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Complex small supernumerary marker chromosomes (sSMCs) consist of chromosomal material derived from two or more different chromosomal regions and constitute one of the smallest subsets of sSMC. Most of complex sSMCs are represented by a der(22)t(11;22) in Emanuel syndrome. As far as we know, only one recent report has described sSMCs involving simultaneously X and Y chromosomes in Turner Syndrome. We report two patients, a female and a male, both with a complex sSMC derived from X and Y chromosomes in mosaic with a 45,X cell line. In both patients, the marker chromosomes were early replicating and the XIST gene was absent. FISH and PCR confirmed the presence of Yp loci (TSPY, AMGY, SRY, DYZ3), and negative for DYZ1. The DAZ4 sequence was present only in patient 1.Our findings suggested that complex sSMC involving X and Y chromosome could be a kind of sSMC of the gonosomes.

Congenital Zika Syndrome (CZS) is a set of birth defects caused by Zika virus (ZIKV) infection during pregnancy. Microcephaly is its main feature, but other brain abnormalities are found in CZS patients, such as ventriculomegaly, brain calcifications, and dysgenesis of the corpus callosum. Many studies have focused on microcephaly, but it remains unknown how ZIKV infection leads to callosal malformation. To tackle this issue, we infected mouse embryos in utero with a Brazilian ZIKV isolate and found that they are born with a reduction in callosal area and density of callosal neurons. ZIKV infection also causes a density reduction of PH3+ cells, intermediate progenitor cells and SATB2+ neurons. Moreover, axonal tracing revealed that callosal axons are reduced and misrouted. Also, ZIKV infected cultures show a reduction of callosal axon length. GFAP labelling showed that in utero infection compromises glial cells responsible for midline axon guidance. The RNA-Seq data from infected brains identified downregulation of axon guidance and axonogenesis related genes. In sum, we showed that ZIKV infection impairs critical steps of corpus callosum formation by disrupting not only neurogenesis but also axon guidance and growth across the midline.Summary StatementZika virus infection during development impairs the formation of corpus callosum by disturbing axon guidance and growth of callosal neurons.

Genus Arachis comprises 82 species distributed into nine taxonomic sections. Most Arachis species are wild and those from Arachis section have been evaluated for many traits, since they can be used in peanut breeding. Most of the remaining species have been neglected and understudied. Recently, resveratrol content and expression of a resveratrol synthase gene were analyzed in wild Arachis species. Our aim was to expand the knowledge about resveratrol in Arachis, analyzing species from five sections and evaluating the expression of a resveratrol synthase (RS) gene responsive to ultraviolet light (UV) along the time. In a first experiment, the resveratrol content after UV induction was analyzed on detached leaves of 12 species from five sections. Variation was observed among species and accessions of the same species. The highest contents were found in A. lignosa (843.9 μg/g) and A. triseminata (745.4 μg/g). In a second experiment, RS expression and resveratrol content in four species and one synthetic amphidiploid were analyzed at 0, 7, 15 and 24 h pos induction (hpi) with UV. In most genotypes, the highest RS expression level was at 0 hpi, whereas the highest resveratrol content was at 15 hpi. Our results suggested that resveratrol is ubiquitously present in the genus Arachis with different capacities of synthesis among species and accessions in response to ultraviolet treatment. Presence of resveratrol in wild Arachis species adds new value to these genetic resources.

Herbivory is one of the major biotic stress factors that affect the establishment of plants. However, the main factors that drive herbivory in seedlings of Amazonian tree species are still not well understood. Here we investigated whether contrasting levels of irradiance influence herbivory according to different herbivory indicators and which leaf traits are most related to interspecific variation in herbivory under contrasting irradiance conditions. We measured the leaf area lost as a result of insect herbivory in five tree species planted in a silvicultural system of secondary forest enrichment according to two indicators, herbivore damage (accumulated since plant germination) and herbivory rate (measured over time), and two irradiance conditions, understorey PPFD 2.6 mol·m-2 ·day-1 ) and gap PPFD 33.1 mol·m-2 ·day-1 . Furthermore, we related the interspecific variation in herbivory to a set of leaf traits: SLA, RWC, sclerophylly, phenolic compound content, tannins, condensed tannins and non-structural carbohydrates. Herbivore damage was significantly affected by light availability and species, with the highest percentage variation observed in the Meliaceae (Carapa guianensis and Swietenia macrophylla). For the herbivory rate, only the interspecific variation was significant, with Bertholletia excelsa having the lowest rates. Chemical characteristics (phenolic compounds and tannins) were most related to herbivory rates, as well as highly influenced by light conditions. Non-structural carbohydrates (starch and sucrose) were also related to the interspecific variation in herbivory. The phenolic compounds and starch, as affected by light quantity, are species dependent. Thus, the selective pressure on herbivores may be driven by species-dependent responses to light conditions.

Plant parasitic root-knot nematodes (RKN) have evolved sophisticated strategies for exploiting plants, and they cause devastating yield losses in many susceptible crops. The RKN Meloidogyne arenaria is the most predominant pathogenic nematode affecting cultivated peanut (Arachis hypogaea L.). Resistance against the M. arenaria has been identified in the wild peanut relative, Arachis stenosperma. Transcriptome studies on A. stenosperma–M. arenaria interaction revealed candidate genes potentially involved in the first stages of this resistance response, including a temperature-induced lipocalin (TIL) protein. Plant TILs have a protective role against harmful molecules produced in response to a number of stresses. In this study, the characterization of the RKN-responsive TIL from A. stenosperma (AsTIL) provides new insights into the role of plant lipocalins in nematode resistance. The AsTIL gene has a 2-exon/1-intron structure and encodes a 21.4 kDa polypeptide. It contains three structurally conserved regions, which are a signature for plant lipocalins. Overexpression of AsTIL in transgenic hairy roots from a susceptible peanut cultivar led to the reduction of M. arenaria galls and egg masses 60 days after inoculation. This is the first report of a possible involvement of plant lipocalins in RKN resistance. It also reveals a promising candidate gene for peanut breeding to produce cultivars resistant to M. arenaria.

Peanut wild relatives (Arachis spp.) have high genetic diversity and are important sources of resistance to biotic and abiotic stresses. In this study, proteins were analyzed in root tissues of A. duranensis submitted to a progressive water deficit in soil and the differential abundance was compared to transcript expression profiles obtained by RNA-seq and qRT-PCR. Using a 2-DE approach, a total of 31 proteins were identified, most of which were associated with stress response and drought perception. These comprised a chitinase-2 (unique to stressed condition), an MLP-like protein, a glycine-rich protein DOT1-like, a maturase K and heat shock-related proteins (HSP70 - an isoform unique to the control, and HSP17.3). Other proteins unique to the control condition comprised a transcription initiation factor IIF subunit alpha-like protein, a SRPBCC ligand-binding domain superfamily protein, an Adenine phosphoribosyl transferase, a Leo1-like protein, a Cobalamine-independent methionine synthase and a Transmembrane emp24 domain-containing protein p24delta9-like. Correlation of mRNA expression and corresponding protein abundance was observed for 15 of the identified proteins, with genes encoding the majority of proteins (14) negatively regulated in stressed roots. Proteins identified in this study offer potential for the genetic improvement of cultivated peanut for drought tolerance. SIGNIFICANCE: The comparison of protein abundance and corresponding transcript expression levels (RNA-seq and qRT-PCR) revealed that 15 of the identified proteins showed similar expression behavior, with the majority (14 proteins) negatively regulated in stressed roots. Chitinase-2 (Cht2) was the only protein with an upregulation behavior in all approaches. These proteins appear to play an important role in drought tolerance in A. duranensis and may be further explored in peanut genetic breeding programs.

