Insights into agar and secondary metabolite pathways from the genome of the red alga Gracilaria domingensis (Rhodophyta, Gracilariales).

Gracilariales is a clade of florideophycean red macroalgae known for being the main source of agar. We present a de novo genome assembly and annotation of Gracilaria domingensis, an agarophyte alga with flattened thallus widely distributed along Central and South American Atlantic intertidal zones. In addition to structural analysis, an organizational comparison was done with other Rhodophyta genomes. The nuclear genome has 78 Mbp, with 11,437 predicted coding genes, 4,075 of which did not have hits in sequence databases. We also predicted 1,567 noncoding RNAs, distributed in 14 classes. The plastid and mitochondrion genome structures were also obtained. Genes related to agar synthesis were identified. Genes for type II galactose sulfurylases could not be found. Genes related to ascorbate synthesis were found. These results suggest an intricate connection of cell wall polysaccharide synthesis and the redox systems through the use of L-galactose in Rhodophyta. The genome of G. domingensis should be valuable to phycological and aquacultural research, as it is the first tropical and Western Atlantic red macroalgal genome to be sequenced.

Epidemiologic profile and histopathological findings in Neotropical Primates during and after the yellow fever outbreak in Espírito Santo, Brazil.

Yellow fever (YF) is a viral disease whose transmission involves non-human primates (NHP), mosquitoes, and humans. Between 2016 and 2018 occurred the largest YF outbreak in the last 100 years in Brazil. We analyzed epidemiologic profile and geographic distribution of epizootics and described most frequent histopathological findings in NHP that died during YF outbreak in the state of Espírito Santo. We consider 487 epizootics notifications registered at the State Health Department from January 2017 to July 2020. Throughout the state, 51 (65.4%) municipalities reported epizootics, with more cases in central and metropolitan areas. Reverse transcription polymerase chain reaction and immunohistochemistry were laboratory tests performed for diagnosis of yellow fever, with 160 (32.9%) positive results, 314 (64.5%) negative and 13 (2.7%) inconclusive. Histopathological findings were compared statistically between positive and negative animals for YF. The liver was the most affected organ. Hemorrhage, hepatocyte necrosis, steatosis, cholestasis and eosinophilic degeneration were statistically more frequent in positive animals. Tubular necrosis, nephritis, congestion and lymphoid hypoplasia on spleen were statistically correlated to positive animals. Knowledge of pathogenic aspect of YF is necessary to guarantee that samples from Neotropical primates are properly used for YF surveillance purposes, to ensure appropriate diagnoses and subsequent public health responses.

Phylogenomics, divergence time estimation and trait evolution provide a new look into the Gracilariales (Rhodophyta).

The Gracilariales is a highly diverse, widely distributed order of red algae (Rhodophyta) that forms a well-supported clade. Aside from their ecological importance, species of Gracilariales provide important sources of agarans and possess bioactive compounds with medicinal and pharmaceutical use. Recent phylogenetic analyses from a small number of genes have greatly advanced our knowledge of evolutionary relationships in this clade, yet several key nodes were not especially well resolved. We assembled a phylogenomic data set containing 79 nuclear genes, 195 plastid genes, and 24 mitochondrial genes from species representing all three major Gracilariales lineages, including: Melanthalia, Gracilariopsis, and Gracilaria sensu lato. This data set leads to a fully-resolved phylogeny of Gracilariales, which is highly-consistent across genomic compartments. In agreement with previous findings, Melanthalia obtusata was sister to a clade including Gracilaria s.l. and Gracilariopsis, which were each resolved as well-supported clades. Our results also clarified the long-standing uncertainty about relationships in Gracilaria s.l., not resolved in single and multi-genes approaches. We further characterized the divergence time, organellar genome architecture, and morphological trait evolution in Gracilarales to better facilitate its taxonomic treatment. Gracilariopsis and Gracilaria s.l. are comparable taxonomic ranks, based on the overlapping time range of their divergence. The genomic structure of plastid and mitochondria is highly conserved within each clade but differs slightly among these clades in gene contents. For example, the plastid gene petP is lost in Gracilaria s.l. and the mitochondrial gene trnH is in different positions in the genome of Gracilariopsis and Gracilaria s.l. Our analyses of ancestral character evolution provide evidence that the main characters used to delimitate genera in Gracilariales, such as spermatangia type and features of the cystocarp's anatomy, overlap in subclades of Gracilaria s.l. We discuss the taxonomy of Gracilariales in light of these results and propose an objective and practical classification, which is in agreement with the criteria of monophyly, exclusive characters, predictability and nomenclatural stability.

