Anti-Zika Virus Activity of Plant Extracts Containing Polyphenols and Triterpenes on Vero CCL-81 and Human Neuroblastoma SH-SY5Y Cells.

Zika virus (ZIKV) infection is a global threat associated to neurological disorders in adults and microcephaly in children born to infected mothers. No vaccine or drug is available against ZIKV. We herein report the anti-ZIKV activity of 36 plant extracts containing polyphenols and/or triterpenes. ZIKV-infected Vero CCL-81 cells were treated with samples at non-cytotoxic concentrations, determined by MTT and LDH assays. One third of the extracts elicited concentration-dependent anti-ZIKV effect, with viral loads reduction from 0.4 to 3.8 log units. The 12 active extracts were tested on ZIKV-infected SH-SY5Y cells and significant reductions of viral loads (in log units) were induced by Maytenus ilicifolia (4.5 log), Terminalia phaeocarpa (3.7 log), Maytenus rigida (1.7 log) and Echinodorus grandiflorus (1.7 log) extracts. Median cytotoxic concentration (CC50 ) of these extracts in Vero cells were higher than in SH-SY5Y lineage. M. ilicifolia (IC50 =16.8±10.3 µg/mL, SI=3.4) and T. phaeocarpa (IC50 =22.0±6.8 µg/mL, SI=4.8) were the most active extracts. UPLC-ESI-MS/MS analysis of M. ilicifolia extract led to the identification of 7 triterpenes, of which lupeol and a mixture of friedelin/friedelinol showed no activity against ZIKV. The composition of T. phaeocarpa extract comprises phenolic acids, ellagitannins and flavonoids, as recently reported by us. In conclusion, the anti-ZIKV activity of 12 plant extracts is here described for the first time and polyphenols and triterpenes were identified as the probable bioactive constituents of T. phaeocarpa and M. ilicifolia, respectively.

Endophytic fungus diversity in soybean plants submitted to conditions of elevated atmospheric CO2 and temperature.

Global climatic changes can have drastic impacts on plant species, including severe consequences for the agricultural species productivity. Many of these species present important mutualisms with endophytic fungi that positively influence their performance. The present study evaluated whether the increases in CO2 and temperature predicted for the year 2100 may cause changes in foliar carbon (C) and nitrogen (N) concentrations in soybean (Glycine max) and, consequently, the interactions with its endophytic fungi. The effects of elevated CO2 and temperature were evaluated in four treatments in open-top chambers: (i) control, (ii) increased temperature, (iii) increased CO2, and (iv) increased CO2 and temperature. Increased atmospheric CO2 resulted in decreased foliar N concentration, while increased temperature increased it. A total of 16 taxa of endophytic fungi were identified based on sequencing internal transcribed spacer regions of rRNA subunits. Increased atmospheric CO2 and temperature were observed to potentially modify the endophytic mycobiota of soybean plants. The results suggest that the fungi species substitution is a consequence of changes in foliar N concentration and C/N ratio. Predicted climatic changes shall affect the relationships between plant and endophytes, which in turn, will affect the performance and resistance of soybean, one of the most important crops in the world.

The Program for Biodiversity Research in Brazil: The role of regional networks for biodiversity knowledge, dissemination, and conservation.

The Program for Biodiversity Research (PPBio) is an innovative program designed to integrate all biodiversity research stakeholders. Operating since 2004, it has installed long-term ecological research sites throughout Brazil and its logic has been applied in some other southern-hemisphere countries. The program supports all aspects of research necessary to understand biodiversity and the processes that affect it. There are presently 161 sampling sites (see some of them at Supplementary Appendix), most of which use a standardized methodology that allows comparisons across biomes and through time. To date, there are about 1200 publications associated with PPBio that cover topics ranging from natural history to genetics and species distributions. Most of the field data and metadata are available through PPBio web sites or DataONE. Metadata is available for researchers that intend to explore the different faces of Brazilian biodiversity spatio-temporal variation, as well as for managers intending to improve conservation strategies. The Program also fostered, directly and indirectly, local technical capacity building, and supported the training of hundreds of undergraduate and graduate students. The main challenge is maintaining the long-term funding necessary to understand biodiversity patterns and processes under pressure from global environmental changes.

Dissimilar forests along the Rio Doce watershed call for multiple restoration references to avoid biotic homogenization.

