number of T. argentea leaves. The correlation of the abundance of tending ants and Sternorrhyncha predators was positive with that of the sap-sucking Hemiptera, and the abundance of Sternorrhyncha predators was negative with that of tending ants and sap-sucking Hemiptera. The positive correlation between the abundance, richness, and diversity of insect groups and numbers of T. argentea leaves is an example of the bottom-up regulation mechanism, with the population dynamics of the lower trophic levels dictating those of higher trophic levels. The contribution of T. argentea, a host plant of many arthropods, to the recovery of ecological relationships between organisms in degraded ecosystems is important.

Insecticidal efficacy and possibility of Combretum trifoliatum Vent. (Myrtales: Combretaceae) extracts in controlling Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae).

BACKGROUND: The fall armyworm Spodoptera frugiperda (J.E. Smith), is an important pest of agronomical crops. It is interesting to discover secondary metabolites in plants that are environmentally safer than synthetic pesticides. For this purpose, Combretum trifoliatum crude extract and its isolated compounds were investigated for their insecticidal activities against S. frugiperda. RESULTS: The median lethal dose (LD50 ) was evaluated in the second-instar larvae using the topical application method. The isolated compounds, apigenin and camphor, demonstrated a highly toxic effect on larvae at a lower LD50 dose than crude extract. Moreover, when the larvae were exposed to crude extract concentrations, the development to pupa and adult stages was reduced by more than 50%. The ovicidal toxicity was examined using a hand sprayer. The extract concentration 5, 10, and 20 µg/egg significantly decreased the egg hatchability. In addition, crude extract showed a significant difference in inhibiting acetylcholinesterase (AChE) activity while crude extract and camphor showed significant inhibitory effects on carboxylesterase (CE) and glutathione-S-transferase (GST) activities. CONCLUSION: The crude ethanol extract of Combretum trifoliatum was toxic to S. frugiperda in terms of larval mortality, negatively affecting biological parameters, and decreasing egg hatchability. Additionally, the activities of cholinergic and detoxifying enzymes were affected by crude extract and its isolated compounds. These results highlight that Combretum trifoliatum might be efficient as a bioinsecticide to control S. frugiperda. © 2023 Society of Chemical Industry.

Absence of aluminium compromises root integrity, reduces leaf hydration and Rubisco performance in Qualea grandiflora, an Al-accumulating species.

Aluminium (Al) is toxic to most plants. Nevertheless, some species accumulate Al without showing toxicity symptoms. Previous studies have evidenced Al in chloroplasts of Al-accumulating species from the Cerrado vegetation in South America. We ask whether Al increases carbon assimilation through enhanced apparent efficiency of Rubisco. Seedlings of the Al-accumulator Qualea grandiflora (Vochysiaceae) were grown in nutrient solution with 0, 740, and 1480 µm Al. Growth parameters, relative leaf water content, Al concentration in organs, gas exchange and apparent carboxylation efficiency (measured from A/Ci curves) were evaluated for 60 days. Plants without Al showed no root growth, necrotic roots, low gas exchange rates, and decreased apparent carboxylation efficiency. Al-treated plants, however, showed new white roots and increased root biomass leading to higher leaf hydration, and apparent carboxylation efficiency was higher in these plants. Increased Al available in the nutrient solution increased Al accumulation in plant organs. Absence of Al compromised root integrity in Q. grandiflora, thus limiting leaf hydration. No positive direct effect of Al on Rubisco was evidenced in Al-treated plants.

Feno-morfología de dos especies de páramo de Miconia (Melastomataceae): optimización de la floración y fructificación para mejorar la dispersión / Pheno-morphology of two paramo species of Miconia (Melastomataceae): optimization of flowering and fructification to enhance dispersal