Dual influence of terrestrial and marine historical processes on the phylogeography of the Brazilian intertidal red alga Gracilaria caudata.

In this study, we explored how past terrestrial and marine climate changes have interacted to shape the phylogeographic patterns of the intertidal red seaweed Gracilaria caudata, an economically important species exploited for agar production in the Brazilian north-east. Seven sites were sampled along the north-east tropical and south-east sub-tropical Brazilian coast. The genetic diversity and structure of G. caudata was inferred using a combination of mitochondrial (COI and cox2-3), chloroplast (rbcL) and 15 nuclear microsatellite markers. A remarkable congruence between nuclear, mitochondrial and chloroplast data revealed clear separation between the north-east (from 03° S to 08° S) and the south-east (from 20° S to 23° S) coast of Brazil. These two clades differ in their demographic histories, with signatures of recent demographic expansions in the north-east and divergent populations in the south-east, suggesting the maintenance of several refugia during the last glacial maximum due to sea-level rise and fall. The Bahia region (around 12° S) occupies an intermediate position between both clades. Microsatellites and mtDNA markers showed additional levels of genetic structure within each sampled site located south of Bahia. The separation between the two main groups in G. caudata is likely recent, probably occurring during the Quaternary glacial cycles. The genetic breaks are concordant with (i) those separating terrestrial refugia, (ii) major river outflows and (iii) frontiers between tropical and subtropical regions. Taken together with previously published eco-physiological studies that showed differences in the physiological performance of the strains from distinct locations, these results suggest that the divergent clades in G. caudata correspond to distinct ecotypes in the process of incipient speciation and thus should be considered for the management policy of this commercially important species.

Organellar genomics: a useful tool to study evolutionary relationships and molecular evolution in Gracilariaceae (Rhodophyta).

Gracilariaceae has a worldwide distribution including numerous economically important species. We applied high-throughput sequencing to obtain organellar genomes (mitochondria and chloroplast) from 10 species of Gracilariaceae and, combined with published genomes, to infer phylogenies and compare genome architecture among species representing main lineages. We obtained similar topologies between chloroplast and mitochondrial genomes phylogenies. However, the chloroplast phylogeny was better resolved with full support. In this phylogeny, Melanthalia intermedia is sister to a monophyletic clade including Gracilaria and Gracilariopsis, which were both resolved as monophyletic genera. Mitochondrial and chloroplast genomes were highly conserved in gene synteny, and variation mainly occurred in regions where insertions of plasmid-derived sequences (PDS) were found. In mitochondrial genomes, PDS insertions were observed in two regions where the transcription direction changes: between the genes cob and trnL, and trnA and trnN. In chloroplast genomes, PDS insertions were in different positions, but generally found between psdD and rrs genes. Gracilariaceae is a good model system to study the impact of PDS in genome evolution due to the frequent presence of these insertions in organellar genomes. Furthermore, the bacterial leuC/leuD operon was found in chloroplast genomes of Gracilaria tenuistipitata, G. chilensis, and M. intermedia, and in extrachromosomal plasmid of G. vermiculophylla. Phylogenetic trees show two different origins of leuC/leuD: genes found in chloroplast and plasmid were placed with proteobacteria, and genes encoded in the nucleus were close to Viridiplantae and cyanobacteria.

Progression of gray matter atrophy in seizure-free patients with temporal lobe epilepsy.

