Assessing heavy metal contamination in a Brazilian metropolis: a case study with a focus on (bio)indicators.

The continuous expansion of the global vehicle fleet poses a growing threat to environmental quality through heavy metal contamination. In this scenario, monitoring to safeguard public health in urban areas is necessary. Our study involved the collection of 36 street dust and 29 moss samples from roads of a Brazilian metropolis (Recife) with varying traffic intensities as follows: natural reserve (0 vehicles per day), low (< 15,000 vehicles per day), medium (15,000-30,000 vehicles per day), and high (> 30,000 vehicles per day). ICP-AES analysis was performed to determine the concentrations of nine potentially toxic metals (Ba, Cd, Cr, Cu, Mn, Ni, Pb, V, and Zn) to assess the influence of vehicular flow on urban contamination. In the street dust samples, the mean metal concentrations (mg kg-1) exhibited the following order: Ba (503.7) > Mn (303.0) > Zn (144.4) > Cu (95.3) > Cr (56.1) > Pb (34.2) > V (28.7) > Ni (11.3) > Cd (1.5). Conversely, in the moss samples, the metal concentration order was as follows (mg kg-1): Mn (63.8) > Zn (62.5) > Ba (61.0) > Cu (17.7) > Cr (8.0) > V (7.3) > Pb (7.0) > Ni (2.9) > Cd (0.3). Roads with higher traffic volumes exhibited the highest metal enrichments in moss samples for all metals and in dust samples for Cd, Cr, Mn, Ni, and V. However, dust from low-flow roads had higher enrichments for Ba, Cu, and Zn, indicating the influential role of other traffic-related factors in metal deposition. Our findings highlight traffic flow as the predominant source of pollution in urban centers, with both street dust and moss serving as sensitive indicators of metal input attributable to vehicular traffic. These indicators offer valuable insights for urban quality monitoring and pollution control efforts.

First assessment of atmospheric pollution by trace elements and particulate matter after a severe collapse of a tailings dam, Minas Gerais, Brazil: An insight into biomonitoring with Tillandsia usneoides and a public health dataset.

In this study, samples of bromeliad Tillandsia usneoides (n = 70) were transplanted and exposed for 15 and 45 days in 35 outdoor residential areas in Brumadinho (Minas Gerais state, Brazil) after one of the most severe mining dam collapses in the world. Trace elements aluminum (Al), arsenic (As), chromium (Cr), copper (Cu), iron (Fe), mercury (Hg), manganese (Mn), nickel (Ni) and zinc (Zn) were quantified by atomic absorption spectrometry. Scanning electron microscope generated surface images of T. usneoides fragments and particulate matter (PM2.5, PM10 and PM > 10). Aluminum, Fe and Mn stood out from the other elements reflecting the regional geological background. Median concentrations in mg kg-1 increased (p < 0.05) between 15 and 45 days for Cr (0.75), Cu (1.23), Fe (474) and Mn (38.1), while Hg (0.18) was higher at 15 days. The exposed-to-control ratio revealed that As and Hg increased 18.1 and 9.4-fold, respectively, not showing a pattern associated only with the most impacted sites. The PM analysis points to a possible influence of the prevailing west wind on the increase of total particles, PM2.5 and PM10 in transplant sites located to the east. Brazilian public health dataset revealed increase in cases of some cardiovascular and respiratory diseases/symptoms in Brumadinho in the year of the dam collapse (1.38 cases per 1000 inhabitants), while Belo Horizonte capital and its metropolitan region recorded 0.97 and 0.37 cases, respectively. Although many studies have been carried out to assess the consequences of the tailings dam failure, until now atmospheric pollution had not yet been evaluated. Furthermore, based on our exploratory analysis of human health dataset, epidemiological studies are required to verify possible risk factors associated with the increase in hospital admissions in the study area.

Study of the Biological Activity and Phenolic Content of the Ethanol Extract of Phyllogonium viride Brid. (Phyllogoniaceae, Bryophyta).