An environmental disaster caused by the rupture of a mining tailings dam has impacted a large area of the Rio Doce watershed in the Brazilian Atlantic Forest, resulting in unprecedented damage at spatial and temporal scales. The Atlantic Forest is one of the world's most important biodiversity hotspots. A long history of land use conversion has resulted in a highly fragmented landscape. Despite numerous restoration initiatives, these efforts have often biased criteria and use limited species assemblages. We conducted a comprehensive synthesis of the plant community in riparian forests along the Rio Doce watershed. Our work detailed vegetation composition (tree and sapling strata) and examined its relationship with edaphic and landscape factors, aiming to inform restoration projects with scientifically robust knowledge. A total of 4906 individuals from the tree strata and 4565 individuals from the sapling strata were recorded, representing a total of 1192 species from 75 families. Only 0.8% of the tree species and 0.5% of the sapling species occurred in all sampled sectors, with over 84% of the species occurring in a single watershed sector for both strata. We observed a high species heterogeneity modulated by turnover (92.3% in the tree, and 92.7% in the sapling strata) among sites. Overall, our research revealed a gradient of soil fertility influencing species composition across different strata. Additionally, we discovered that preserved landscapes had a positive impact on species diversity within both strata. The species exclusivity in the sampled sites and the high turnover rate imply the need to consider multiple reference ecosystems when restoring the watershed to reduce the risk of biotic homogenization. Finally, the reference ecosystems defined here serve as a basis for the selection of locally particular species in the implementation of restoration projects that aim to improve biodiversity, ecosystem services, and water security.

Patterns in Wing Morphology and Fluctuating Asymmetry in Eulaema nigrita along an Altitudinal Gradient in the Brazilian Rupestrian Grassland.

Mountain ecosystems experience abrupt abiotic changes that represent environmental filters for many organisms, shaping their phenotypic expressions. However, little is known about the morphological and symmetric adjustments of native bees along altitudinal gradients. We evaluated the changes on wing morphology, wing size, and vein fluctuating asymmetry (FA) of Eulaema nigrita Lepeletier (Apidae: Euglossini) associated with climatic variables along an altitudinal gradient in the rupestrian grassland (known also as campo rupestre or rupestrian field) of Serra do Cipó, Brazil. Seven sampling points along the altitudinal gradient were selected and distributed among 800 and 1400 m.a.s.l., and then, 40 individuals of E. nigrita were collected per each altitudinal point to determine the FA levels and the morphological changes using geometric morphometric techniques. We found that the wing size of E. nigrita decreased with increasing altitude. At the highest altitudes, the levels of FA of the wing veins were greater compared to bees from lower altitudes. We detected significant changes in wing morphology along the altitudinal gradient; bees of lower altitudes showed longer and wider wings than bees of higher altitudes, which had narrower and less elongated wings. Our results show a set of morphological adjustments and phenotypic expressions in E. nigrita associated with the variation in environmental conditions along the altitudinal gradient. We highlight the importance of environmental variables as insect-stressor factors, and that FA and geometric morphometric can be excellent tools for monitoring and evaluating environmental stresses.

Rock n' Seeds: A database of seed functional traits and germination experiments from Brazilian rock outcrop vegetation.

Advancing functional ecology depends fundamentally on the availability of data on reproductive traits, including those from tropical plants, which have been historically underrepresented in global trait databases. Although some valuable databases have been created recently, they are mainly restricted to temperate areas and vegetative traits such as leaf and wood traits. Here, we present Rock n' Seeds, a database of seed functional traits and germination experiments from Brazilian rock outcrop vegetation, recognized as outstanding centers of diversity and endemism. Data were compiled through a systematic literature search, resulting in 103 publications from which seed functional traits were extracted. The database includes information on 16 functional traits for 383 taxa from 148 genera, 50 families, and 25 orders. These 16 traits include two dispersal, six production, four morphological, two biophysical, and two germination traits-the major axes of the seed ecological spectrum. The database also provides raw data for 48 germination experiments, for a total of 10,187 records for 281 taxa. Germination experiments in the database assessed the effect of a wide range of abiotic and biotic factors on germination and different dormancy-breaking treatments. Notably, 8255 of these records include daily germination counts. This input will facilitate synthesizing germination data and using this database for a myriad of ecological questions. Given the variety of seed traits and the extensive germination information made available by this database, we expect it to be a valuable resource advancing comparative functional ecology and guiding seed-based restoration and biodiversity conservation in tropical megadiverse ecosystems. There are no copyright restrictions on the data; please cite this paper when using the current data in publications; also the authors would appreciate notification of how the data are used in publications.