OBJECTIVES: To investigate the presence and progression of gray matter (GM) reduction in seizure-free patients with temporal lobe epilepsy (TLE). METHODS: We enrolled 39 consecutive TLE patients, seizure-free for at least 2 years--20 with magnetic resonance imaging (MRI) signs of hippocampal sclerosis (TLE-HS), 19 with normal MRI (TLE-NL), and 74 healthy controls. For longitudinal analysis, we included individuals who had a second MRI with minimum interval of 18 months: 21 patients (10 TLE-HS, 11 TLE-NL) and 11 controls. Three-dimensional (3D) T1 -weighted images acquired in 3 Tesla MRI were analyzed with voxel-based morphometry (VBM). The images of patients with right-sided interictal epileptogenic zone (EZ) were right-left flipped, as well as a comparable proportion of controls. Cross-sectional analysis: The patients' images from each group were compared to controls to investigate differences in GM volumes. Longitudinal analysis: The first and second images were compared in each group to look for decreased GM volume. RESULTS: Cross-sectional analysis: Patients with TLE-HS had diffuse GM atrophy, including hippocampus and parahippocampal gyrus, insula, frontal, and occipital lobes ipsilateral to EZ, bilateral thalamus and contralateral orbitofrontal gyrus, and caudate. In contrast, TLE-NL group did not present significant differences compared to controls. Longitudinal analysis: TLE-HS presented progressive GM reduction in ipsilateral insula and occipital lobe, contralateral motor area, and bilateral temporal and frontal lobes. TLE-NL had GM progression in ipsilateral hypothalamus and parietal lobe, contralateral cerebellum, and bilateral temporal lobe. Controls did not show changes in GM volume between MRIs. SIGNIFICANCE: Diffuse extrahippocampal GM atrophy is present in seizure-free patients with TLE-HS. In addition, there is progressive GM atrophy in patients with and without HS. These results demonstrate that not only ongoing seizures are involved in the progression of GM atrophy. An underlying pathologic mechanism could be responsible for progressive brain volume loss in TLE patients even in seizure-free periods.

Delimitating cryptic species in the Gracilaria domingensis complex (Gracilariaceae, Rhodophyta) using molecular and morphological data.

Species in the genus Gracilaria that display conspicuously flattened vegetative morphologies are a taxonomically challenging group of marine benthic red algae. This is a result of their species richness, morphological similarity, and broad phenotypic plasticity. Within this group, the Gracilaria domingensis complex is one of the most common, conspicuous, and morphologically variable species along the tropical western Atlantic Ocean. Previous research has identified that members of this complex belong to two distantly related clades. However, despite this increased phylogentic resolution, species delimitations within each of these clades remain unclear. Our study assessed the species diversity within this difficult complex using morphological and molecular data from three genetic markers (cox1, UPA, and rbcL). We additionally applied six single-marker species delimitation methods (SDM: ABGD, GMYCs, GMYCm, SPN, bPTP, and PTP) to rbcL, which were largely in agreement regarding species delimitation. These results, combined with our analysis of morphology, indicate that the G. domingensis complex includes seven distinct species, each of which are not all most closely related: G. cervicornis; a ressurected G. ferox; G. apiculata subsp. apiculata; a new species, Gracilaria baiana sp. nov.; G. intermedia subsp. intermedia; G. venezuelensis; and G. domingensis sensu stricto, which includes the later heterotypic synonym, G. yoneshigueana. Our study demonstrates the value of multipronged strategies, including the use of both molecular and morphological approaches, to decipher cryptic species of red algae.

Phylogeny of Gracilariaceae (Rhodophyta): evidence from plastid and mitochondrial nucleotide sequences.