The present study aimed to examine the phenolic content and evaluate the antimicrobial and antioxidant potential of ethanol extracts from the moss species Phyllogonium viride Brid. on the pathogenic bacteria Salmonella enterica serovar enteritidis, Staphylococcus aureus, Listeria monocytogenes and Escherichia coli, and the pathogenic fungi Candida albicans and Cryptococcus neoformans. The antimicrobial activity was determined from Minimum Inhibitory Concentration (MIC) Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC). Antioxidant activity was determined by the DPPH method. Folin-Denis reagent was used for the content of total phenolics and flavonoids and HPLC-DAD for identification of phenolic compounds. The results showed that bacteriostatic and bactericidal activities occurred at concentrations ranging from 9.76 µg/mL-78.13 µg/mL among all evaluated microorganisms. These values, considering the criteria used, suggest the P. viride extract as a potent antimicrobial. For antioxidant activity, P. viride extract was considered weak. Analysis of the phenolic content showed a wide range of compounds, with Kaempferol (0.41 mg/g) being the major compound, followed by t-cinnamic acid and caffeic acid (0.17 mg/g). Although P. viride is a species of moss not yet referenced in scientific publications of biotechnological interest, it has shown promising potential for further studies and possible application as an antimicrobial of natural origin.

Pseudocrossidium replicatum (Taylor) R.H. Zander is a fully desiccation-tolerant moss that expresses an inducible molecular mechanism in response to severe abiotic stress.

KEY MESSAGE: The moss Pseudocrossidium replicatum is a desiccation-tolerant species that uses an inducible system to withstand severe abiotic stress in both protonemal and gametophore tissues. Desiccation tolerance (DT) is the ability of cells to recover from an air-dried state. Here, the moss Pseudocrossidium replicatum was identified as a fully desiccation-tolerant (FDT) species. Its gametophores rapidly lost more than 90% of their water content when exposed to a low-humidity atmosphere [23% relative humidity (RH)], but abscisic acid (ABA) pretreatment diminished the final water loss after equilibrium was reached. P. replicatum gametophores maintained good maximum photosystem II (PSII) efficiency (Fv/Fm) for up to two hours during slow dehydration; however, ABA pretreatment induced a faster decrease in the Fv/Fm. ABA also induced a faster recovery of the Fv/Fm after rehydration. Protein synthesis inhibitor treatment before dehydration hampered the recovery of the Fv/Fm when the gametophores were rehydrated after desiccation, suggesting the presence of an inducible protective mechanism that is activated in response to abiotic stress. This observation was also supported by accumulation of soluble sugars in gametophores exposed to ABA or NaCl. Exogenous ABA treatment delayed the germination of P. replicatum spores and induced morphological changes in protonemal cells that resembled brachycytes. Transcriptome analyses revealed the presence of an inducible molecular mechanism in P. replicatum protonemata that was activated in response to dehydration. This study is the first RNA-Seq study of the protonemal tissues of an FDT moss. Our results suggest that P. replicatum is an FDT moss equipped with an inducible molecular response that prepares this species for severe abiotic stress and that ABA plays an important role in this response.

The largest moss carpet transplant in Antarctica and its bryosphere cryptic biodiversity.

As part of the reconstruction of the Brazilian Antarctic Station on King George Island, three areas of moss carpet were transplanted to minimize the impact of the new facilities on the local biodiversity. A total of 650 m2 of moss carpet was transplanted to neighboring but previously uncolonized locations and has subsequently survived for the last 3 years. Antarctic moss carpets typically comprise low moss species diversity and are often monospecific. We investigated the cryptic biodiversity that was transplanted along with the carpets using a metabarcoding approach through high throughput sequencing. We targeted 16S rRNA for Bacteria and Archaea, ITS for Fungi and Viridiplantae and Cox1 for Metazoa. We detected DNA representing 263 taxa from five Kingdoms (Chromista, Fungi, Metazoa, Protista and Viridiplantae), two Domains (Archaea and Bacteria) and 33 Phyla associated with the carpet. This diversity included one Archaea, 189 Bacteria, 24 Chromista, 19 Fungi, eight Metazoa, seven Protista and 16 Viridiplantae. Bacteria was the most abundant, rich and diverse group, with Chromista second in diversity and richness. Metazoa was less diverse but second highest in dominance. This is the first study to attempt transplanting a significant area of moss carpet to minimize anthropogenic environmental damage in Antarctica and to use metabarcoding as a proxy to assess diversity associated with Antarctic moss carpets, further highlighting the importance of such habitats for other organisms and their importance for conservation.

Chemical Composition and Cytotoxic Evaluation of the Essential Oil of Phyllogonium viride (Phyllogoniaceae, Bryophyta).