The use of drought-tolerant cultivars is a promising strategy to minimize the effects of water deficits on upland rice productivity. Therefore, it is important to establish an adequate methodology to evaluate the performance of upland rice cultivars under water deficit. In this study, one evaluated the effects of water deficit on morphological and physiological traits of two upland rice cultivars, in a greenhouse, in soil columns, using a phenotyping platform named SITIS. The experiment occurred in a complete randomized block design, in a split plot scheme with four replications. The main plots consisted of a well-irrigated treatment with daily replacement of 100% evapotranspired water and four water restriction regimes (daily replacement of 20%, 40%, 60% and 80% evapotranspired water at the well-irrigated treatment) and, the subplots, of the Douradão and BRS Soberana cultivars, which are tolerant and sensitive to water stress, respectively. It is feasible to use the SITIS platform and the proposed protocol (water regimes) to evaluate upland rice cultivars for tolerance to water stress. Under water deficit, the Douradão cultivar restricted its transpiration and showed harvest index, grain yield and water use efficiency higher than the BRS Soberana cultivar. Made available in DSpace on 2019-01-25T23:53:19Z (GMT). No. of bitstreams: 1 CNPAF2018rbai.pdf: 762741 bytes, checksum: da3a1364685879c6cc44ea16526140ca (MD5) Previous issue date: 2019-01-24

Although it is well-established that severe poisoning by organophosphorus (OP) compounds strongly affects the cardiorespiratory system, the effects of sub-lethal exposure to these compounds on the neural control of cardiovascular function are poorly explored. The aim of this study was to evaluate the effects of acute sub-lethal exposure to chlorpyrifos (CPF), a commonly used OP insecticide, on three basic reflex mechanisms involved in blood pressure regulation, the peripheral chemoreflex, the baroreflex and the Bezold-Jarisch reflex. Adult male Wistar rats were injected intraperitoneally with a single dose of CPF (30 mg/kg) or saline (0.9%). 24 h after injections, cardiovascular reflexes were tested in awake rats. Potassium cyanide (KCN) and phenylbiguanide (PBG) were injected intravenously to activate the chemoreflex and the Bezold-Jarisch reflex, respectively. The baroreflex was activated by phenylephrine and sodium nitroprusside infusions. Blood samples were taken for measurements of butyrylcholinesterase (BChE) activity while acetylcholinesterase (AChE) activity was measured in brainstem samples. Animals treated with CPF presented signs of intoxication such as ataxia, tremor, lacrimation, salivation, tetany, urination and defecation. The hypertensive and the bradycardic responses of the chemoreflex as well as the hypotensive and bradycardic responses of the Bezold-Jarisch reflex were attenuated in CPF treated animals (P < 0.05). Concerning the baroreflex responses, CPF treatment reduced the bradycardia plateau, the range and the gain of the reflex (P < 0.05). Plasma BChE and brainstem AChE were both reduced significantly after CPF treatment (P < 0.05). Our results showed that acute sub-lethal exposure to CPF impairs the cardiovascular responses of homeostatic and defensive cardiovascular reflexes. These effects are associated with a marked inhibition of plasma BChE and brainstem AChE.

Peanut,Arachishypogaea(Linnaeus, 1753) is an allotetraploid cultivated plant with two subgenomes derived from the hybridization between two diploid wild species,A.duranensis(Krapovickas & W. C. Gregory, 1994) andA.ipaensis(Krapovickas & W. C. Gregory, 1994), followed by spontaneous chromosomal duplication. To understand genome changes following polyploidy, the chromosomes ofA.hypogaea, IpaDur1, an induced allotetraploid (A.ipaensis×A.duranensis)4xand the diploid progenitor species were cytogenetically compared. The karyotypes of the allotetraploids share the number and general morphology of chromosomes; DAPI+bands pattern and number of 5S rDNA loci. However, one 5S rDNA locus presents a heteromorphic FISH signal in both allotetraploids, relative to corresponding progenitor. Whilst forA.hypogaeathe number of 45S rDNA loci was equivalent to the sum of those present in the diploid species, in IpaDur1, two loci have not been detected. Overall distribution of repetitive DNA sequences was similar in both allotetraploids, althoughA.hypogaeahad additional CMA3+bands and few slight differences in the LTR-retrotransposons distribution compared to IpaDur1. GISH showed that the chromosomes of both allotetraploids had preferential hybridization to their corresponding diploid genomes. Nevertheless, at least one pair of IpaDur1 chromosomes had a clear mosaic hybridization pattern indicating recombination between the subgenomes, clear evidence that the genome of IpaDur1 shows some instability comparing to the genome ofA.hypogaeathat shows no mosaic of subgenomes, although both allotetraploids derive from the same progenitor species. For some reasons, the chromosome structure ofA.hypogaeais inherently more stable, or, it has been at least, partially stabilized through genetic changes and selection.

Peanut is a crop of the Kayabi tribe, inhabiting the Xingu Indigenous Park, Brazil. Morphological analysis of Xingu accessions showed variation exceeding that described for cultivated peanuts. This raised questions as to the origin of the Xingu accessions: are they derived from different species, or is their diversity a result of different evolutionary and selection processes? To answer these questions, cytogenetic and genotyping analyses were conducted. The karyotypes of Xingu accessions analyzed are very similar to each other, to an A. hypogaea subsp. fastigiata accession and to the wild allotetraploid A. monticola. The accessions share the number and general morphology of the chromosomes; DAPI+ bands; 5S and 45S rDNA loci distribution and a high genomic affinity with A. duranensis and A. ipaënsis genomic probes. However, the number of CMA3 + bands differs from those determined for A. hypogaea and A. monticola, which are also different from each other. SNP genotyping grouped all Arachis allotetraploids into four taxonomic groups: Xingu accessions were closer to A. monticola and A. hypogaea subsp. hypogaea. Our data suggests that the morphological diversity within these accessions is not associated with a different origin and can be attributed to morphological plasticity and different selection by the Indian tribes.

Background: Osteosarcoma (OS) is the most-common primary bone tumor among children and adolescents, presenting a high risk of metastasis. Despite the OS incidence peak in the second decade of life, it has been shown that a significant number of children are diagnosed with less than 10 years of age. The investigation of possible biological and clinical particularities in this group of patients can contribute to a better understanding of OS tumorigenesis and to development of a more personalized therapeutic and clinical management. The aim of this study is to identify genetic alterations in children diagnosed with OS in an unexpected age using next-generation sequencing (NGS) strategy. Methods: Thirty OS samples from 30 patients who were under the age of 10 at the time of diagnosis, treated at the Pediatric Oncology Institute GRAACC/UNIFESP, were submitted to NGS using a specific panel for childhood and adolescent neoplasm named OCCRA (Oncomine Childhood Cancer Research Assay), from Thermo Fisher. Clinical variables have also been collected. Results: Of the 30 samples analyzed 16 (53.3%) presented some genetic alteration. The most commonly genetic alteration identified was CNVs in the MYC gene, a proto-oncogene that has already been associated with worse prognosis and response to treatment in OS cases. All 6 patients with MYC amplification presented bad response to treatment. ARID1A tumor suppressor gene was the second-most commonly mutated, with 4 patients presenting loss-of-function InDels, not reported in COSMIC database. We also identified 3 patients with CDK4 CNVs classified as gain-of-function alterations and 3 patients with loss-of-function TP53 alterations, 2 SNVs and 1 InDel. Two of the identified variants in TP53 have not yet been reported in COSMIC database. Of the 3 patients harboring TP53 variants, one has Li-Fraumeni syndrome confirmed. Conclusions: The genetic profiling based on a specific panel for children and adolescent cancer, including non-reported variants identification in patients diagnosed with OS in an unexpected age, is essential for a more accurate tumor profiling and the identification of risk groups. The investigation of mutations with clinical relevance can also contribute to a more precise therapeutic management. NGS strategy can lead into a better understanding about these questions. Funding Source: São Paulo Research Foundation (FAPESP) - n° 2019/13056-0 Citation Format: Giovanna M. Guimarães, Francine Tesser Gamba, Antônio Sérgio Petrilli, Maria Teresa S. Alves, Reinaldo J. Garcia-Filho, Renato de Oliveira, Sílvia Regina C. Toledo. Osteosarcoma under the age of 10: using next-generation sequencing panel for tumor profile investigation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2248.