Introducción: El Páramo es uno de los ecosistemas más afectados por actividades humanas, lo que aumenta la necesidad de estudios fenológicos como base para el manejo y la conservación. Objetivo: Describir la feno-morfología de Miconia ligustrina y Miconia elaeoides. Métodos: De septiembre 2019 a febrero 2020, y de abril 2021 hasta agosto del mismo año, se monitoreó la fenología de 12 individuos de cada especie. Adicionalmente, se vincularon datos fenológicos de ejemplares de herbario, para corroborar los resultados obtenidos. En los dos conjuntos de datos, se describió su distribución usando estadística circular, además, se obtuvieron registros de precipitación y temperatura del área. Adicionalmente, se describió la morfometría de cada especie. Resultados: El pico de floración de M. lingustrina es en abril, y la fructificación se concentra entre junio y julio; M. elaoides, florece de forma masiva en enero, con la máxima producción de frutos en mayo. Hubo correlación entre fenología y lluvias: la floración se da en época seca y la fructificación en la época lluviosa. Las especies difieren en morfología, pero, como en otras especies de Miconia, las flores y frutos son pequeños (aprox. 5 mm). Conclusión: La floración se presenta en la época seca y la fructificación en la época lluviosa, cuando las tasas de fotosíntesis son bajas y la frugivoría incrementa. Este patrón fenológico favorece la dispersión óptima de semillas.

Introduction: Paramos are among the ecosystems that are most affected by anthropic activities, increasing the need for phenological studies as a basis for management and conservation. Objective: To describe the pheno-morphology of Miconia ligustrina and Miconia elaeoides. Methods: From September 2019 to February 2020, and from April 2021 to August of the same year, the phenology of 12 individuals of each species was monitored. Additionally, phenological data of herbariums were linked to corroborate obtained results. In the two data sets, distribution of data was described using circular statistics; in addition, we recorded the precipitation and temperature in the area. Additionally, we described the morphometry of each species. Results: The flowering peak of M. ligustrina is in April, and fruiting concentrates between June and July; in M. elaeoides, flowering is massive in January, and the highest fruit production in May. The phenology was correlated with rainfall: flowering takes place in the dry season and fruiting in the rainy season. The species differ morphologically, but, as in other Miconia species, the flowers and fruits are small (approx. 5 mm). Conclusions: Flowering occurs in the dry season and fructification in the rainy season, when the photosynthetic rate is low and frugivory increases. This phenological pattern favors optimal seed dispersal.

Metabolites in Conocarpus erectus leaves attenuate α-amylase activity by modulating amino acid residues of α-amylase: an in vitro and docking study / Los metabolitos en las hojas de Conocarpus erectus atenúan la actividad de la α-amilasa al modular los residuos de aminoácidos de la α-amilasa: un estudio in vitro y de acoplamiento

The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 ́-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.

Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 ́-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.

Leaf extract of Osbeckia octandra induces apoptosis in oral squamous cell carcinoma cells.

BACKGROUND: Osbeckia octandra is a plant endemic to Sri Lanka and is used in ethnomedicine for treating various diseases. However, the anti-cancer properties of O. octandra are yet to be fully investigated. In the present study, we evaluated the anti-cancer effects of O. octandra on oral cancer cells. METHODS: Human oral cancer cell lines (HSC2, YD10B, YD38, YD9, and YD32) were used in this study. BrdU incorporation, cell cycle and annexin-V/PI staining were all evaluated using flow cytometry to determine the extent to which O. octandra leaf extract inhibits cell proliferation and induces apoptosis. Cell viability and reactive oxygen species (ROS) were also measured in order to investigate the anti-cancer effects of O. octandra extracts. Western blotting was performed to detect cell cycle related protein such as cyclin d1 and cdk4, and to detect apoptosis-related proteins such as Bcl-2, Bcl-XL, Bax, Caspase-9, Cleaved caspase-3, Fas, Caspase-8, and Bid. RESULTS: Leaf extract of O. octandra reduced oral squamous cell carcinoma (OSCC) cell viability in a dose-dependent manner. Leaf extract of O. octandra has non-toxic in normal keratinocytes. Also, O. octandra extract interrupted the DNA replication via G1 phase arrests, and this effect was independent of ROS generation. In the apoptosis-related experiments, the population of annexin V-positive cells increased upon treatment with O. octandra extract. Furthermore, the expression of anti-apoptotic protein (Bcl-2 and Bcl-xL) was decreased, whereas the expression of cleaved caspase-3 protein was increased in O. octandra-treated OSCC cells. CONCLUSIONS: The results suggest that a leaf extract of O. octandra inhibited the proliferation of OSCC cells through G1 phase arrest and interrupting DNA replication. The leaf extract of O. octandra could trigger the apoptotic response via caspase 3 activation in OSCC cells. These results suggest that O. octandra has the potential to be developed as an alternative medicine for treating OSCC.

TLC-Based Fingerprinting Analysis of the Geographical Variation of Melastoma malabathricum in Inland and Archipelago Regions: A Rapid and Easy-to-Use Tool for Field Metabolomics Studies.