The present study aimed to determine the chemical composition and biological activity of the essential oil obtained from Phyllogonium viride Brid. (Phyllogoniaceae, Bryophyta), whose samples were collected in southern Brazil. For the first time, the cytotoxic activity of the essential oil of P. viride in breast and colorectal tumor cells (MCF-7 and HCT-116) was evaluated, as well as the cytotoxic potential of this oil in non-tumoral cells of human immortalized keratinocytes (HaCaT) via MTT assay. The compounds majorly found in P. viride essential oil were ß-bazzanene (20.30 %), ß-caryophyllene (17.06 %), ß-chamigrene (14.02), and germacrene B (11.72 %). Treatment with P. viride essential oil in the different tested cell lines did not induce any toxicity in most of the tested concentrations. These data contribute to generating new scientific information about this understudied plant species. Furthermore, the chemical characterization of the compounds present in the essential oil of P. viride can lead to greater elucidation of its biotechnological potential.

Gastrotrichs and tardigrades in a remnant of Atlantic Forest (Serra do Japi, SP, Brazil) / Gastrótricos e tardígrados em um remanescente de Mata Atlântica (Serra do Japi, SP, Brasil)

Abstract: Serra do Japi, located in the southeast of São Paulo State, is considered a priority area for conservation, as it houses original Atlantic Forest cover remains. Despite the significant number of studies about vertebrates and invertebrates that were carried out in this region, the meiofauna biodiversity is completely unknown. Thus, the present study aimed to investigate for the first time freshwater Gastrotricha and limnoterrestrial Tardigrada in Serra do Japi Biological Reserve. Samples of sediments, periphyton and floating vegetation in reservoirs and natural lagoons, and mosses growing on native and non-native tree trunks were collected in May 2019. At least five gastrotrichs morphotypes were identified and three of them were formally described: Chaetonotus acanthocephalus, C. dadayi (first record in Brazil), and Heterolepidoderma mariae (first record outside the type locality). In regards to tardigrades, twelve morphotypes were identified and four of them were formally described: Pseudechiniscus juanitae, Minibiotus cf. acontistus, Echiniscus dreyfusi and Itaquascon umbellinae (last two species reported for the first time outside the type locality). This study reinforces that meiofaunal diversity and distribution have been underestimated, even in one of the five largest hotspots in the world.

Resumo: A Serra do Japi, localizada no sudeste do Estado de São Paulo, é considerada uma área prioritária para conservação, por abrigar vestígios de cobertura original de Mata Atlântica. Apesar do número significativo de estudos sobre vertebrados e invertebrados realizados nesta região, a biodiversidade de grupos meiofaunais é completamente desconhecida. Assim, o presente estudo teve como objetivo investigar pela primeira vez Gastrotricha de água doce e Tardigrada limnoterrestre na Reserva Biológica da Serra do Japi. As amostras de sedimentos, perifíton e vegetação flutuante em reservatórios e lagoas naturais, e musgos presentes em troncos de árvores nativas e não nativas foram coletadas em maio de 2019. Foram encontrados cinco morfotipos de gastrótricos, sendo que apenas três deles já foram formalmente descritos: Chaetonotus acanthocephalus, C. dadayi (primeiro registro no Brasil) e Heterolepidoderma mariae (primeiro registro fora da localidade-tipo). Em relação aos tardígrados, foram identificados doze morfotipos, sendo que quatro deles já formalmente descritos: Pseudechiniscus juanitae, Minibiotus cf. acontistus, Echiniscus dreyfusi e Itaquascon umbellinae (as duas últimas espécies relatadas pela primeira vez fora da localidade-tipo). Este trabalho reforça que a diversidade e distribuição da meiofauna têm sido subestimadas, mesmo em um dos cinco maiores hotspots do mundo.

Moss inhabiting flea beetles (Coleoptera: Chrysomelidae: Galerucinae: Alticini) of the West Indies II: Menudos, a new genus from Puerto Rico and description of methods to collect moss inhabiting flea beetles.