Premise of the study Several species of Arachis have been cultivated for their edible seeds, historically and to the present day. The diploid species that have a history of cultivation show relatively small signatures of domestication. In contrast, the tetraploid species A. hypogaea evolved into highly domesticated forms and became a major world crop, the cultivated peanut. It seems likely that allotetraploidization (hybridity and/or tetraploidization) in some way enhanced attractiveness for cultivation. Here we investigate this using six different hybridization and tetraploidization events, from distinct Arachis diploid species, including one event derived from the same wild species that originated peanut. Methods Twenty-six anatomical, morphological, and physiological traits were examined in the induced allotetraploid plants and compared with their wild diploid parents. Key results Nineteen traits were transgressive (showed strong response to hybridization and chromosome duplication): allotetraploids had larger leaves, stomata and epidermal cells than did their diploid parents. In addition, allotetraploids produced more photosynthetic pigments. These traits have the same trend across the different hybrid combinations, suggesting that the changes are more likely due to ploidy rather than hybridity. In contrast, seed dimensions and seed mass did not significantly change in response to hybridization or tetraploidization. Conclusions We suggest that the original allotetraploid that gave rise to cultivated peanut may have been attractive because of an increase in plant size, different transpiration characteristics, higher photosynthetic capacity, or other characteristics, but contrary to accepted knowledge, increased seed size was unlikely to have been important in the initial domestication.

The development of neuropathic pain after peripheral nerve injury involves neuroimmune–glial interactions in the spinal cord. However, whether the development of neuropathic pain depends on the infiltration of peripheral immune cells, such as monocytes, into the spinal cord parenchyma after peripheral nerve damage remains unclear. Here, we used a combination of different techniques such as transgenic reporter mouse (Cx3cr1GFP/+ and Ccr2RFP/+ mice), bone marrow chimeric mice, and parabiosis to investigate this issue in spared nerve injury (SNI) model. Herein, we provided robust evidence that, although microglial cells are activated/proliferate at the dorsal horn of the spinal cord after SNI, peripheral hematopoietic cells (including monocytes) are not able to infiltrate into the spinal cord parenchyma. Furthermore, there was no evidence of CCR2 expression in intrinsic cells of the spinal cord. However, microglial cells activation/proliferation in the spinal cord and mechanical allodynia after SNI were reduced in Ccr2-deficient mice. These results suggest that blood-circulating leukocytes cells are not able to infiltrate the spinal cord parenchyma after distal peripheral nerve injury. Nevertheless, they indicate that CCR2-expressing cells might be indirectly regulating microglia activation/proliferation in the spinal cord after SNI. In conclusion, our study supports that CCR2 inhibition could be explored as an interventional approach to reduce microglia activation and consequently neuropathic pain development after peripheral nerve injury.

In 2008 and 2009, vine decline symptoms were observed in three watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) fields located in the municipalities of Mossoró (Rio Grande do Norte State) and Quixeré (Ceará State) in northeastern Brazil. Symptoms included yellowing of crown leaves just prior to harvest and collapse of many of the vines. Mean maximum daily temperatures for the first and second half of the season were 28.6 and 25.1°C, respectively. Affected plants exhibited necrotic root systems and lacked most of the secondary and tertiary feeder roots. Numerous perithecia on the roots contained asci and ascospores characteristics of Monosporascus cannonballus Pollack & Uecker (1,2). Small pieces of primary and secondary roots were surface disinfected and plated onto potato dextrose agar (PDA) medium with 0.5 g liter–1 of streptomycin sulfate and incubated for 7 days at 25°C in the dark. Hyphal tips from all colonies were transferred to PDA and further incubated for 30 to 40 days at 25°C in the dark for subsequent growth and sporulation. Isolations consistently yielded colonies of white mycelium, which became dark grayish after 10 to 15 days, and perithecia with one-spored asci. The internal transcribed spacer regions of ribosomal DNA of isolates 18-5 and 19-1 were sequenced (GenBank Accession Nos. GQ891544 and GQ891545). These sequences were identical to sequences of M. cannonballus (GenBank Accession Nos. AM167936 and AM167937). Pathogenicity of these two isolates was confirmed on watermelon cv. Crimson Sweet in a greenhouse maintained at 25 to 30°C. Inoculum was produced in a sand-oat hulls (Avena sativa) medium (0.5 liter of sand, 46 g of ground oat hulls, and 37.5 ml of distilled water) and incubated at 25°C for 1 month. CFU were quantified by serial dilution using 1% hydroxyethyl cellulose. A sterilized mixture of equal portions (vol/vol) of sand and peat moss was used to fill 17-cm-diameter plastic pots and inoculum was added to produce an inoculum concentration of 20 CFU g–1. Five watermelon seeds planted in each pot were later thinned to one seedling per pot. There were five replicated pots for each treatment with an equal number of noninfested pots. Plants were evaluated for disease 45 days after sowing. All isolates of M. cannonballus were highly aggressive and caused severe root necrosis compared with the noninoculated controls. M. cannonballus was reisolated from symptomatic plants, confirming Koch's postulates. In 2004, M. cannonballus was reported in the same Brazilian cucurbit-growing areas causing root rot and vine decline of muskmelon (Cucumis melo L.) (3), but to our knowledge, this is the first report of M. cannonballus on watermelon in Brazil. References: (1) R. D. Martyn and M. E. Miller. Plant Dis. 80:716, 1996. (2) F. G. Pollack and F. A. Uecker. Mycologia 66:346, 1974. (3) R. Sales Jr. et al. Plant Dis. 88:84, 2004.

The central nervous system is a highly complex network composed of various cell types, each one with different subpopulations. Each cell type has distinct roles for the functional operation of circuits, and ultimately, for brain physiology in general. Since the absolute number of each cell type is considered a proxy of its functional complexity, one approach to better understand how the brain works is to unravel its absolute cellularity and the quantitative relations between cell populations; in other words, how one population of cells is quantitatively structured, in relation to another. Oligodendrocytes are one of these cell types - mainly, they provide electric insulation to axons, optimizing action potential conduction. Their function has recently been revisited and their role extended, one example being their capability of providing trophic support to long axons. To determine the absolute cellularity of oligodendroglia, we have developed a protocol of oligodendrocyte quantification using the isotropic fractionator with a pan-marker for this cell type. We report a detailed assessment of specificity and universality of the oligodendrocyte transcription factor 2 (Olig2), through systematic confocal analyses of the C57BL/6 mouse brain. In addition, we have determined the absolute number (17.4 million) and proportion (about 20%) of this cell type in the brain (and in different brain regions), and tested if this population, at the intraspecific level, scales with the number of neurons in an allometric-based approach. Considering these numbers, oligodendrocytes proved to be the most numerous of glial cells in the mouse brain.