O avanço da ecologia funcional depende fundamentalmente da disponibilidade de dados sobre traços reprodutivos, incluindo dados de plantas tropicais, que têm sido historicamente subrepresentados em bancos de dados de traços funcionais globais. Embora alguns bancos de dados valiosos tenham sido criados recentemente, eles são restritos principalmente a áreas temperadas e a traços vegetativos, como traços de folhas e madeira. Neste artigo apresentamos Rock n' Seeds, um banco de dados de traços funcionais de sementes e experimentos de germinação de vegetações associadas a afloramentos rochosos do Brasil, os quais são reconhecidos como centros notáveis de diversidade e endemismo. Os dados foram compilados através de uma revisão sistemática na literatura, resultando em 103 publicações das quais foram extraídos os traços funcionais das sementes. O banco de dados inclui informações de 16 traços funcionais para 383 taxa de 148 gêneros, 50 famílias e 25 ordens. Estes dezesseis traços incluem dois traços de dispersão, seis de produção, quatro morfológicos, dois biofísicos e dois germinativos; os eixos principais do espectro ecológico da semente. O banco de dados também fornece os dados brutos para 48 experimentos de germinação para um total de 10.187 registros para 281 taxa. Os experimentos de germinação no banco de dados avaliaram o efeito de uma ampla gama de fatores abióticos e bióticos sobre a germinação e diferentes tratamentos de quebra de dormência. Particularmente, 8.255 desses registros incluem a contagem diária da germinação. Estas informações facilitarão a síntese de dados de germinação e a utilização deste banco de dados para uma grande variedade de questões ecológicas. Dada a variedade de traços das sementes e as amplas informações sobre germinação disponibilizadas por este banco de dados, esperamos que ele seja um recurso valioso para o avanço da ecologia funcional comparativa e para orientar a restauração baseada em sementes e a conservação da biodiversidade em ecossistemas tropicais megadiversos. Não há restrições de direitos autorais sobre os dados; favor citar este artigo ao utilizar os dados nas publicações e os autores agradeceriam uma notificação de como os dados são utilizados nas publicações.

El avance de la ecología funcional depende fundamentalmente de la disponibilidad de datos sobre rasgos reproductivos-incluyendo los de las plantas tropicales-los cuales han estado poco representados en las bases de datos globales de rasgos. Aunque recientemente se han creado algunas bases de datos valiosas, estas se encuentran restringidas principalmente a las zonas templadas y a los rasgos vegetativos, como los de las hojas y la madera. En este artículo presentamos Rock n' Seeds, una base de datos de rasgos funcionales de semillas y experimentos de germinación de la vegetación asociada a afloramientos rocosos de Brasil, los cuales son destacados centros de diversidad y endemismo. Los datos se recopilaron mediante una búsqueda bibliográfica sistemática, que dio como resultado 103 publicaciones de las que se extrajeron los rasgos funcionales de las semillas. La base de datos incluye información de dieciséis rasgos funcionales para 383 taxones de 148 géneros, 50 familias y 25 órdenes. Estos rasgos incluyen dos rasgos de dispersión, seis de producción, cuatro morfológicos, dos biofísicos y dos de germinación; siendo estos los principales ejes del espectro ecológico de las semillas. La base de datos también proporciona los datos brutos de 48 experimentos de germinación, para un total de 10.187 registros de 281 taxones. Dichos experimentos de germinación evaluaron el efecto de una amplia gama de factores abióticos y bióticos sobre la germinación y de diferentes tratamientos para romper la dormancia. En particular, 8.255 de estos registros cuentan con conteos diarios de germinación. Esto facilitará la síntesis de los datos de germinación y el uso de esta base de datos para una gran diversidad de preguntas ecológicas. Dada la variedad de rasgos de las semillas y la amplia información sobre germinación que ofrece esta base de datos, esperamos que sea un recurso valioso para el avance de la ecología funcional comparativa y para orientar la restauración basada en semillas y la conservación de la biodiversidad en ecosistemas tropicales megadiversos. No hay restricciones de derechos de autor sobre los datos; se solicita citar este documento cuando se utilicen los datos en publicaciones y los autores agradecerán ser notificados sobre cómo se utilizan los datos en las publicaciones.

Ants on plants: a meta-analysis of the role of ants as plant biotic defenses.

We reviewed the evidence on the role of ants as plant biotic defenses, by conducting meta-analyses for the effects of experimental removal of ants on plant herbivory and fitness with data pooled from 81 studies. Effects reviewed were plant herbivory, herbivore abundance, hemipteran abundance, predator abundance, plant biomass and reproduction in studies where ants were experimentally removed (n = 273 independent comparisons). Ant removal exhibited strong effects on herbivory rates, as plants without ants suffered almost twice as much damage and exhibited 50% more herbivores than plants with ants. Ants also influenced several parameters of plant fitness, as plants without ants suffered a reduction in biomass (-23.7%), leaf production (-51.8%), and reproduction (-24.3%). Effects were much stronger in tropical regions compared to temperate ones. Tropical plants suffered almost threefold higher herbivore damage than plants from temperate regions and exhibited three times more herbivores. Ant removal in tropical plants resulted in a decrease in plant fitness of about 59%, whereas in temperate plants this reduction was not statistically significant. Ant removal effects were also more important in obligate ant-plants (=myrmecophytes) compared to plants exhibiting facultative relationships with hemiptera or those plants with extrafloral nectaries and food bodies. When only tropical plants were considered and the strength of the association between ants and plants taken into account, plants with obligate association with ants exhibited almost four times higher herbivory compared to plants with facultative associations with ants, but similar reductions in plant reproduction. The removal of a single ant species increased plant herbivory by almost three times compared to the removal of several ant species. Altogether, these results suggest that ants do act as plant biotic defenses, but the effects of their presence are more pronounced in tropical systems, especially in myrmecophytic plants.