Menudos Linzmeier Konstantinov, a new genus of moss inhabiting flea beetles, containing five species, three of them are new (M. illariosus, M. maricao, and M. toronegro-the type species of the genus), from Puerto Rico is described and illustrated. The new genus is compared to Aedmon Clark, Apleuraltica Bechyne, Andersonaltica Linzmeier Konstantinov, Distigmoptera Blake and Ulrica Scherer. Aedmon barberi (Blake, 1943b) and Distigmoptera chamorrae Konstantinov Konstantinova, 2011 are transferred to Menudos. A key to Menudos species identification is provided. Methods for collecting moss inhabiting flea beetles and other arthropods are described in detail for the first time.

Moss inhabiting flea beetles of the West Indies III: Erinaceialtica, a new genus from Hispaniola (Coleoptera, Chrysomelidae, Galerucinae, Alticini).

Erinaceialtica Konstantinov & Linzmeier, a new genus of moss inhabiting flea beetles, containing seven species from the Dominican Republic and Haiti is described and illustrated. Five species are new (E. gabbysalazarae, E. janestanleyae, E. rickstanleyi (the type species), E. rileyi, and E. thomasi) and two species are transferred from Aedmon: A. albicincta (Blake) and A. hugonis (Blake) comb. nov. The new genus is compared to Aedmon Clark, Apleuraltica Bechyne, Andersonaltica Linzmeier & Konstantinov, Distigmoptera Blake, and Ulrica Scherer. Keys to Erinaceialtica and related genera and to Erinaceialtica species are provided.

Active fractions of mannoproteins derived from yeast cell wall stimulate innate and acquired immunity of adult and elderly dogs.

Nutritional intervention in older dogs aims to increase lifespan and improve life quality as well as delay the development of diseases related to ageing. It is believed that active fractions of mannoproteins (AFMs) obtained through extraction and fractionation of yeast cell walls (Saccharomyces cerevisiae) may beneficially modulate the immune system. However, studies that have evaluated this component and the effects of ageing on the immune system of dogs are scarce. This study aimed to evaluate the immunological effects of AFMs in adult and elderly dogs. Three extruded iso-nutrient experimental diets were formulated: without addition of AFM (T0); with AFM at 400 mg/kg (T400); and with AFM at 800 mg/kg (T800). Thirty-six beagle dogs were used, and six experimental treatments, resulting in combinations of age (adult and elderly) and diet (T0, T400, and T800), were evaluated. On days zero, 14, and 28, blood samples were obtained for leucocyte phenotyping and phagocytosis assays. On days zero and 28, a lymphoproliferation test, quantification of reactive oxygen (H2O2) and nitrogen (NO) intermediate production, evaluation of faecal immunoglobulin A (IgA) content, and a delayed cutaneous hypersensitivity test (DCHT) were performed. Statistical analyses were performed with SAS software. Repeated measure variance analyses were performed, and means were compared by the Tukey test. Values of P ≤ 0.05 were considered significant, and values of P ≤ 0.10 were considered tendencies. Dogs fed T400 tended to have higher neutrophilic phagocytic activity than dogs fed T800 (P = 0.073). Regarding reactive oxygen intermediates, bacterial lipopolysaccharide (LPS)-stimulated neutrophils from animals that were fed T400 had a tendency to produce more H2O2 than those from animals fed the control diet (P = 0.093). Elderly dogs, when compared to adult dogs, had lower absolute T and B lymphocyte counts, lower auxiliary T lymphocyte counts, and higher cytotoxic T lymphocyte counts (P < 0.05). A significant effect of diet, age, and time with saline inoculation was noted for the DCHT. There was no effect of diet or age on faecal IgA content in dogs. This study suggests beneficial effects of mannoproteins on the specific and nonspecific immune responses in adult and elderly dogs.

Two new species of Hydraena Kugelann, 1794 from Brazil (Coleoptera: Hydraenidae).

Hydraena (Hydraenopsis) josefinae sp. nov. and H. (H.) pernambucana sp. nov. are described respectively from Amapá and Pernambuco States, Brazil. Hydraena (H.) josefinae sp. nov. is a member of the scintillabella subgroup of the leechi group, while H. (H.) pernambucana sp. nov. belongs to the orcula complex in the marginicollis subgroup of the marginicollis group. Descriptions and diagnostic characters for both new species are offered; habitus photographs and line drawings of male genitalia of the two new species, as well as drawings of female genitalia of H. josefinae sp. n., are provided. Male genitalia of H. alterra Perkins, 1980 is illustrated and compared with that of the closely related H. josefinae sp. nov. Habitat and distributional data of the two new species are also included.