Rust is a major pathogen of the peanut crop. Development and adoption of rust-resistant cultivars is the most cost efficient and effective way to control the spread of the disease and reduce yield losses. Some cultivated peanut germplasm accessions have a degree of resistance, but the secondary gene pool is a source of much stronger resistance alleles. Wild species, however, have undesirable agronomic traits that are a disincentive to their use in breeding. The identification of genomic regions that harbor disease resistance in wild species is the first step in the implementation of marker-assisted selection that can speed the introgression of wild disease resistances and the elimination of linkage drag. In this work, we identify genome regions that control different components of rust resistance in a recombinant inbred line population developed from a cross between two Arachis species, the susceptible most probable B genome ancestor of cultivated peanut, Arachis ipaënsis, and an accession of its closest relative, Arachis magna, which is resistant to rust. Quantitative trait loci for several components of resistance were placed in the same position on linkage group B08. Single-nucleotide polymorphism Kompetitive allele-specific polymerase chain reaction markers for rust resistance region were designed and validated for marker function in both diploid and tetraploid contexts.

This study aimed to evaluate the cultivation of forage sorghum subjected to different leaching fractions with saline effluent from fish-farming under semiarid conditions. The experiment was set in a randomized block design, with four blocks, in split plots, composed of four leaching fractions (0; 5; 10 and 15%) and three forage sorghum varieties ('Volumax', 'F305' and 'Sudão'). Irrigation was performed using saline effluent from fish farming with electrical conductivity of 2.5 dS m-1. The analyzed variables were: plant height; stem diameter; width, length and number of leaves; fresh and dry matter yield, and relative contents of potassium and sodium in the shoots. Forage sorghum under saline effluent irrigation and leaching fraction of 15% shows a yield increase of 25%, in comparison to sorghum without the leaching fraction. The variety 'Volumax' was more sensitive to salinity than the others, since it showed lower shoot growth and low values of leaf area, fresh matter and dry matter. RESUMO O presente trabalho objetivou avaliar o cultivo de variedades de sorgo forrageiro submetido a diferentes frações de lixiviação com efluente salino da piscicultura em condições semiáridas. Foi adotado o delineamento experimental em blocos ao acaso, com quatro blocos, em parcelas subdivididas compostas por quatro frações de lixiviação (0; 5; 10 e 15%) nas parcelas e três variedades de sorgo forrageiro (Volumax, F305 e Sudão) nas subparcelas. A irrigação foi realizada com efluente salino da piscicultura (CE de 2,5 dS m-1). As variáveis avaliadas foram altura da planta; diâmetro do colmo; largura, comprimento e número de folhas; produtividade de massa fresca e seca, além de teores de sódio e potássio na parte aérea. Observou-se que o sorgo forrageiro irrigado com efluente salino, com fração de lixiviação de 15% apresentou um aumento percentual de até 25% na produtividade quando comparada à do sorgo sem fração de lixiviação. A variedade Volumax mostrou-se mais sensível à salinidade que as demais em virtude de apresentar o menor crescimento da parte aérea e baixos valores de área foliar e matéria fresca e seca.

The Anopheles albitasis complex includes 6 species, and 3 are considered as malaria vectors in Brazil. Twenty-five polymorphic microsatellite DNA loci were isolated and characterized in 24-36 individuals from the neighborhood of Puraquequara, Manaus, Amazonas State, Brazil. The number of estimated alleles ranged from 2 to 10, the observed heterozygosity ranged from 0.182 to 0.897, and the expected heterozygosity ranged from 0.260 to 0.854. Eleven loci showed significant deviation from Hardy-Weinberg equilibrium. Eleven loci were cross-amplified successfully in 5 Anopheles species. These microsatellite loci will be useful in studies investigating population structure and evolutionary genetics in A. albitarsis sensu lato and other A. albitarsis complex species.

PREMISE OF THE STUDY The genetic bottleneck of polyploid formation can be mitigated by multiple origins, gene flow, and recombination among different lineages. In crop plants with limited origins, efforts to increase genetic diversity have limitations. Here we used lineage recombination to increase genetic diversity in peanut, an allotetraploid likely of single origin, by crossing with a novel allopolyploid genotype and selecting improved lines. METHODS Single backcross progeny from cultivated peanut × wild species-derived allotetraploid cross were studied over successive generations. Using genetic assumptions that encompass segmental allotetraploidy, we used single nucleotide polymorphisms and whole-genome sequence data to infer genome structures. KEY RESULTS Selected lines, despite a high proportion of wild alleles, are agronomically adapted, productive, and with improved disease resistances. Wild alleles mostly substituted homologous segments of the peanut genome. Regions of dispersed wild alleles, characteristic of gene conversion, also occurred. However, wild chromosome segments sometimes replaced cultivated peanut's homeologous subgenome; A. ipaensis B sometimes replaced A. hypogaea A subgenome (~0.6%), and A. duranensis replaced A. hypogaea B subgenome segments (~2%). Furthermore, some subgenome regions historically lost in cultivated peanut were "recovered" by wild chromosome segments (effectively reversing the "polyploid ratchet"). These processes resulted in lines with new genome structure variations. CONCLUSIONS Genetic diversity was introduced by wild allele introgression, and by introducing new genome structure variations. These results highlight the special possibilities of segmental allotetraploidy and of using lineage recombination to increase genetic diversity in peanut, likely mirroring what occurs in natural segmental allopolyploids with multiple origins.

Made available in DSpace on 2018-12-11T17:37:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) In tropical conservation agricultural systems, crop yield is limited by soil acidity and root-growth inhibition, especially under intensive crop rotation. This study evaluated the effect of surface applications of lime and phosphogypsum in improving soil fertility and crop yield in a tropical region. Four treatments were evaluated: control (without soil amendment) and application phosphogypsum (2.1 + 2.1 + 2.1 Mg ha -1), lime (2.7 + 2.0 + 2.0 Mg ha -1), and a combination of lime and phosphogypsum at the given rates, applied in 2002, 2004 and 2010, respectively. We evaluated the soil chemical properties, root development, plant nutrition, yield components and grain yield of 10 crops over 4 years using five species: maize (Zea mays), crambe (Crambe abyssinica), cowpea (Vigna unguiculata), wheat (Triticum aestivum) and common bean (Phaseolus vulgaris). Our long-term results demonstrate the benefits of surface liming in alleviating subsoil acidity, reducing Al 3+ toxicity, improving availability of Ca 2+ and Mg 2+, and increasing accumulation of soil organic matter in all soil profiles at depths up to 0.60 m. For maize and crambe, adding phosphogypsum increased development of plants and reproductive structures, which increased grain yield. Phosphogypsum exhibited synergistic effects in association with lime for maize and common bean. Phosphogypsum did not have an effect on cowpea and wheat, whereas surface liming was essential to improve plant nutrition, grain yield and wheat grain quality. The combination of both soil amendments is an important tool to reduce the soil acidification process, resulting in the highest levels of Ca 2+ and Mg 2+ and the highest base-saturation values in the topsoil layers (0-0.20 m) over time. Our long-term results showed the viability of surface liming plus phosphogypsum for improving tropical soil fertility, which can reflect an increase in grain yield and contribute to the sustainability of agricultural systems under intensive land use in highly weathered areas. Special Academic Unit of Agricultural Sciences Federal University of Goiás (UFG) College of Agricultural Sciences Department of Crop Science São Paulo State University (FCA/UNESP), PO Box 237 College of Agricultural Sciences Department of Crop Science São Paulo State University (FCA/UNESP), PO Box 237