Constituents of glandular trichomes of Tithonia diversifolia: relationships to herbivory and antifeedant activity.

The herbivory activity of the bordered patch larvae (Chlosyne lacinia, Lepidoptera) on leaves of a Brazilian population of Tithonia diversifolia and the antifeedant potential of its leaf rinse extract were investigated. The caterpillars fed only on the adaxial face, where the density of glandular trichomes is very low, and avoided the abaxial face, which contains high levels of trichomes. Deterrent activity against the larvae was observed in leaf discs treated with leaf rinse extract at concentrations of 1-5% of fresh leaf weight. High-performance liquid chromatography (HPLC) analysis indicated that sesquiterpene lactones are the main constituents of the glandular trichomes. Dichloromethane rinse extracts of the leaves and inflorescences were chemically investigated, and 16 compounds were isolated and identified: 14 sesquiterpene lactones, a flavonoid and a diterpenoid. In this study, five sesquiterpene lactones are described for the first time in the genus, including two lactones, one of which has an unusual seco-guaianolide skeleton. Our findings indicate that the caterpillars avoid the sesquiterpene-lactone-rich glandular trichomes, and provide evidence for the antifeedant activity of the dichloromethane leaf rinse extract. In addition, a study of the seasonal variation of the main constituents from the leaf surface throughout a year demonstrated that a very low qualitative but a very high quantitative variation occurs. The highest level of the main metabolite tagitinin C was observed between September and October and the lowest was from March to June, the later corresponding to the period of highest infestation by the larvae.

Aftershocks of the Samarco disaster: diminished growth and increased metal content of Raphanus sativuscultivated in soil with mining tailings

The collapse of the Fundão tailings dam of the Samarco mining complex in Mariana, Brazil, was the largest mining disaster in the world to date with many socio-economic and environmental impacts. Soil affected by mining tailings was severely altered with negative impacts for agriculture. We tested whether diluting mining tailings with organic soil would eliminate or at least attenuate the ecotoxic effects on plant development and performance. We cultivated radish, Raphanus sativus,in substrates containing different proportions of mining tailings and organic soil: pure tailings (T100%); 2) tailings75%+ soil25%(T75%); 3) tailings50%+ soil50%(T50%); 4) tailings25%+ soil75%(T25%), and 5) pure organic soil (Soil100%, control). There were large differences in soil quality parameters between the 100% tailings treatment (T100%) and the control (Soil100%), as well as for some parameters in the most diluted treatment -T25%(Ca2+, Fe, Mn) in relation to the controltreatment. Although dilution of the tailings soil improved radish development, there was lower radish productivity (leaf area, total biomass, and root/tuber biomass) than for pure soil (control). There were also significantly higher amounts of bioaccumulated metals in radish tubers grown with tailings, even when grown in T25%for Fe content and in T75%for Mn content. These results present a worrisome scenario for human communities in the region of the Doce river, as human consumption of crops produced in soil contaminated with tailingsis not recommended due to potential toxicological effects from high metal concentrations.(AU)

CO2 Fertilizer Effect on Growth, Polyphenols, and Endophytes in Two Baccharis Species

Abstract In a climate change context, the buildup of CO2 will affect plant communities worldwide. This study evaluated the effects of CO2 enrichment on the development and defense of two Cerrado native species Baccharis dracunculifolia and B. platypoda and their associated endophytic fungi richness. The study took place in Open-Top Chambers, two with ambient CO2 concentration (~400 ppm) and two in an enriched environment (~800 ppm). Baccharis platypoda developed 20% more leaves under enriched CO2 conditions, whereas B. dracunculifolia was 30% taller and showed 27% more leaves than those under ambient conditions. In both species, leaf polyphenol concentration did not differ between treatments. Nevertheless, polyphenol content had a positive correlation with plant height on both species' individuals grown under CO2 enriched conditions. Endophytic fungi richness and colonization rate on both plant species did not differ between ambient and enriched conditions. Our results show the positive effect of CO2 fertilizer in at least one of the measured growth parameters. An important new finding was a synergistic increase in growth and chemical defense in both studied species under enriched CO2 conditions, suggesting higher carbon assimilation and accumulation. This study suggests that the effects on primary productivity and secondary metabolites of Baccharis species will potentially reflect on the diversity and distribution of Cerrado plants and their associated animal communities.