Gracilariaceae are mostly pantropical red algae and include ~230 species in seven genera. Infrafamilial classification of the group has long been based on reproductive characters, but previous phylogenies have shown that traditionally circumscribed groups are not monophyletic. We performed phylogenetic analyses using two plastid (universal plastid amplicon and rbcL) and one mitochondrial (cox1) loci from a greatly expanded number of taxa to better assess generic relationships and understand patterns of character distributions. Our analyses produce the most well-supported phylogeny of the family to date, and indicate that key characteristics of spermatangia and cystocarp type do not delineate genera as commonly suggested. Our results further indicate that Hydropuntia is not monophyletic. Given their morphological overlap with closely related members of Gracilaria, we propose that Hydropuntia be synonymized with the former. Our results additionally expand the known ranges of several Gracilariaceae species to include Brazil. Lastly, we demonstrate that the recently described Gracilaria yoneshigueana should be synonymized as G. domingensis based on morphological and molecular characters. These results demonstrate the utility of DNA barcoding for understanding poorly known and fragmentary materials of cryptic red algae.

Zinc oxide nanoparticle-reinforced pectin/starch functionalized films: A sustainable solution for biodegradable packaging.

Environmental pollution caused by non-biodegradable plastic pollutants adversely affects various ecosystems. This study proposes the development of novel functional and biodegradable films based on corn starch (CST) and pectin (PEC) containing zinc oxide nanoparticles (ZnONPs) from the casting method. The films exhibited processability, transparency, low water vapor permeation, and desirable mechanical properties for food packaging and coating applications. The ZnONPs acted as a plasticizer, enhancing the film elongation at the break, increasing the pec25-1 (PEC 25 wt% and ZnONPs 1 wt%) elongation from 79.85 to 162.32 %. The improved film elasticity supported by ZnONPs reduced the material stiffness. However, the films still demonstrated an average tensile strength (0.69 MPa) 17-fold higher than the tensile strength (0.04 MPa) of the non-biodegradable commercial film based on poly(vinyl chloride). Furthermore, the ZnONPs enhanced the UV-blocking capabilities of the films, leading to wettable materials with water contact angles lower than 90°. The films showed high biodegradation rates under natural disposal conditions. The results indicated that the pec25-1/ZnONPs film is a promising eco-friendly coating in food preservation due to its biodegradability, suitable mechanical properties, low water vapor permeability, and UV-blocking properties.

Factors associated with caregiver burden of adults with epilepsy in a middle-income country.

PURPOSE: Despite the relevance of socio-economic background in burden of caregivers of persons with epilepsy (PWE), there is a dearth of knowledge about this topic in low-resource settings. Here we attempt to identify the extent of the burden in caregivers of adults with epilepsy in a middle-income country and the factors associated with it. METHODS: In this cross-sectional study, we evaluated the level of burden with the Zarit Caregiver Burden Inventory. A comprehensive questionnaire was developed to assess the demographics of patients and caregivers. Epilepsy-related factors were obtained by interview and file review. Depressive, anxiety symptoms, and sleep quality were evaluated by Beck Depression Inventory, State and Trait Anxiety Inventory, and Pittsburgh Sleep Quality Index. Stigma was evaluated by the Stigma Scale of Epilepsy. RESULTS: The caregiver's burden was mild to moderate in our sample. The factors most strongly associated with the higher caregiver burden were age-onset of epilepsy, PWE years of schooling, needs for assistance in daily life, disabilities, and caregiver's bad-quality of sleep and higher depressive and anxiety symptoms. CONCLUSION: Caregivers of PWE deserve special attention regarding mental health and other health aspects, such as sleep quality.

Group I introns and associated homing endonuclease genes reveals a clinal structure for Porphyra spiralis var. amplifolia (Bangiales, Rhodophyta) along the Eastern coast of South America.