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder characterized by motor dysfunction, cognitive deficits, and psychiatric symptoms. The primary genetic cause is an expansion of cytosine adenine guanine (CAG) nucleotides of the huntingtin gene, which codes an important protein involved with neuronal signaling. The severity of HD correlates with the number of CAG repeats and individuals with longer expansions have an earlier onset and more severe symptoms. A microarray study conducted by our research group showed alteration in DNAH6 gene (encoding dynein axonemal heavy chain 6). DNAH6 belongs to dynein family, whose members are constituents of the microtubule-associated motor proteins and is downregulated in the striatum of a HD mouse model (knockin HdhQ111/Q111). In this manner, our goal was to confirm these downregulations in the mouse model and verify if the same alteration in the axonemal DNAH6 gene expression is observed in blood samples of HD patients. Blood samples were collected from 17 patients with clinical diagnosis of HD and 12 healthy individuals and RNA extracted for qPCR analysis. Microarray data were confirmed by qPCR in knockin HdhQ111/Q111, and DNAH6 was severely decreased in those mice, as compared to control mice (HdhQ20/Q20). Notably, decreased expression of DNAH6 gene was also observed in HD patients when compared to control group and negatively correlates with the CAG expansion. Although further studies are necessary to underlie the molecular mechanisms of dynein-htt interaction, this data highlights DNAH6 as a potential new blood marker for HD.

Background Peanut (Arachis hypogaea) production is largely affected by a variety of abiotic and biotic stresses, including the root-knot nematode (RKN) Meloidogyne arenaria that causes yield losses worldwide. Transcriptome studies of wild Arachis species, which harbor resistance to a number of pests and diseases, disclosed several candidate genes for M. arenaria resistance. Peanut is recalcitrant to genetic transformation, so the use of Agrobacterium rhizogenes-derived hairy roots emerged as an alternative for in-root functional characterization of these candidate genes. Results The present report describes an ex vitro methodology for hairy root induction in detached leaves based on the well-known ability of peanut to produce roots spontaneously from its petiole, which can be maintained for extended periods under high-humidity conditions. Thirty days after infection with the A. rhizogenes ‘K599’ strain, 90% of the detached leaves developed transgenic hairy roots with 5 cm of length in average, which were then inoculated with M. arenaria. For improved results, plant transformation, and nematode inoculation parameters were adjusted, such as bacterial cell density and growth stage; moist chamber conditions and nematode inoculum concentration. Using this methodology, a candidate gene for nematode resistance, AdEXLB8, was successfully overexpressed in hairy roots of the nematode-susceptible peanut cultivar ‘Runner’, resulting in 98% reduction in the number of galls and egg masses compared to the control, 60 days after M. arenaria infection. Conclusions This methodology proved to be more practical and cost-effective for functional validation of peanut candidate genes than in vitro and composite plant approaches, as it requires less space, reduces analysis costs and displays high transformation efficiency. The reduction in the number of RKN galls and egg masses in peanut hairy roots overexpressing AdEXLB8 corroborated the use of this strategy for functional characterization of root expressing candidate genes. This approach could be applicable not only for peanut–nematode interaction studies but also to other peanut root diseases, such as those caused by fungi and bacteria, being also potentially extended to other crop species displaying similar petiole-rooting competence.

The objective of this work was to evaluate the effect on the physiological and biochemical aspects of garlic submitted to water deficit, knowing that the popular garlic Allium sativum L. is a culture known worldwide for its medicinal, culinary and ecological properties of agricultural pests. were constituted with water condition of 100; 75; 50 and 25% of pot capacity (CP). Three bulbs were planted per pot (5 L) containing Plantmax® substrate, irrigated every two days, according to pot capacity. The chlorophyll index was evaluated during the first and last week of greenhouse treatments. At 45 days after the treatments, the garlic plants were harvested to evaluate the physiological and biochemical parameters. Garlic plants showed to be sensitive to water deficit, resulting in lower leaf length and fresh biomass. Water stress induced the activity of the APX enzyme in the leaves and of the POD in the roots, as well as increase of proteins in the bulbilhos as biochemical mechanisms of adaptation.

Mayaro virus (MAYV) is an arbovirus that circulates in Latin America and is emerging as a potential threat to public health. Infected individuals develop Mayaro fever, a severe inflammatory disease characterized by high fever, rash, arthralgia, myalgia and headache. The disease is often associated with a prolonged arthralgia mediated by a chronic inflammation that can last months. Although the immune response against other arboviruses, such as chikungunya virus (CHIKV), dengue virus (DENV) and Zika virus (ZIKV), has been extensively studied, little is known about the pathogenesis of MAYV infection. In this study, we established models of MAYV infection in macrophages and in mice and found that MAYV can replicate in bone marrow-derived macrophages and robustly induce expression of inflammasome proteins, such as NLRP3, ASC, AIM2, and Caspase-1 (CASP1). Infection performed in macrophages derived from Nlrp3–/–, Aim2–/–, Asc–/–and Casp1/11–/–mice indicate that the NLRP3, but not AIM2 inflammasome is essential for production of inflammatory cytokines, such as IL-1β. We also determined that MAYV triggers NLRP3 inflammasome activation by inducing reactive oxygen species (ROS) and potassium efflux. In vivo infections performed in inflammasome-deficient mice indicate that NLRP3 is involved with footpad swelling, inflammation and pain, establishing a role of the NLRP3 inflammasome in the MAYV pathogenesis. Accordingly, we detected higher levels of caspase1-p20, IL-1β and IL-18 in the serum of MAYV-infected patients as compared to healthy individuals, supporting the participation of the NLRP3-inflammasome during MAYV infection in humans., Author summary Viruses transmitted by mosquitoes have recently received huge attention from the media because the epidemics caused by Zika and chikungunya virus rapidly spread to new areas and infected a large number of individuals. Mayaro is a virus transmitted by mosquitoes that circulates in the Caribbean and tropical regions of Latin America. It causes a highly inflammatory disease, called Mayaro fever, and acute disease is often followed by a prolonged arthralgia mediated by chronic inflammation that can last months or years. The spread of Mayaro to urban areas is a major concern by the authorities, given that the virus has potential to cause an epidemic if spread in high-risk areas. Thus, understanding the mechanisms related to the pathogenesis of this infectious agent would be of great value to treat and prevent the disease. Here, we established an adult mouse model of Mayaro infection and demonstrated that the virus activates the NLRP3 inflammasome, which is important to regulate this viral disease. Our study provides molecular targets for a future treatment of Mayaro fever and provides an experimental model to understand the pathology caused by this emerging viral pathogen.

BRS Esteio is a common bean cultivar with black grains of excellent cooking quality, suited for cultivation in 21 Brazilian States. In 77 trials of Value for Cultivation and Use, the cultivar had a normal cycle (90 days), an average yield of 2,529 kg ha-1, which is 8.1% higher than that of the control cultivars, a high yield potential (4,702 kg ha-1), upright growth, tolerance to lodging, and moderate resistance to anthracnose.