Melastoma malabathricum is an Indo-Pacific herb that has been used traditionally to treat numerous ailments such as wounds, dysentery, diarrhea, toothache, and diabetes. The objective of this study was to evaluate the variability of the metabolic profiles of M. malabathricum across its geographic distribution. By employing thin layer chromatography (TLC), specimens collected from six terrestrial and archipelago regions of Indonesia were analyzed by densitometry for metabolomic fingerprinting analysis combined with chemometric tools: principal component analysis (PCA) and hierarchical cluster analysis (HCA). Two PCAs were identified as PC1 and PC2 with 41.90% and 20.36%, respectively. Our results indicate the importance of considering geographic distribution during field-collection efforts since they demonstrate regional metabolic variation in secondary metabolites of M. malabathricum, as illustrated by TLC and their biological activities.

The Melastoma dodecandrum genome and the evolution of Myrtales.

Melastomataceae has abundant morphological diversity with high economic and ornamental merit in Myrtales. The phylogenetic position of Myrtales is still contested. Here, we report the chromosome-level genome assembly of Melastoma dodecandrum in Melastomataceae. The assembled genome size is 299.81 Mb with a contig N50 value of 3.00 Mb. Genome evolution analysis indicated that M. dodecandrum, Eucalyptus grandis, and Punica granatum were clustered into a clade of Myrtales and formed a sister group with the ancestor of fabids and malvids. We found that M. dodecandrum experienced four whole-genome polyploidization events: the ancient event was shared with most eudicots, one event was shared with Myrtales, and the other two events were unique to M. dodecandrum. Moreover, we identified MADS-box genes and found that the AP1-like genes expanded, and AP3-like genes might have undergone subfunctionalization. The SUAR63-like genes and AG-like genes showed different expression patterns in stamens, which may be associated with heteranthery. In addition, we found that LAZY1-like genes were involved in the negative regulation of stem branching development, which may be related to its creeping features. Our study sheds new light on the evolution of Melastomataceae and Myrtales, which provides a comprehensive genetic resource for future research.

A new species of gall midge (Diptera, Cecidomyiidae) associated with Pleroma raddianum (DC) Gardner (Myrtales: Melastomatacea), an endenic plant to Brazil

Lopesia pleromatis sp. nov. (Lopesiini, Cecidomyiidi) is described based on material collected in Atlantic Forest areas of Bertioga (São Paulo State, Brazil). Specimens were obtained from globoid leaf galls on Pleroma raddianum (DC.) Gardner (Melastomataceae), an endemic plant to Brazil. Lopesia pleromatis is compared to other congeneric species. The most important morphological characters are illustrated.(AU)

A nuclear phylogenomic study of the angiosperm order Myrtales, exploring the potential and limitations of the universal Angiosperms353 probe set.

PREMISE: To further advance the understanding of the species-rich, economically and ecologically important angiosperm order Myrtales in the rosid clade, comprising nine families, approximately 400 genera and almost 14,000 species occurring on all continents (except Antarctica), we tested the Angiosperms353 probe kit. METHODS: We combined high-throughput sequencing and target enrichment with the Angiosperms353 probe kit to evaluate a sample of 485 species across 305 genera (76% of all genera in the order). RESULTS: Results provide the most comprehensive phylogenetic hypothesis for the order to date. Relationships at all ranks, such as the relationship of the early-diverging families, often reflect previous studies, but gene conflict is evident, and relationships previously found to be uncertain often remain so. Technical considerations for processing HTS data are also discussed. CONCLUSIONS: High-throughput sequencing and the Angiosperms353 probe kit are powerful tools for phylogenomic analysis, but better understanding of the genetic data available is required to identify genes and gene trees that account for likely incomplete lineage sorting and/or hybridization events.

Comparative analysis of chloroplast genome structure and molecular dating in Myrtales.