BACKGROUND: Group I introns are found in the nuclear small subunit ribosomal RNA gene (SSU rDNA) of some species of the genus Porphyra (Bangiales, Rhodophyta). Size polymorphisms in group I introns has been interpreted as the result of the degeneration of homing endonuclease genes (HEG) inserted in peripheral loops of intron paired elements. In this study, intron size polymorphisms were characterized for different Porphyra spiralis var. amplifolia (PSA) populations on the Southern Brazilian coast, and were used to infer genetic relationships and genetic structure of these PSA populations, in addition to cox2-3 and rbcL-S regions. Introns of different sizes were tested qualitatively for in vitro self-splicing. RESULTS: Five intron size polymorphisms within 17 haplotypes were obtained from 80 individuals representing eight localities along the distribution of PSA in the Eastern coast of South America. In order to infer genetic structure and genetic relationships of PSA, these polymorphisms and haplotypes were used as markers for pairwise Fst analyses, Mantel's test and median joining network. The five cox2-3 haplotypes and the unique rbcL-S haplotype were used as markers for summary statistics, neutrality tests Tajima's D and Fu's Fs and for median joining network analyses. An event of demographic expansion from a population with low effective number, followed by a pattern of isolation by distance was obtained for PSA populations with the three analyses. In vitro experiments have shown that introns of different lengths were able to self-splice from pre-RNA transcripts. CONCLUSION: The findings indicated that degenerated HEGs are reminiscent of the presence of a full-length and functional HEG, once fixed for PSA populations. The cline of HEG degeneration determined the pattern of isolation by distance. Analyses with the other markers indicated an event of demographic expansion from a population with low effective number. The different degrees of degeneration of the HEG do not refrain intron self-splicing. To our knowledge, this was the first study to address intraspecific evolutionary history of a nuclear group I intron; to use nuclear, mitochondrial and chloroplast DNA for population level analyses of Porphyra; and intron size polymorphism as a marker for population genetics.

Marine and coastal biodiversity studies, 60 years of research funding from FAPESP, what we have learned and future challenges / Estudos de biodiversidade marinha e costeira, 60 anos de financiamento à pesquisa da FAPESP, o que aprendemos e desafios futuros

Abstract In this study we survey and analyze 300 projects related to marine biodiversity funded by FAPESP from 1972 to 2021, of which 46 were nested in the BIOTA Program. From a unique project in the 1970's, the number gradually increased until 2009, when BIOTA promoted a call on marine biodiversity, which led to a boost in the number of funded projects in the subsequent years. The geographical range of the projects expanded over the years and, from studies based on the coast of São Paulo State, the focus gradually shifted to broader areas of the Brazilian coast, then to other areas of the Atlantic, and eventually became global. The majority of projects focused on coastal benthic organisms living on hard-bottom. In terms of taxa, six groups accounted for about 60% of the projects (viz. Crustacea, Actinopterygii, Mollusca, Chondrichthyes, Cnidaria, and Rhodophyta), but it is observed an increase in the number of groups studied over the decades. The 300 projects refer to a set of 82 different topics, of which the top five are taxonomy, phylogeny, community, "omics", and pollution. The analyses show a long-standing effort in marine biodiversity surveys, with ongoing updated approaches regarding scope and methods. Research on strategic areas is discussed, including deep-sea and marine microbiota. Climate change and the increasing pressure of human activity on the ocean, including pollution, acidification and invasive species, are among the main challenges for the future. Projects producing and using basic research data in an integrative and transdisciplinary way offer multiple perspectives in understanding changes in ecosystem functioning and, consequently, are essential to support public policies for the conservation and sustainable use of marine biodiversity at different scales. UNESCO's Decade of Ocean (starting 2021) is a window of opportunity to strengthen marine research, to promote national and international collaboration, to build up networks involving the public and private sector, but particularly to draw society's attention to the importance of knowing marine environments and using ocean resources in a sustainable way. The advancement of ocean literacy is one of the main legacies for future generations promoted by integrated research programs such as BIOTA-FAPESP.