Drought stress, particularly during the flowering and grain-filling stages of growth, contributes to serious yield loss in common bean (Phaseolus vulgaris L.). The aim of this study was to identify genes induced in response to drought stress using transcriptome analysis of contrasting genotypes. Using leaf tissues of tolerant (BAT 477) and susceptible common bean genotypes (Perola), collected at the flowering and grain-filling stages, four complementary deoxyribonucleic acid representational difference analysis subtractive libraries were constructed and then sequenced. A total of 7,203 (77.6 %) sequences with an average sequence size of 570 bp were considered valid, for a combined 4 Mbp sequence. According to a differential display analysis, 802 expressed sequence tags, distributed across 67 contigs, were differentially expressed by the tolerant (37 contigs) and susceptible genotypes (30 contigs) after identification under drought conditions during the two investigated plant developmental stages. Of these differential contigs, the 13 most frequent genes were exclusive to the tolerant genotype. Based on BLAST2GO, 73 % of the gene sequences were annotated and 12 % showed mapping results, with the highest similarity rate corresponding to Glycine max (41 %). According to gene ontology functional analysis, 48 % of the sequences were attributed to cell metabolic processes. Overall, 8.3 % of the transcribed sequences exhibited similarity to transcription factors, predominantly those of the AP2-EREBP family (97.8 %). Of the target sequences validated by quantitative real-time polymerase chain reaction, most genes showed an expression level that agreed with that predicted by in silico analysis. Thus, the drought transcriptome dataset is a valuable resource on the variation in these gene sequences, offering the opportunity to identify robust molecular markers tightly linked to trait-controlling loci for use in marker-assisted breeding.

Cosmopolitan pests such asBrevicoryne brassicae, Lipaphis pseudobrassicae, andMyzus persicae(Aphididae) cause significant damage to Brassicaceae crops. Assessment of the important biotic and abiotic factors that regulate these pests is an essential step in the development of effective Integrated Pest Management programs for these aphids. This study evaluated the influence of leaf position, precipitation, temperature, and parasitism on populations ofL. pseudobrassicae, M. persicae, andB. brassicaein collard greens fields in the Triângulo Mineiro region (Minas Gerais state), Brazil. Similar numbers ofB. brassicaewere found on all parts of the collard green plants, whereasM. persicaeandL. pseudobrassicaewere found in greatest numbers on the middle and lower parts of the plant. While temperature and precipitation were positively related to aphid population size, their effects were not accumulative, as indicated by a negative interaction term. AlthoughDiaeretiella rapaewas the main parasitoid of these aphids, hyperparasitism was dominant; the main hyperparasitoid species recovered from plant samples wasAlloxysta fuscicornis. Parasitoids seem to have similar distributions on plants as their hosts. These results may help predict aphid outbreaks and gives clues for specific intra-plant locations when searching for and monitoring aphid populations.

Breeding efforts to develop peanut (Arachis hypogaea L.) cultivars with multiple resistances, high seed quality and yield have been hindered by the narrow genetic base of the cultivated genepool and the complex nature of its genome. Conversely, peanut wild relatives (Arachis spp.) are considered important sources of resistance alleles, since they have high genetic diversity and have been selected during evolution in a range of adverse environments and conditions. Transcriptome studies on wild species constitute important assets for the identification of genomic segments of interest for transfer into cultivated species. An unprecedented amount of genomic information for wild and cultivated Arachis has been produced in recent years, leading to the discovery of genes and regulatory sequences, and enlarging the collections of molecular markers. The increasing availability of Arachis transcriptomic resources such as ESTs, Unigenes, full-length cDNA clones and derived proteins is enabling a more precise correlation of genotype/phenotype in the genus, with the potential to facilitate accurate intervention in pathways to improve peanut agronomical traits. To maximize these valuable assets, candidate gene validation and peanut genetic transformation methods have been developed to facilitate the deployment of wild alleles into new cultivars.

Cultivated peanut is an allotetraploid (genome type AABB) with a very narrow genetic base, therefore wild species are an attractive source of new variability and traits. Because most wild species are diploid, the first step of introgression usually involves hybridization of wild species and polyploidization to produce a synthetic allotetraploid (AABB) that is sexually compatible with peanut. This study investigates drought-related traits such as leaf morphology, transpiration profile, chlorophyll meter readings (SCMR), specific leaf area (SLA) and transpiration rate per leaf area for two wild diploids ( Arachis duranensis and Arachis ipaensis ) that could be of interest for improvement of the peanut crop. Furthermore, the inheritance of the traits from the diploid to the tetraploid state was investigated. Results showed that whilst some diploid traits such as SCMR, are maintained through hybridization and polyploidization, most characters, such as the leaf area, stomata size, trichome density and transpiration profile, are substantially modified. The study concludes that direct evaluations of drought-related traits in wild diploids may be useful for evaluation of wild species to be used in introgression. However, evaluations on wild-derived synthetic tetraploids are likely to be more informative.

Alzheimer's dementia (AD) is a degenerative brain disorder characterized mainly by cholinergic failure, but other neuro-transmitters are also deficient especially at late stages of the disease. Misfolded β-amyloid peptide has been identified as a causative agent, however inflammatory changes also play a pivotal role. Even though the most prominent pathology is seen in the cognitive functions, specific abnormalities of the central nervous system (CNS) are also reflected in the periphery, particularly in the immune responses of the body. The aim of this study was to characterize the dopaminergic and serotonergic systems in AD, which are also markedly disrupted along with the hallmark acetyl-choline dysfunction. Peripheral blood mono-nuclear cells (PBMCs) from demented patients were judged against comparison groups including individuals with late-onset depression (LOD), as well as non-demented and non-depressed subjects. Cellular sub-populations were evaluated by mono-clonal antibodies against various cell surface receptors: CD4/CD8 (T-lymphocytes), CD19 (B-lymphocytes), CD14 (monocytes), and CD56 (natural-killer (NK)-cells). The expressions of dopamine D(3) and D(4), as well as serotonin 5-HT(1A), 5-HT(2A), 5-HT(2B) and 5-HT(2C) were also assessed. There were no significant differences among the study groups with respect to the frequency of the cellular sub-types, however a unique profound increase in 5-HT(2C) receptor exclusively in NK-cells was observed in AD. The disease-specific expression of 5-HT(2C), as well as the NK-cell cyto-toxicity, has been linked with cognitive derangement in dementia. These changes not only corroborate the existence of bi-directional communication between the immune system and the CNS, but also elucidate the role of inflammatory activity in AD pathology, and may serve as potential biomarkers for less invasive and early diagnostic purposes as well.

Cultivated peanut is an allotetraploid with an AB-genome. In order to learn more of the genomic structure of peanut, we characterized and studied the evolution of a retrotransposon originally isolated from a resistance gene analog (RGA)-containing bacterial artificial chromosome (BAC) clone. It is a moderate copy number Ty1- copia retrotransposon from the Bianca lineage and we named it Matita. Fluorescent in situ hybridization (FISH) experiments showed that Matita is mainly located on the distal regions of chromosome arms and is of approximately equal frequency on both A- and B-chromosomes. Its chromosome-specific hybridization pattern facilitates the identification of individual chromosomes, a useful cytogenetic tool considering that chromosomes in peanut are mostly metacentric and of similar size. Phylogenetic analysis of Matita elements, molecular dating of transposition events, and an estimation of the evolutionary divergence of the most probable A- and B-donor species suggest that Matita underwent its last major burst of transposition activity at around the same time of the A- and B-genome divergence about 3.5 million years ago. By probing BAC libraries with overgos probes for Matita, resistance gene analogues, and single- or low-copy genes, it was demonstrated that Matita is not randomly distributed in the genome but exhibits a significant tendency of being more abundant near resistance gene homologues than near single- copy genes. The described work is a further step towards broadening the knowledge on genomic and chromosomal structure of peanut and on its evolution.