Growth supression of plant pathogenic fungi using bryophite extracts / Supressão de crescimento de fungos patogênicos de plantas usando extratos de briófitas

Chemicals are often used in attempts to control diseases caused by plant pathogenic fungi during food production. However, chemicals can have adverse effects not just on food, but they also remain active for a long time within ecosystems, and thus are not environmentally friendly. Therefore,development of bio-treatment and avoiding use of chemicals are urgently needed. With the aim of studying and developing new environmentally-friendly treatments, we tested extracts from selected bryophyte species(Porella platyphylla, Cinclidotus fontinaloides and Anomodon viticulosus) on five plant pathogenic fungi under controlled conditions. The fungi (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum and Monilinia laxa) were selected based on common diseases they cause on fruits and grapevine. They were isolated in cultures and treated with bryophyte extracts. Bryophyte extracts were shown to be effective in suppression of certain plant pathogenic fungi growth and to have a huge potential in development of novel biotechnological treatments and biofungicides. The best results were achieved in inhibition of B. dothidea, P. viticola and Calosphaeria sp.

Fungos fitopatogênicos são controlados com produtos químicos para combater doenças causadas por eles durante a produção de alimentos. Tais produtos são ruins não apenas para a alimentação, mas também podem permanecer por muito tempo nos ecossistemas, portanto, não são ecologicamente corretos. Desta forma, os biotratamentos e a prevenção de usos químicos são urgentemente necessários no futuro próximo. Com o objetivo de estudar e desenvolver nova alternative decontrole de doenças de plantas, testamos os extratos de espécies de briófitas selecionadas (Porella platyphylla, Cinclidotus fontinaloides e Anomodon viticulosus) em cinco fungos fitopatogênicos (Botryosphaeria dothidea, Phomopsis viticola, Calosphaeria sp., Colletotrichum acutatum e Monilinia laxa) em condição controlada. Estes fungos, selecionados com base nas doenças que causam em frutos e videiras, foram isolados em culturas puras e tratados com os respectivos extratos de briófitas. Os extratos de briófitas demonstraram ser eficazes na supressão de certos fungos fitopatogênicos e têm um enorme potencial no desenvolvimento de novos biofungicidas e tratamentos biotecnológicos. Os mais interessantes resultados foram obtidos na inibição de B. dothidea, P. viticola eCalosphaeria sp.

Photoprotection enhanced by red cell wall pigments in three East Antarctic mosses.

BACKGROUND: Antarctic bryophytes (mosses and liverworts) are resilient to physiologically extreme environmental conditions including elevated levels of ultraviolet (UV) radiation due to depletion of stratospheric ozone. Many Antarctic bryophytes synthesise UV-B-absorbing compounds (UVAC) that are localised in their cells and cell walls, a location that is rarely investigated for UVAC in plants. This study compares the concentrations and localisation of intracellular and cell wall UVAC in Antarctic Ceratodon purpureus, Bryum pseudotriquetrum and Schistidium antarctici from the Windmill Islands, East Antarctica. RESULTS: Multiple stresses, including desiccation and naturally high UV and visible light, seemed to enhance the incorporation of total UVAC including red pigments in the cell walls of all three Antarctic species analysed. The red growth form of C. purpureus had significantly higher levels of cell wall bound and lower intracellular UVAC concentrations than its nearby green form. Microscopic and spectroscopic analyses showed that the red colouration in this species was associated with the cell wall and that these red cell walls contained less pectin and phenolic esters than the green form. All three moss species showed a natural increase in cell wall UVAC content during the growing season and a decline in these compounds in new tissue grown under less stressful conditions in the laboratory. CONCLUSIONS: UVAC and red pigments are tightly bound to the cell wall and likely have a long-term protective role in Antarctic bryophytes. Although the identity of these red pigments remains unknown, our study demonstrates the importance of investigating cell wall UVAC in plants and contributes to our current understanding of UV-protective strategies employed by particular Antarctic bryophytes. Studies such as these provide clues to how these plants survive in such extreme habitats and are helpful in predicting future survival of the species studied.

Six new and one known species of Geomonhystera (Nematoda, Monhysteridae) from moss, an epiphytic plant and soil in México and Ecuador.