Resumo Neste estudo levantamos e analisamos 300 projetos relacionados à biodiversidade marinha financiados pela FAPESP entre 1972 a 2021, dos quais 46 foram realizados no Programa BIOTA. De um projeto único na década de 1970, o número foi aumentando gradativamente até 2009, quando o BIOTA promoveu uma chamada sobre biodiversidade marinha, o que impulsionou o número de projetos financiados nos anos seguintes. A abrangência geográfica dos projetos se expandiu ao longo dos anos e, a partir de estudos baseados no litoral do Estado de São Paulo, o foco gradualmente se deslocou para áreas mais amplas da costa brasileira, depois para outras áreas do Atlântico, e acabou se tornando global. A maioria dos projetos se concentrou em organismos costeiros, bentônicos, em substrato consolidado. Em termos de táxons, seis grupos responderam por cerca de 60% dos projetos (Crustacea, Actinopterygii, Mollusca, Chondrichthyes, Cnidaria e Rhodophyta), mas observa-se um aumento no número de grupos estudados ao longo das décadas. Os 300 projetos referem-se a um conjunto de 82 temas diferentes, dos quais os cinco principais são taxonomia, filogenia, comunidade, "ômicas" e poluição. As análises mostram um esforço de longa data em pesquisas de biodiversidade marinha, com abordagens atualizadas em relação ao escopo e métodos. A pesquisa em áreas estratégicas é discutida, incluindo os estudos sobre a microbiota marinha e em águas profundas. As mudanças climáticas e a crescente pressão da atividade humana sobre o oceano, incluindo poluição, acidificação e espécies invasoras, estão entre os principais desafios para o futuro. Projetos de produção e uso de dados de pesquisa básica de forma integrada e transdisciplinar oferecem múltiplas perspectivas de compreensão das mudanças no funcionamento dos ecossistemas e, consequentemente, são essenciais para subsidiar políticas públicas de conservação e uso sustentável da biodiversidade marinha em diferentes escalas. A Década do Oceano da UNESCO (a partir de 2021) é uma janela de oportunidade para fortalecer a pesquisa marinha, promover a colaboração nacional e internacional, construir redes envolvendo os setores público e privado, mas principalmente para chamar a atenção da sociedade para a importância de conhecer os ambientes marinhos e utilizar os recursos oceânicos de forma sustentável. O avanço da "alfabetização" oceânica é um dos principais legados para as gerações futuras promovidos por programas integrados de pesquisa como o BIOTA-FAPESP.

Xylella and Xanthomonas Mobil'omics.

The gamma-proteobacterium Xanthomonadales groups two closely related genera of plant pathogens, Xanthomonas and Xylella. Whole genome sequencing and comparative analyses disclosed a high degree of identity and co-linearity of the chromosome backbone between species and strains. Differences observed are usually clustered into genomic islands, most of which are delimited by genetic mobile elements. Focus is given in this paper to describe which groups of mobile elements are found and what is the relative contribution of these elements to Xanthomonas and Xylella genomes. Insertion sequence (IS) elements have invaded the Xanthomonas genome several times, whereas Xylella is rich in phage-related regions. Also, different plasmids are found inhabiting the bacterial cells studied here. Altogether, these results suggest that the integrative elements such as phages and transposable elements as well as the episomal plasmids are important drivers of the genome evolution of this important group of plant pathogens.

Unveiling privacy: advances in microtomography of coralline algae.

Marine calcareous algae are widespread in oceans of the world and known for their calcified cell walls and the generation of rhodolith beds that turn sandy bottoms into a complex structured ecosystem with high biodiversity. Rhodoliths are unattached, branching, crustose benthic marine red algae; they provide habitat for a rich variety of marine invertebrates. The resultant excavation is relevant to sediment production, while is common that the fragments or the whole specimens result in vast fossil deposits formed by rich material that can be "mined" for biological and geological data. Accordingly, microtomography (µCT) may enable a detailed investigation of biological and geological signatures preserved within the rhodolith structure in a non-destructive approach that is especially relevant when analyzing herbaria collections or rare samples. Therefore, we prepared coralline algae samples and submitted them to a range of capabilities provided by the SkyScan1176 micro-CT scanner, including reconstruction, virtual slicing, and pinpointing biological and geological signatures. To this end, polychaetes and mollusk shells, or their excavations, coral nucleation, sediment deposits and conceptacles were all observed. Although a similar technique has been applied previously to samples of living rhodoliths in Brazil, we show, for the first time, its successful application to fossil rhodoliths. We also provide a detailed working protocol and discuss the advantages and limitations of the microtomography within the rhodoliths.