Peanut (Arachis hypogaea) is amongst the most important legume crops in the world. One of its main yield constraints is the root-knot nematode Meloidogyne arenaria. A number of wild Arachis species, including A. stenosperma, are resistant to nematodes, and are a potential source of new resistance alleles for cultivated peanut. Using in silico subtraction of ESTs and macroarray analysis, we identified genes differentially expressed in A. stenosperma roots during its resistance response to M. arenaria. The three most differentially expressed genes [Auxin Repressed Protein (AsARP), Cytokinin Oxidase (AsCKX) and Metallothionein Type 2 (AsMET2)] were further analyzed using northern-blot and showed distinct expression profiles in the resistant A. stenosperma and susceptible A. hypogaea, both after, and sometimes even before, challenge with nematodes. Of the three most differentially expressed genes, AsARP and AsCKX are potentially involved in plant hormonal balance, and AsMET2 may be related to the reactive oxygen reaction triggered by the hypersensitive response (HR).

It is well known that dopamine imbalances are associated with many psychiatric disorders and that the dopaminergic receptor D₂ is the main target of antipsychotics. Recently it was shown that levels of two proteins implicated in dopaminergic signaling, Neuronal calcium sensor-1 (NCS-1) and DARPP-32, are altered in the prefrontal cortex (PFC) of both schizophrenic and bipolar disorder patients. NCS-1, which inhibits D₂ internalization, is upregulated in the PFC of both patients. DARPP-32, which is a downstream effector of dopamine signaling, integrates the pathways of several neurotransmitters and is downregulated in the PFC of both patients. Here, we used PC12 cells stably overexpressing NCS-1 (PC12-NCS-1 cells) to address the function of this protein in DARPP-32 signaling pathway in vitro. PC12-NCS-1 cells displayed downregulation of the cAMP/PKA pathway, with decreased levels of cAMP and phosphorylation of CREB at Ser133. We also observed decreased levels of total and phosphorylated DARPP-32 at Thr34. However, these cells did not show alterations in the levels of D₂ and phosphorylation of DARPP-32 at Thr75. These results indicate that NCS-1 modulates PKA/cAMP signaling pathway. Identification of the cellular mechanisms linking NCS-1 and DARPP-32 may help in the understanding the signaling machinery with potential to be turned into targets for the treatment of schizophrenia and other debilitating psychiatric disorders.

Cultivated peanut, Arachis hypogaea L., is an economically important species. It is very susceptible to different stresses to which wild species are mostly resistant. Foliar diseases, such as late leaf spot (LLS) caused by the fungus Cercosporidium personatum, and rust caused by the fungus Puccinia arachidis, are responsible for decrease in plant growth and productivity. The peanut wild relative Arachis stenosperma accession V 10309 was identified as resistant to a number of pests and diseases, including LLS and rust. Aiming to better understand the mechanisms of resistance of A. stenosperma to C. personatum and P. arachidis, determine initial key steps of the plant-pathogen interaction and to contribute for studies on genes involved in this interaction, ultrastructural analysis was performed on leaves of A. stenosperma V10309 (wild, resistant) and A. hypogaea cv. IAC-Tatu (cultivated, susceptible) inoculated with C. personatum or P. arachidis. For both fungal species, adhesion, germination of spores and hyphal proliferation occurred in both species but was more limited and later in A. stenosperma than in A. hypogaea, and no successful penetration was observed in the former. These data suggest that in A. stenosperma, infection is hampered at the stage of penetration. This is the first morphological description of the first hours of the interaction of plant pathogenic fungi and the resistant wild species A. stenosperma.

Essential plant nutrients are mainly applied to soil and plant foliage for achieving maximum economic yields. Soil application method is more common and most effective for nutrients, which required in higher amounts. However, under certain circumstances, foliar fertilization is more economic and effective. Foliar symptoms, soil and plant tissue tests, and crop growth responses are principal nutrient disorder diagnostic techniques. Soil applications of fertilizers are mainly done on the basis of soil tests, whereas foliar nutrient applications are mainly done on the basis of visual foliar symptoms or plant tissue tests. Hence, correct diagnosis of nutrient deficiency is fundamental for successful foliar fertilization. In addition, there are some more requirements for successful foliar fertilization. Foliar fertilization requires higher leaf area index for absorbing applied nutrient solution in sufficient amount, it may be necessary to have more than one application depending on severity of nutrient deficiency. Nutrient concentration and day temperature should be optimal to avoid leaf burning and fertilizer source should be soluble in water to be more effective. Foliar fertilization of crops can complement soil fertilization. If foliar fertilization is mixed with postemergence herbicides, insecticides, or fungicides, the probability of yield response could be increased and cost of application can be reduced.

The reproductive behaviour of the root-knot nematode Meloidogyne arenaria race 1 was compared on two wild species of Arachis (A. duranensis and A. stenosperma) and cultivated peanut (A. hypogaea cv. IAC-Tatu-ST). The three species were considered moderately susceptible, resistant, and susceptible, respectively. Penetration and development of the root-knot nematode in the resistant species was reduced in comparison with that occurring in susceptible plants. Several cell features, including dark blue cytoplasm and altered organelle structure were observed in the central cylinder of A. stenosperma, indicating a hypersensitive-like response (HR) of infested host cells. Neither giant cells, nor nematodes developed beyond the second stage, were found on A. stenosperma. Arachis duranensis showed a delay in the development of nematodes in the roots compared to A. hypogaea. The two wild peanut species were chosen to be the contrasting parents of a segregating population for mapping and further investigation of resistance genes.

Peanut is an important legume crop. Although it has high levels of morphological diversity, it lacks genetic variability and sources of disease resistance. The transference of resistance genes from wild species is difficult due to the different ploidy level of the wild and cultivated species. Recently, amphidiploids have been produced that can be used as 'bridges' to introgress wild genes. Molecular markers are useful to pyramidize desirable genes and track them through generations of backcrossings. Molecular markers based on Resistance Gene Analogs have improved chances to be present in or linked to resistance gene loci. This study describes the development and genotyping of molecular markers based on resistance gene motifs. Specific primers were designed based on unique sequences of an Arachis RGA dataset. The identity of the amplified polymorphic bands was confirmed by sequencing. These markers were genotyped on a F2 population that segregates for resistance to biotic stress types.

Root-knot nematodes (RKN; Meloidogyne sp.) are a major threat to crops in tropical and subtropical regions worldwide. The use of resistant crop varieties is the preferred method of control because nematicides are expensive, and hazardous to humans and the environment. Peanut (Arachis hypogaea) is infected by four species of RKN, the most damaging being M. arenaria, and commercial cultivars rely on a single source of resistance. In this study, we genetically characterize RKN resistance of the wild Arachis species A. stenosperma using a population of 93 recombinant inbred lines developed from a cross between A. duranensis and A. stenosperma. Four quantitative trait loci (QTL) located on linkage groups 02, 04, and 09 strongly influenced nematode root galling and egg production. Drought-related, domestication and agronomically relevant traits were also evaluated, revealing several QTL. Using the newly available Arachis genome sequence, easy-to-use KASP (kompetitive allele specific PCR) markers linked to the newly identified RKN resistance loci were developed and validated in a tetraploid context. Therefore, we consider that A. stenosperma has high potential as a new source of RKN resistance in peanut breeding programs.