Six new species of Geomonhystera (Nematoda, Monhysterida: Monhysteridae) Andrássy, 1981, are described and illustrated, five from México and one from Ecuador. The new species have many features in common and in common with at least several other species of the genus. Consequently, those features individually are not very useful for species-level diagnosis. They include the cuticular striation which, under light microscopy, ranges from fine to apparently smooth; however, scanning electron micrographs reveal very fine cuticular striation on all the new species. All the new species have a few body setae scattered along the length of the body and the vulval lips of females are slightly protruding. As for other species of the genus, there is considerable overlap in the ranges of standard morphometric characters so a combination of standard and additional morphometrics, as well as qualitative characters, is necessary for species separation. Geomonhystera mexiquense sp. n., collected from moss on a rock and on the trunk of Quercus crassipes in Juchitepec, México State, México, has outer labial setae that are unsegmented and 44-67% of the head width, the cephalic setae are thin, unsegmented and 31-50% of the head width; the vulval lips protrude slightly. Geomonhystera michoacana sp. n., collected from moss on a rock in Tzararacua National Park, Uruapan, Michoacan State, México, has a very finely-striated cuticle which appears almost smooth and bears rare cervical and caudal setae; the tail is long, 15-17% of the body length, and the rectum also is long, 23-32 (27±2.0) µm. Geomonhystera longispiculata sp. n., collected from an epiphytic plant in the botanical garden of the Ecology Institute in Jalapa, Veracruz State, México, is distinguished from all the species of the genus by the long, slender spicules, 55-85 (75±3.0) µm. The cuticle has conspicuous fine striation and there are somatic setae scattered along the body. Geomonhystera ecuatoriana sp. n., collected from moss on the trunk of Scalesia pedunculata growing on the twin volcanoes of Isla Santa Cruz, Galapagos, Ecuador, has fine cuticular striation and a few small body setae; the unsegmented outer labial setae are 42-54% of the head width and the cephalic setae are thin, unsegmented and 33-45% of the head width; the vulval lips protrude slightly and the vulva-anus distance is 37-48 µm or 2.1-2.5 times anal body diameter. Males of G. ecuatoriana have slightly arcuate spicules, 35 µm long. Geomonhystera galindoi sp. n., collected from moss on the trunk of Quercus peduncularis in San Pablo Ixzayo, Texcoco, México State, México, has fine cuticlar striation and a few fine body setae, the vulval lips are slightly protruding; the vulva-anus distance is 35-50 (41±0.9) µm, 1.5-2.2 (1.9±0.2) times anal body diameter. Males of G. galindoi are slightly ventrally curved with the posterior end strongly curved, giving the body a hook-shaped appearance. Geomonhystera chiautzingoensis sp. n., collected from moss on the trunk of Crataegus mexicana in Chiautzingo, Puebla State, México, has exceedingly fine striation of the cuticle and outer labial and cephalic setae that are less than 50% of the head width. Additionally, G. dubia Siddiqi Shahina, 2004 was sollected in soil samples around a plum tree (Prunus sp.) in the garden of Montecillo Campus, Colegio de Postgraduados, Texcoco, México State, México This species has a small body with fine cuticular striation and a few sparsely-distributed body setae; the outer labial and cephalic setae are unsegmented, 5-8 and 3-5 µm long, 56-80% and 33-50% of the head width, respectively; the vulval lips protrude slightly but are sometimes flush with the body contour.

Photoprotection enhanced by red cell wall pigments in three East Antarctic mosses

BACKGROUND: Antarctic bryophytes (mosses and liverworts) are resilient to physiologically extreme environmental conditions including elevated levels of ultraviolet (UV) radiation due to depletion of stratospheric ozone. Many Antarctic bryophytes synthesise UV-B-absorbing compounds (UVAC) that are localised in their cells and cell walls, a location that is rarely investigated for UVAC in plants. This study compares the concentrations and localisation of intracellular and cell wall UVAC in Antarctic Ceratodon purpureus, Bryum pseudotriquetrum and Schistidium antarctici from the Windmill Islands, East Antarctica. RESULTS: Multiple stresses, including desiccation and naturally high UV and visible light, seemed to enhance the incorporation of total UVAC including red pigments in the cell walls of all three Antarctic species analysed. The red growth form of C. purpureus had significantly higher levels of cell wall bound and lower intracellular UVAC concentrations than its nearby green form. Microscopic and spectroscopic analyses showed that the red colouration in this species was associated with the cell wall and that these red cell walls contained less pectin and phenolic esters than the green form. All three moss species showed a natural increase in cell wall UVAC content during the growing season and a decline in these compounds in new tissue grown under less stressful conditions in the laboratory. CONCLUSIONS: UVAC and red pigments are tightly bound to the cell wall and likely have a long-term protective role in Antarctic bryophytes. Although the identity of these red pigments remains unknown, our study demonstrates the importance of investigating cell wall UVAC in plants and contributes to our current understanding of UV-protective strategies employed by particular Antarctic bryophytes. Studies such as these provide clues to how these plants survive in such extreme habitats and are helpful in predicting future survival of the species studied.