The genome sequence of the gram-positive sugarcane pathogen Leifsonia xyli subsp. xyli.

The genome sequence of Leifsonia xyli subsp. xyli, which causes ratoon stunting disease and affects sugarcane worldwide, was determined. The single circular chromosome of Leifsonia xyli subsp. xyli CTCB07 was 2.6 Mb in length with a GC content of 68% and 2,044 predicted open reading frames. The analysis also revealed 307 predicted pseudogenes, which is more than any bacterial plant pathogen sequenced to date. Many of these pseudogenes, if functional, would likely be involved in the degradation of plant heteropolysaccharides, uptake of free sugars, and synthesis of amino acids. Although L. xyli subsp. xyli has only been identified colonizing the xylem vessels of sugarcane, the numbers of predicted regulatory genes and sugar transporters are similar to those in free-living organisms. Some of the predicted pathogenicity genes appear to have been acquired by lateral transfer and include genes for cellulase, pectinase, wilt-inducing protein, lysozyme, and desaturase. The presence of the latter may contribute to stunting, since it is likely involved in the synthesis of abscisic acid, a hormone that arrests growth. Our findings are consistent with the nutritionally fastidious behavior exhibited by L. xyli subsp. xyli and suggest an ongoing adaptation to the restricted ecological niche it inhabits.

In vitro assay and light regulation of nitrate reductase in red alga Gracilaria chilensis.

Nitrate reductase (NR) is the first enzyme in the nitrogen assimilation pathway. The in vitro NR activity of Gracilaria chilensis was assayed under different conditions to reveal its stability and biochemical characteristics, and an optimized in vitro assay is described. Maximal NR activities were observed at pH 8.0 and 15 degrees C. The apparent Km value for NADH was 8 microM and for nitrate 680 microM. Crude extracts of G. chilensis stored at 4 degrees C showed a 50% decrease of NR activity after 24 h. The highest NR activity value (253.20+/-2.60 x 10(-3) U g(-1)) was obtained when 100% von Stosch medium (500 microM NO3-) was added before extraction of apical parts. Algae under light:dark cycles of 12:12h exhibited circadian fluctuation of NR activity and photosynthesis with more than 2 times higher levels in the light phase. No evidence of endogenous diel rhythm controlling NR activity or photosynthesis was observed. Light pulses lasting 10 or 60 min during the darkness increased the NR activity by 30% and 45%, respectively. The results indicate that NR and photosynthesis are regulated mainly by light and not by a biological clock.

A NEW LOOK AT AN ANCIENT ORDER: GENERIC REVISION OF THE BANGIALES (RHODOPHYTA)(1).

The red algal order Bangiales has been revised as a result of detailed regional studies and the development of expert local knowledge of Bangiales floras, followed by collaborative global analyses based on wide taxon sampling and molecular analyses. Combined analyses of the nuclear SSU rRNA gene and the plastid RUBISCO LSU (rbcL) gene for 157 Bangiales taxa have been conducted. Fifteen genera of Bangiales, seven filamentous and eight foliose, are recognized. This classification includes five newly described and two resurrected genera. This revision constitutes a major change in understanding relationships and evolution in this order. The genus Porphyra is now restricted to five described species and a number of undescribed species. Other foliose taxa previously placed in Porphyra are now recognized to belong to the genera Boreophyllum gen. nov., Clymene gen. nov., Fuscifolium gen. nov., Lysithea gen. nov., Miuraea gen. nov., Pyropia, and Wildemania. Four of the seven filamentous genera recognized in our analyses already have generic names (Bangia, Dione, Minerva, and Pseudobangia), and are all currently monotypic. The unnamed filamentous genera are clearly composed of multiple species, and few of these species have names. Further research is required: the genus to which the marine taxon Bangia fuscopurpurea belongs is not known, and there are also a large number of species previously described as Porphyra for which nuclear SSU ribosomal RNA (nrSSU) or rbcL sequence data should be obtained so that they can be assigned to the appropriate genus.