Cultivated peanut (Arachis hypogaea) is an allotetraploid with closely related subgenomes of a total size of ∼2.7 Gb. This makes the assembly of chromosomal pseudomolecules very challenging. As a foundation to understanding the genome of cultivated peanut, we report the genome sequences of its diploid ancestors (Arachis duranensis and Arachis ipaensis). We show that these genomes are similar to cultivated peanut's A and B subgenomes and use them to identify candidate disease resistance genes, to guide tetraploid transcript assemblies and to detect genetic exchange between cultivated peanut's subgenomes. On the basis of remarkably high DNA identity of the A. ipaensis genome and the B subgenome of cultivated peanut and biogeographic evidence, we conclude that A. ipaensis may be a direct descendant of the same population that contributed the B subgenome to cultivated peanut.

A higher prevalence of Strongyloides stercoralis infections has been reported in alcoholic patients compared to nonalcoholic patients living in the same area. Excessive alcohol consumption increases the levels of endogenous corticosteroids that subsequently enhance the fecundity of S. stercoralis parthenogenetic females. These corticosteroids also enhance the transformation of rhabditiform larvae into infective filariform larvae by mimicking the effect of the ecdysteroid hormones produced by the parasite, thus leading to autoinfection. In addition, alterations in the intestinal barrier and host immune response contribute to the development of hyperinfection and severe strongyloidiasis in alcoholic patients. The aim of this study was to evaluate the frequency of S. stercoralis infections in alcoholic patients and to determine the association between S. stercoralis infection and endogenous cortisol levels. The frequency of infection was evaluated in 332 alcoholic and 92 nonalcoholic patients. The parasitological diagnosis was carried out by agar plate culture, the modified Baermann-Moraes method and spontaneous sedimentation. The immunological diagnosis was performed using an ELISA with anti-S. stercoralis IgG. The cortisol levels were measured in serum samples by ELISA. The frequency of S. stercoralis infection in alcoholic patients was 23.5% (78/332), while in nonalcoholic patients, it was 5.4% (5/92) (p0.05). The cortisol levels were higher in alcoholic than in nonalcoholic patients (p0.05). However, among the alcoholic patients, the cortisol levels did not differ between S. stercoralis-infected and uninfected patients (p0.05). As demonstrated in this work, 81.3% (26/32) of patients with a high parasite load, considered as more than 11 larvae per gram of feces, presented serum cortisol levels above the normal reference value (24 mg/dL). High endogenous cortisol levels in alcoholic patients were not associated to susceptibility to S. stercoralis infection, however once infected, this may lead to a high parasite load.

Changes in dopaminergic function can be regulated by receptor-receptor interaction, or interaction with other proteins with dopamine receptors, and/or elements of the downstream signaling cascades. The complexity of dopaminergic signaling is far from being completely elucidated. It could, however, hold the key to the comprehension of the pathophysiology of neurological and psychiatric disorders, as well as to the identification of putative new targets for, and development of, more efficacious and selective drugs. Here, we review some of the current evidence and new ideas that are being proposed as a result, as well as future perspectives that are now being recognized.

Despite the availability of several Saccharomyces cerevisiae commercial strains intended for wine production, strains isolated from winery regions are usually more adapted to their own climatic conditions, grapes and also partially responsible for particular characteristics that frequently identify specific wines and regions. Thus the microbiota of an important winery region (Colombo) was studied in order to isolate and characterize S. cerevisiae strains that could be used on wine production. From 61 yeasts isolated, 14 were identified as S. cerevisiae. Some of them showed fermentative characteristics even better than commercial strains indicating that they could be applied on wine production in order to increase the quality and assure the particular wine characteristics of that region.

O estabelecimento de bancos de sementes para conservação ex situ de germoplasma vegetal é largamente utilizado, mas a contaminação das mesmas por fitopatógenos pode comprometer sua integridade. Este trabalho teve por objetivo monitorar a sobrevivência de Xanthomonas axonopodis pv. phaseoli, em um lote de sementes de feijão (Phaseolus vulgaris) cv. Roxão)submetido a três condições de temperatura, durante 60 meses de armazenamento: -18 e 5 ºC, condições preconizadas para a conservação de material genético vegetal a longo e médio prazo, respectivamente, e à temperatura ambiente (20-30 ºC). A sobrevivência da bactéria foi avaliada pela percentagem de sementes contaminadas e a viabilidade pela manutenção da patogenicidade dos isolados. A população de X. axonopodis pv. phaseoli nas sementes foi quantificada ao final de cinco anos de armazenamento. Os resultados mostraram que a percentagem de sementes contaminadas decresceu de 64 para 36-37% nos primeiros seis meses, para os três tratamentos. A partir de 30 meses observou-se que as sementes conservadas a -18 e 5 ºC apresentaram níveis de contaminação (58 e 72%) que diferiram significativamente das conservadas à temperatura ambiente (20%), e aos 60 meses taxas de 46%, 46% e 8%, respectivamente. A temperatura mais propícia à sobrevivência da bactéria foi 5 ºC em que se encontrou uma população máxima de 1,2 x 10(8) ufc/semente. Constatou-se, igualmente, a manutenção do poder infetivo dos isolados durante todo o armazenamento. Pode-se concluir que as condições ótimas para a conservação de sementes são as mesmas para a manutenção da longevidade de X. axonopodis pv. phaseoli.

Summary The protein Tarin 1, from Colocasia esculenta, was expressed in Nicotiana tabacum. Bioassays were done on plants expressing Tarin 1 at different levels using Spodoptera frugiperda larvae, various bacteria and fungi and the root-knot nematode Meloidogyne javanica. It was found that S. frugiperda larvae fed on transformed plants had retarded and lower pupation, lower accumulated biomass and higher mortality rate than larvae fed on control plants. Also, Tarin 1 was found to inhibit the growth in vitro of Pseudomonas syringae pv. tomato. For Meloidogyne javanica, both relative replication and root damage were greater in control plants than in transformed plants, but the results were not statistically significant. This work illustrates the effects of plants expressing Tarin 1, on the growth and development of insects and bacteria, and shows its potential for pest management.

We conducted a study of the basal levels of gonadotropins in 38 patients with Turner's syndrome ,14 of whom were using hormone replacement therapy (HRT). The gonadotropin levels were compared with pubertal development and HRT. Seven patients had presented spontaneous menarche; five patients maintained their periods and normal gonadotropin levels ,and two developed secondary amenorrhea and high gonadotropin levels. The majority of patients on HRT had high gonadotropin levels. Follicle stimulating hormone (FSH) levels were significantly lower (p = 0.001) in patients with breast development at stage 5, regardless of whether the patient had undergone a spontaneous or a hormonally induced puberty. We concluded that gonadotropin levels are normal in those patients with spontaneous periods ,and are high in most patients on HRT; and that FSH levels are significantly lower in those patients with breasts at stage 5.

Avaliamos em 26 meninas com pubarca precoce (PP) e 14 controles (7,3±1,0 x 7,0±2,0 anos) os efeitos dos androgênios adrenais e periféricos na modulação dos níveis da globulina ligadora dos hormônios sexuais (SHBG) na infância. Foram avaliados: a idade óssea (IO) e o índice de massa corporal (IMC) e os níveis de androstenediona (A), testosterona (T), dehidro-epiandrostenediona e seu sulfato (DHEA e SDHEA), dihidrotestosterona (DHT), 3a-androstanediol glucuronídeo (3a-diolG) e SHBG. A IO (8,0±1,4 x 8,3±1,4 anos) foi semelhante, mas o IMC (18,7±3,1 x 16,7±2,0, p=0,03) foi maior na PP. O SDHEA (0,7±0,4 x 0,3±0,2µg/mL, p