Heterologous protein production in the moss Physcomitrella patens

Over the last decades plants have been used for the heterologous production of pharmaceuticals, industrial enzymes and edible vaccines. The moss Physcomitrella patens is considered as an experimental model of choice for basic molecular, cytological and developmental questions in plant biology, as well as an outstanding plant model system for heterologous protein production. Here we use P. patens to produce osmotin, a tobacco protein with fungicidal properties. We have generated a transgenic plant able to synthesize and secrete a biologically active osmotin protein(AU)

Durante las últimas décadas las plantas han sido utilizadas para la producción heteróloga de farmacéuticos, enzimas de uso industrial y vacunas. El musgo Physcomitrella patens es considerado como un modelo de experimentación de elección para abordar preguntas en las áreas de biología molecular, citología y de biología del desarrollo en plantas; así como un excelente sistema modelo para la producción de proteínas heterólogas. En este trabajo se utilizó P. patens para la producción de osmotina, una proteína de tabaco con propiedades fungicidas. Se generó una planta transgénica capaz de sintetizar y secretar osmotina biológicamente activa(AU)

Adaptation Mechanisms in the Evolution of Moss Defenses to Microbes.

Bryophytes, including mosses, liverworts and hornworts are early land plants that have evolved key adaptation mechanisms to cope with abiotic stresses and microorganisms. Microbial symbioses facilitated plant colonization of land by enhancing nutrient uptake leading to improved plant growth and fitness. In addition, early land plants acquired novel defense mechanisms to protect plant tissues from pre-existing microbial pathogens. Due to its evolutionary stage linking unicellular green algae to vascular plants, the non-vascular moss Physcomitrella patens is an interesting organism to explore the adaptation mechanisms developed in the evolution of plant defenses to microbes. Cellular and biochemical approaches, gene expression profiles, and functional analysis of genes by targeted gene disruption have revealed that several defense mechanisms against microbial pathogens are conserved between mosses and flowering plants. P. patens perceives pathogen associated molecular patterns by plasma membrane receptor(s) and transduces the signal through a MAP kinase (MAPK) cascade leading to the activation of cell wall associated defenses and expression of genes that encode proteins with different roles in plant resistance. After pathogen assault, P. patens also activates the production of ROS, induces a HR-like reaction and increases levels of some hormones. Furthermore, alternative metabolic pathways are present in P. patens leading to the production of a distinct metabolic scenario than flowering plants that could contribute to defense. P. patens has acquired genes by horizontal transfer from prokaryotes and fungi, and some of them could represent adaptive benefits for resistance to biotic stress. In this review, the current knowledge related to the evolution of plant defense responses against pathogens will be discussed, focusing on the latest advances made in the model plant P. patens.

HLBT-100: a highly potent anti-cancer flavanone from Tillandsia recurvata (L.) L.