BACKGROUND: Myrtales is a species rich branch of Rosidae, with many species having important economic, medicinal, and ornamental value. At present, although there are reports on the chloroplast structure of Myrtales, a comprehensive analysis of the chloroplast structure of Myrtales is lacking. Phylogenetic and divergence time estimates of Myrtales are mostly constructed by using chloroplast gene fragments, and the support for relationships is low. A more reliable method to reconstruct the species divergence time and phylogenetic relationships is by using whole chloroplast genomes. In this study, we comprehensively analyzed the structural characteristics of Myrtales chloroplasts, compared variation hotspots, and reconstructed the species differentiation time of Myrtales with four fossils and one secondary calibration point. RESULTS: A total of 92 chloroplast sequences of Myrtales, representing six families, 16 subfamilies and 78 genera, were obtained including nine newly sequenced chloroplasts by whole genome sequencing. Structural analyses showed that the chloroplasts range in size between 152,214-171,315 bp and exhibit a typical four part structure. The IR region is between 23,901-36,747 bp, with the large single copy region spanning 83,691-91,249 bp and the small single copy region spanning 11,150-19,703 bp. In total, 123-133 genes are present in the chloroplasts including 77-81 protein coding genes, four rRNA genes and 30-31 tRNA genes. The GC content was 36.9-38.9%, with the average GC content being 37%. The GC content in the LSC, SSC and IR regions was 34.7-37.3%, 30.6-36.8% and 39.7-43.5%, respectively. By analyzing nucleotide polymorphism of the chloroplast, we propose 21 hypervariable regions as potential DNA barcode regions for Myrtales. Phylogenetic analyses showed that Myrtales and its corresponding families are monophyletic, with Combretaceae and the clade of Onagraceae + Lythraceae (BS = 100%, PP = 1) being sister groups. The results of molecular dating showed that the crown of Myrtales was most likely to be 104.90 Ma (95% HPD = 87.88-114.18 Ma), and differentiated from the Geraniales around 111.59 Ma (95% HPD = 95.50-118.62 Ma). CONCLUSIONS: The chloroplast genome structure of Myrtales is similar to other angiosperms and has a typical four part structure. Due to the expansion and contraction of the IR region, the chloroplast genome sizes in this group are slightly different. The variation of noncoding regions of the chloroplast genome is larger than those of coding regions. Phylogenetic analysis showed that Combretaceae and Onagraceae + Lythraceae were well supported as sister groups. Molecular dating indicates that the Myrtales crown most likely originated during the Albian age of the Lower Cretaceous. These chloroplast genomes contribute to the study of genetic diversity and species evolution of Myrtales, while providing useful information for taxonomic and phylogenetic studies of Myrtales.

Advanced characterization of polyphenols from Myrciaria jaboticaba peel and lipid protection in in vitro gastrointestinal digestion.

Ultrasound-assisted and solvent extractions resulted in similar levels of hydrolyzable tannins (10.3-6.0 mg/g), anthocyanins (7.8-10.2 mg/g) and flavonols (0.24-0.32 mg/g) for dried Myrciaria jaboticaba peel (DJP). Ultrasound was efficient for the extraction of poorly soluble hydrolyzable tannins but affected the stability of anthocyanins and flavonols. UPLC-DAD-MSn allowed the identification of 44 hydrolyzable tannins as single and mixed hexosides bearing galloyl, HHDP and tergalloyl units. Twelve mixed HHDP-galloylgluconic acids and tergalloylated hexosides were newly discovered in this work. Acid hydrolysis of both ultrasonic extract and DJP yielded five major compounds, i.e. gallic acid, ellagic acid, gallic acid-C-hexoside, valoneic acid dilactone and sanguisorbic acid dilactone and pointed to higher contents in hydrolyzable tannins than by summing individual polyphenols after UPLC. Last, cyanidin-3-O-glucoside and hydrolyzable tannins from the ultrasonic extract inhibited lipid peroxidation of a Western type meal in in vitro digestion, suggesting a health benefit for these jabuticaba polyphenols.

Vochysia tucanorum Mart. butanol fraction presents antitumoral activity in vivo and prevents the installation of cachexia in solid Ehrlich tumor model.