Estudo sobre a sensibilidade dos caprinos à toxidez de crotalárias tóxicas para bovinos visando a sua utilização na profilaxia / Study about the sensitivity of goats to the toxicity of crotalarias poisonous for cattle aiming their use for prophylactic measures

A intoxicação por plantas é sabidamente uma das principais causas de morte de bovinos adultos no Brasil. Uma vez que pouco pode ser feito no que diz respeito ao tratamento destas intoxicações, os esforços devem se concentrar na adoção de medidas profiláticas alternativas, como a utilização de uma espécie menos sensível no local de ocorrência de determinadas plantas. Objetivou-se com esse trabalho verificar a sensibilidade de caprinos às crotalárias (Crotalaria spectabilis e Crotalaria juncea) para que estes possam ser utilizados na profilaxia da intoxicação por essas plantas em bovinos. Sementes de C. spectabilis e C. juncea foram fornecidas em doses únicas ou diárias a oito caprinos adultos jovens, de ambos os sexos, divididos aleatoriamente em dois grupos de quatro animais (um grupo para cada planta)...

Plant poisoning is known to be one of the most important causes of death of adult cattle in Brazil. As there are not many possibilities for treatment, all efforts must focus on application of alternative prophylactic measures, among them the use of a less sensitive animal species in areas of the occurrence of certain plants...

GRACILARIOPSIS MCLACHLANII SP. NOV. AND GRACILARIOPSIS PERSICA SP. NOV. OF THE GRACILARIACEAE (GRACILARIALES, RHODOPHYCEAE) FROM THE INDIAN OCEAN(1).

Two new species of Gracilariopsis from the Indian Ocean are proposed-Gracilariopsis (Gp.) mclachlanii Buriyo, Bellorin et M. C. Oliveira sp. nov. from Tanzania and Gracilariopsis persica Bellorin, Sohrabipour et E. C. Oliveira sp. nov. from Iran-based on morphology and DNA sequence data (rbcL gene and SSU rDNA). Both species fit the typical features of Gracilariopsis: axes cylindrical throughout, freely and loosely ramified up to four orders, with an abrupt transition in cell size from medulla to cortex, cystocarps lacking tubular nutritive cells and superficial spermatangia. Nucleotide sequence comparisons of rbcL and SSU rDNA placed both species into the Gracilariopsis clade as distinct species from all the accepted species for this genus, forming a deeply divergent lineage together with some species from the Pacific. The new species are very difficult to distinguish on morphological grounds from other species of Gracilariopsis, stressing the importance of homologous molecular marker comparisons for the species recognition in this character-poor genus.

Morphological and molecular studies on the Brazilian native red seaweed Laurencia oliveirana (Rhodomelaceae, Ceramiales)

Morphological and molecular studies were carried out on Laurencia oliveirana from the type locality (Arraial do Cabo, Rio de Janeiro, Brazil). This species is easily recognized by its small size, sub-erect habit forming intricate cushion-like tufts and unilateral pectinate branching. The species displays all the typical characters of the genus Laurencia, such as the production of the first pericentral cell underneath the basal cell of the trichoblast, tetrasporangia produced from particular pericentral cells, with the third and fourth pericentral cells becoming fertile, without production of additional pericentral cells, spermatangial branches produced from one of two laterals on the suprabasal cell of trichoblasts, and procarp-bearing segment with five pericentral cells. Details of tetrasporangial plants and development of procarp and male plants are described for the first time for the species. The phylogenetic position of L. oliveirana was inferred by analysis of the chloroplast-encoded rbcL gene sequences from 57 taxa. In all phylogenetic analyses, L. oliveirana grouped with L. caraibica, L. caduciramulosa, L. venusta and L. natalensis, forming a monophyletic clade within the Laurencia sensu stricto. The genetic divergence between L. oliveirana and the molecularly closest species, L. caraiba collected in Brazil, was 2.3%.