BACKGROUND: The incidence and mortalities from cancers remain on the rise worldwide. Despite significant efforts to discover and develop novel anticancer agents, many cancers remain in the unmet need category. As such, efforts to discover and develop new and more effective and less toxic agents against cancer remain a top global priority. Our drug discovery approach is natural products based with a focus on plants. Tillandsia recurvata (L.) L. is one of the plants selected by our research team for further studies based on previous bioactivity findings on the anticancer activity of this plant. METHODS: The plant biomass was extracted using supercritical fluid extraction technology with CO2 as the mobile phase. Bioactivity guided isolation was achieved by use of chromatographic technics combined with anti-proliferative assays to determine the active fraction and subsequently the pure compound. Following in house screening, the identified molecule was submitted to the US National Cancer Institute for screening on the NCI60 cell line panel using standard protocols. Effect of HLBT-100 on apoptosis, caspase 3/7, cell cycle and DNA fragmentation were assessed using standard protocols. Antiangiogenic activity was carried out using the ex vivo rat aortic ring assay. RESULTS: A flavonoid of the flavanone class was isolated from T. recurvata (L.) L. with potent anticancer activity. The molecule was code named as HLBT-100 (also referred to as HLBT-001). The compound inhibited brain cancer (U87 MG), breast cancer (MDA-MB231), leukemia (MV4-11), melanoma (A375), and neuroblastoma (IMR-32) with IC50 concentrations of 0.054, 0.030, 0.024, 0.003 and 0.05 µM, respectively. The molecule also exhibited broad anticancer activity in the NCI60 panel inhibiting especially hematological, colon, CNS, melanoma, ovarian, breast and prostate cancers. Twenty-three of the NCI60 cell lines were inhibited with GI50 values <0.100 µM. In terms of potential mechanisms of action, the molecule demonstrated effect on the cell cycle as evidenced by the accumulation of cells with 

Prezygotic resource-allocation dynamics and reproductive trade-offs in Calymperaceae (Bryophyta).

PREMISE OF THE STUDY: Resource allocation is difficult to characterize in plants because of the challenges of quantifying gametes and propagules. We surveyed six sympatric, unisexual species in the family Calymperaceae (Bryophyta) to test for trade-offs in prezygotic sexual and asexual expression and density-dependent survivorship of female gametangia. METHODS: We tallied gametangial and asexual propagule output for 1820 shoots from 17 populations of six species at monthly intervals during one year (2010-2011) in a central Amazonian forest. Generalized linear mixed models were used to test for trade-offs in sexual and asexual expression and density-dependent senescence probability of gametangia. Precipitation and microsite variables were also included in the model. KEY RESULTS: For all species, sexual and asexual expression were positively correlated with mean monthly precipitation. Asexually expressing shoots produced significantly fewer gametangia than nonexpressing ones, and the probability of senescence increased with shoot density. Archegonium density per shoot was also consistently lower than the modeled optimum to maximize the number of receptive archegonia. CONCLUSIONS: Trade-offs among reproductive strategies and positive density-dependent senescence of female gametangia suggest that prezygotic sexual and asexual expression come at a tangible investment. However, the apparently inefficient resource-allocation dynamics in the production of female gametangia makes the possible advantages of squandering such investments unclear. One possibility is that the study populations, like those of many dioicous mosses, are skewed toward expressing females with low sporophyte production, which would suggest that asexual reproduction predominates and upstages efficient resource allocation in prezygotic investment.

Oxylipins in moss development and defense.

Oxylipins are oxygenated fatty acids that participate in plant development and defense against pathogen infection, insects, and wounding. Initial oxygenation of substrate fatty acids is mainly catalyzed by lipoxygenases (LOXs) and α-dioxygenases but can also take place non-enzymatically by autoxidation or singlet oxygen-dependent reactions. The resulting hydroperoxides are further metabolized by secondary enzymes to produce a large variety of compounds, including the hormone jasmonic acid (JA) and short-chain green leaf volatiles. In flowering plants, which lack arachidonic acid, oxylipins are produced mainly from oxidation of polyunsaturated C18 fatty acids, notably linolenic and linoleic acids. Algae and mosses in addition possess polyunsaturated C20 fatty acids including arachidonic and eicosapentaenoic acids, which can also be oxidized by LOXs and transformed into bioactive compounds. Mosses are phylogenetically placed between unicellular green algae and flowering plants, allowing evolutionary studies of the different oxylipin pathways. During the last years the moss Physcomitrella patens has become an attractive model plant for understanding oxylipin biosynthesis and diversity. In addition to the advantageous evolutionary position, functional studies of the different oxylipin-forming enzymes can be performed in this moss by targeted gene disruption or single point mutations by means of homologous recombination. Biochemical characterization of several oxylipin-producing enzymes and oxylipin profiling in P. patens reveal the presence of a wider range of oxylipins compared to flowering plants, including C18 as well as C20-derived oxylipins. Surprisingly, one of the most active oxylipins in plants, JA, is not synthesized in this moss. In this review, we present an overview of oxylipins produced in mosses and discuss the current knowledge related to the involvement of oxylipin-producing enzymes and their products in moss development and defense.