BACKGROUND: Cancer is a multifactorial disease caused by uncontrolled proliferation of cells. About 50-80% of cancer patients develop cachexia, a complex metabolic syndrome associated with an increase of mortality and morbidity. However, there are no effective therapies in medical clinic for cancer cachexia. Vochysia tucanorum Mart. is a common three of the Brazilian "Cerrado". The butanolic fraction of V. tucanorum (Fr-BuVt), very rich in triterpenes with various biological activities, might be interesting in being tested in cancer cachexia syndrome. Hence, the present study was undertaken to investigate the antitumoral activity of Fr-BuVt and its potential against cachexia development. METHODS: Ehrlich tumor was used as model of cancer cachexia. Ascitic Ehrlich tumor cells were collected, processed and inoculated subcutaneously in saline solution (1 × 107/100 µl; ≥95% viability) for the obtention of solid Ehrlich carcinoma. After inoculation, solid Ehrlich carcinoma-bearing mice were treated by 14 consecutive days by gavage with Fr-BuVt (200 mg/kg). Body weight and tumor volume were measure during the treatment period. Tumors were removed, weighed and properly processed to measure the content and phosphorylation levels of key-proteins involved to apoptotic and proliferation process by Western Blot. Muscles and adipose tissues were removed for weighed. Serum was collected to cytokines levels and energetic blood markers measurements. RESULTS: The treatment with the Fr-BuVt (200 mg/kg, 14 days) decreased the solid Ehrlich tumor volume and weight besides increased the expression of the pro-apoptotic proteins caspase-3 and BAX, but also decreased the expression of the proteins involved in proliferation NFκB, mTOR and ERK. In addition, our data shows that the administration of Fr-BuVt was able to prevent the installation of cancer cachexia in Ehrlich carcinoma-bearing mice, since prevented the loss of body weight, as well as the loss of muscle and adipose tissue. Moreover, an improvement in some blood parameters such as decrease in cytokines TNF-α and IL-6 levels is observed. CONCLUSIONS: The study revealed that Fr-BuVt has antitumoral activity and prevent installation of cancer cachexia in Ehrlich model. Therefore, Fr-BuVt may represent an alternative treatment for cancer cachexia.

Phytochemical Screening, Cytotoxicity and Anti-inflammatory Activities of the Leaf Extracts from Lawsonia inermis of Indian Origin to Explore their Potential for Medicinal Uses.

BACKGROUND: Lawsonia inermis Linn popularly known as Henna, plays an important role in ayurvedic or natural herbal medicines. The presence of phytoconstituents in henna, that may affect the animal or human health adversely, needs to be elucidated for L. inermis Linn species grown in India. INTRODUCTION: The aim of this research was to perform phytochemical screening, and study cytotoxicity and anti-inflammatory activities to understand the potential of leaves of Lawsonia inermis of Indian origin to provide a way forward for therapeutic use in medicine. METHODS: We assessed the phytochemical profile for the presence of phytoconstituents (alkaloids, carbohydrates, glycosides, steroids, flavonoids, saponins, tannins, proteins/amino acids and gums/mucilage) in various extracts of the plant leaves. The extracts were further purified by column chromatography for the isolation of plant constituents and monitored by TLC, analyzed by Fourier transform infrared FT-IR spectroscopy, H1NMR, and GC-MS analysis. Fractions were assessed for cytotoxicity and anti-inflammatory properties at various concentrations. We assessed the anti-inflammatory activity by nitric oxide production in various leaf extracts determined by Griess assay. RESULTS: All the spectral results suggest that the compounds from the extract contain an aromatic nucleus and OH group along with the methoxy group, allyl as well as vinyl group. Fractions of chloroform/methanolic (7:3) leaf extract of Lawsonia inermis confirmed the presence of the two constituents i.e. fraxetin and 1(3H)-isobenzofuranone. We observed a significant difference in cytotoxicity at higher concentrations in methanol and chloroform: methanol (8:2) leaf extracts (p>0.05), we could not find any significant differences amongst other leaf extracts at different concentrations. Some leaf extracts have potential cytotoxic activity on Vero cells. Reducing the chloroform concentration during extraction decreases the cytotoxic effect on cells. Nitric oxide levels decreased from 1000 µg/ml concentration to lower concentrations with varying degrees. Overall the highest nitric oxide production by CHCl3 (70%)/ MeOH (30%) was observed amongst various fractions at different concentrations. CONCLUSION: Phytochemical screening and the study of cytotoxicity and anti-inflammatory activities highlight the potential of leaves of the plant to provide a way further for their use in medicine. Fraxetin 1(3H) and isobenzofuranone structures were confirmed in fractions of CHCl3 (70%)/ MeOH (30%) extract as potent constituents. Some leaf extracts have potential cytotoxic activity on Vero cells. Reducing the chloroform concentration during extraction, it decreases the cytotoxic effect on cells. The cytotoxicity studies indicate the presence of cytotoxic compounds in some of these extracts, warranting research for the fabrication of suitable formulations comprising these constituents to reduce dose/toxicity for beneficial effects of the plant components.

Ambrosia Beetle (Coleoptera: Curculionidae) Communities and Frass Production in 'Ohi'a (Myrtales: Myrtaceae) Infected With Ceratocystis (Microascales: Ceratocystidaceae) Fungi Responsible for Rapid 'Ohi'a Death.

Rapid 'Ohi'a Death (ROD) is a deadly disease that is threatening the native Hawaiian keystone tree species, 'ohi'a lehua (Metrosideros polymorpha Gaudich). Ambrosia beetles (Curculionidae: Scolytinae) and their frass are hypothesized to play a major role in the spread of ROD, although their ecological niches and frass production within trees and across the landscape are not well understood. We characterized the beetle communities and associated frass production from bolts (tree stem sections) representative of entire individual 'ohi'a trees from multiple locations across Hawai'i Island by rearing beetles and testing their frass for viable ROD-causing fungi. Additionally, we estimated frass production for three beetle species by weighing their frass over time. We found that Xyleborinus saxesenii (Ratzburg), Xyleborus affinis Eichhoff, Xyleborus ferrugineus (Fabricius), Xyleborus perforans (Wollaston), and Xyleborus simillimus Perkins were commonly found on ROD-infected 'ohi'a and each produced frass containing viable Ceratocystis propagules. The Hawai'i Island endemic beetle and the only native ambrosia beetle associated with 'ohi'a, X. simillimus, was limited to high elevations and appeared to utilize similar tree heights or niche dimensions as the invasive X. ferrugineus. Viable Ceratocystis propagules expelled in frass were found throughout entire tree bole sections as high as 13 m. Additionally, we found that X. ferrugineus produced over 4× more frass than X. simillimus. Our results indicate the ambrosia beetle community and their frass play an important role in the ROD pathosystem. This information may help with the development and implementation of management strategies to control the spread of the disease.

Historical biogeography of Vochysiaceae reveals an unexpected perspective of plant evolution in the Neotropics.

PREMISE: Despite the fast pace of exploration of the patterns and processes influencing Neotropical plant hyperdiversity, the taxa explored are mostly from large groups that are widely distributed, morphologically diverse, or economically important. Vochysiaceae is an example of an undersampled taxon, providing an excellent system for investigating Neotropical biogeography. We present a phylogenomics-based hypothesis of species relationships in Vochysiaceae to investigate its evolutionary history through space and time. METHODS: We inferred a phylogeny for 122 species from Vochysiaceae and seven other families of Myrtales. Fossils from four myrtalean families were used to estimate the divergence times within Vochysiaceae. Historical biogeography was estimated using ancestral range probabilities and stochastic mapping. RESULTS: Monophyly of all genera was supported except for Qualea, which was split by Ruizterania into two clades. Vochysiaceae originated ~100 mya, splitting into an Afrotropical and a Neotropical lineage ~50 mya, and its ancestral range is in the area currently occupied by the Cerrado. CONCLUSIONS: The most recent common ancestor of Vochysiaceae + Myrtaceae had a West Gondwanan distribution, supporting a South American + African ancestral range of Vochysiaceae. On a global scale, geographic range reduction was the principal biogeographic event. At a finer scale, initial range reduction was also important and the Cerrado region was the most ancestral area with multiple colonization events to the Amazon, Central America, and the Atlantic Forest. Colonization events occurred from open areas to forest vegetation, an unusual finding regarding the evolution of plants in the Neotropics.

Adulticidal Activity of Melaleuca alternifolia (Myrtales: Myrtaceae) Essential Oil With High 1,8-Cineole Content Against Stable Flies (Diptera: Muscidae).

The stable fly, Stomoxys calcitrans (Linnaeus 1758), is a hematophagous fly responsible for causing loss of performance in horses, causing losses in cattle productivity, and impacting the animals' health through the spread of pathogenic microorganisms. The objective of this work was to investigate the insecticidal activity of essential oil obtained from Melaleuca alternifolia (Cheel), presenting high 1,8-cineole content, against S. calcitrans adults. Insecticidal activity was determined using surface application methods and exposure to oil impregnated paper. It was observed that treatments at 25 and 50 µg/cm2 (P < 0.05) present fumigant activity through exposure to the impregnated paper, and in the first 15 min of exposure, the mortality rates obtained for these treatments were, respectively (96.6 ± 3.3% and 100%), equivalent to the positive control. Using the superficial application method, the only treatment concentration presenting adulticidal action was 5% (w/v) (P < 0.05). Respective toxicities LC50 (%, w/v) and LC80 for the impregnated paper method were 1.06 ± 0.02 and 1.47 ± 0.17; for the superficial application method, they were 3.82 ± 0.65 and 5.53 ± 0.74. As demonstrated, M. alternifolia essential oil presents adulticidal potential against S. calcitrans.

Immunostimulatory and antioxidant activities of a lignin isolated from Conocarpus erectus leaves.

In this work, we investigated the antioxidant and immunomodulatory activities of a lignin isolated from Conocarpus erectus leaves. The lignin was characterized by FTIR, 1H NMR, 13C NMR and gel permeation chromatography analysis as well as ultraviolet/visible absorption spectra. The lignin was evaluated for total antioxidant activity (TAA), DPPH and ABTS+ scavenging abilities, and by a lipid peroxidation inhibition assay. Immunomodulatory activity of the lignin (10 µg/mL) on human peripheral blood mononuclear cells (PBMCs) was determined. The C. erectus lignin was found to be of the guaiacyl-syringyl-p-hydroxyphenyl (G-S-H) type, with an average molecular weight of 2709 Da (polydispersity index: 2.1). It showed low TAA (17.92%) and moderate antioxidant activity against DPPH and ABTS+ (IC50: 231.16 and 356.03 µg/mL, respectively). It also inhibited lipid peroxidation by 42.14%. The lignin promoted an increase in mitochondrial ROS levels as well as cytosolic Ca2+ in PBMCs. In addition, it promoted the differentiation and activation of CD8+ T lymphocytes, differentiation of CD14+ monocytes, and stimulated the release of nitric oxide and cytokines, mainly those linked to a Th1 response. The results showed that the C. erectus lignin may be used in future studies in which the modulation of the immune response is a key factor.

Reproductive biology and pollinators in two enantiostylous Qualea species (Vochysiaceae) in the Brazilian Cerrado.

Enantiostyly is a floral polymorphism in which two floral forms in the same species differ in deflection of the stigma to right or left position. In monomorphic enantiostylous plants, flowers of the two morphs occur within the same individual, usually in the same proportion. In self-compatible species the function of monomorphic enantiostyly is proposed to increase outcrossing rates and offer a reproductive advantage under pollination limitation. Enantiostylous species are usually self-compatible and show heteranthery, with poricide anthers and pollen as pollinator reward; however, there are families, such as Vochysiaceae, that have different characteristics. We analysed the reproductive system and pollination biology of Qualea parviflora and Q. multiflora, two enantiostylous species from the Brazilian Cerrado that have specific morphological and physiological traits. For this, we characterized flower traits, performed hand pollinations and studied floral visitors. We found no differences between morphs in the proportion of flowers, nectar produced or its concentration, pollen quantity and fruit set. Both species were self-incompatible and quite generalist regarding floral visitors. Enantiostyly in self-incompatible plants seems to confer a reproductive advantage by reducing self-interference resulting from stigma clogging. This novel result helps to expand our knowledge on this complex floral polymorphism and opens new avenues for future research on this topic.

Removal of lead, zinc and cadmium from contaminated soils with two plant extracts: Mechanism and potential risks.

Screening appropriate washing agents to remediate soils contaminated with heavy metals is crucial for decreasing metal hazards posing to environment and human health. In this study, two plant washing agents-water-extracted from Fagopyrum esculentum and Fordiophyton faberi, were applied to remove soil Pb, Zn, and Cd by washing. Results indicated that metal removals augmented with increase of washing solution concentrations, decreased with increasing pH values of the solution and followed the pseudo-second-order model depending on contact duration. At concentration of 50 g/L, pH 3 and contact duration of 120 min, F. esculentum had higher removals of Pb (5.98-6.83%), Zn (21.82-27.94%), and Cd (39.90-40.74%) than those of F. faberi. And metal ions could be removed by binding with carboxyl, hydroxyl, amide, amine and aromatic groups in washing solutions. The potential risks of residual metals declined by 51.35-52.12% for mine soil and 48.51-49.96% for farmland soil with exchangeable and carbonate-bound fractions obviously extracted after a single washing (P < 0.05). And soil organic carbon and nutrients increased to some extent except for total phosphorus and available potassium. Moreover, soil phytotoxicity lowered except that some adverse effects on seed germination existed. Therefore, the water extract from F. esculentum is a promising washing agent for heavy metal removal.