Enraizamento de microestacas de mirtileiro provenientes de microjardim clonal semi-hidropônico

O uso de microjardins clonais hidropônicos tem sido relatado com sucesso para espécies florestais e pode vir a se tornar uma excelente alternativa para espécies frutíferas de difícil propagação, como é o caso do mirtilo. O objetivo deste estudo foi avaliar o enraizamento de microestacas de mirtileiro provenientes de dois sistemas de cultivo (convencional e semi-hidropônico), submetidas a diferentes concentrações de AIB (ácido indolbutírico). As microestacas de mirtileiro das cultivares Bluebelle e Woodard foram submetidas a diferentes concentrações de AIB (0; 500; 1.000; 1.500 e 2.000 mg.L-1), acondicionadas em caixas plásticas contendo vermiculita e, aos 90 dias de cultivo, avaliou-se o seu rendimento. O delineamento experimental foi o inteiramente casualizado, com vinte tratamentos, contendo quatro repetições, compostas por dez microestacas cada. Foram avaliados as porcentagem de sobrevivência e de enraizamento, o comprimento da maior raiz, o número de brotações, o comprimento médio das brotações, o número de folhas e as massas fresca e seca radiculares. O sistema semi-hidropônico proporcionou um rendimento de microestacas significativamente superior ao convencional; entretanto, este material apresentou menores porcentagens de sobrevivência e enraizamento. Todos os tratamentos, inclusive aquele sem a presença de AIB, apresentaram porcentagens de enraizamento superiores a 50%.

Influência genotípica e clonal de genótipos de bananeiras micropropagados a partir de gemas florais masculinas

Inflorescências masculinas de bananeiras apresentam potencial para serem usadas como explantes para a micropropagação de bananeiras. Este trabalho teve por objetivo avaliar a influência genotípica e clonal de bananeiras micropropagadas a partir de gemas florais masculinas em sucessivos subcultivos. Foram utilizados explantes florais de 15 clones plantados em campo, da variedade Preciosa e do híbrido FHIA 02, em meio de cultura de MS contendo 4,0 mg.L-1 de BAP. Por sete subcultivos sucessivos de 30 dias, os materiais foram avaliados quanto à taxa de multiplicação e contaminação microbiana. Ao final do período de multiplicação, o acúmulo de mudas produzidas, em razão dos sucessivos subcultivos utilizando esse tipo de explante, foi também avaliado. Verificou-se que os maiores índices de contaminação foram observados no primeiro subcultivo, com média superior a 50% de contaminação dos explantes. As melhores taxas de multiplicação foram alcançadas no terceiro subcultivo, com média de até 6,1 brotos por explante. Os clones que apresentaram maior acúmulo de mudas produziram 105 e 163 mudas por explante, para a variedade Preciosa e o híbrido FHIA 2, respectivamente, após sete subcultivos.

Eosinophilic fasciitis: Typical abnormalities, variants and differential diagnosis of fasciae abnormalities using MR imaging.

Eosinophilic fasciitis is a rare condition. It is generally limited to the distal parts of the arms and legs. MRI is the ideal imaging modality for diagnosing and monitoring this condition. MRI findings typically evidence only fascial involvement but on a less regular basis signal abnormalities may be observed in neighboring muscle tissue and hypodermic fat. Differential diagnosis of eosinophilic fasciitis by MRI requires the exclusion of several other superficial and deep soft tissue disorders.

Limpeza clonal de mudas de videira infectadas por Xanthomonas campestris pv. viticola

O cancro bacteriano da videira é causado por Xanthomonas campestris pv. viticola (Xcv). Visando à limpeza clonal de mudas de 'Red Globe', foram estudados: tamanho ideal de ápices e gemas axilares para cultivo em meio de Galzy modificado (MGM); efeito da termoterapia (38ºC/30 dias); e ação de antibióticos na eliminação de Xcv em videiras infectadas. Os percentuais de contaminação por Xcv e de regeneração foram analisados, e as plantas obtidas foram indexadas em meio ágar nutritivo-dextrose-extrato de levedura-ampicilina (NYDAM), seguindo-se teste de patogenicidade. O cultivo de explantes com 3 mm possibilitou a obtenção de plantas livres da bactéria, com regeneração 14,3 vezes maior que explantes com 1 mm. A termoterapia de mudas infectadas, associada ao cultivo in vitro, não eliminou o patógeno. O cultivo de explantes com 10 mm, durante 40 dias em MGM + cefotaxima (300 mg L-1), proporcionou limpeza clonal das mudas. A indexação de plantas de videira regeneradas in vitro, quanto à infecção por Xcv utilizando NYDAM, seguida de teste de patogenicidade, é uma alternativa econômica e eficiente para produção de mudas de alta qualidade fitossanitária.

MICROJARDIM CLONAL DE MIRTILEIRO EM SISTEMAS DE CULTIVO SEM SOLO

RESUMO O uso de microjardins clonais em sistemas de cultivo sem solo para fornecimento de material propagativo na cultura do mirtileiro (Vaccinium spp.) pode trazer grandes avanços na produção de mudas dessa cultura. O objetivo desta pesquisa foi avaliar a produção de microestacas e a sobrevivência de plantas matrizes de mirtileiro micropropagadas das cultivares Woodard e Aliceblue, em dois sistemas de cultivo. Os sistemas de cultivo utilizados foram o semi-hidropônico (floreiras com substrato de areia e fornecimento diário de solução nutritiva) e o com substrato organomineral (sacos plásticos com substrato comercial e fornecimento de solução nutritiva a cada 15 dias). Após o período de 90 dias do plantio das plantas matrizes, foram iniciadas as coletas de microestacas, as quais foram realizadas a cada 60 dias, com exceção do período de inverno, em que as coletas foram realizadas a cada 90 dias, totalizando ao final do experimento onze coletas. O experimento foi constituído como um fatorial 2 x 2 x 11 (sistemas x cultivares x coletas), em delineamento inteiramente casualizado, com três repetições de 12 plantas cada. Foram avaliadas a produção total de microestacas ao final das onze coletas, o número de microestacas produzidas por planta matriz a cada coleta, a sobrevivência das plantas matrizes ao final das onze coletas e a sobrevivência das plantas matrizes a cada coleta. Os resultados indicaram que o sistema semi-hidropônico foi superior ao substrato organomineral para a produção de microestacas de ambas as cultivares. A maior produtividade total de microestacas ocorreu no sistema semi-hidropônico combinado com a cultivar Aliceblue, com produção total média de 237,67 microestacas. Porém, nesta condição, houve menor sobrevivência das plantas matrizes. A produção de microestacas apresentou alternância ao longo das coletas. A sobrevivência das plantas matrizes diminuiu após sucessivas coletas. Após as coletas de verão, ocorreu maior mortalidade de plantas matrizes.

A systems biology approach identifies molecular networks defining skeletal muscle abnormalities in chronic obstructive pulmonary disease.

Chronic Obstructive Pulmonary Disease (COPD) is an inflammatory process of the lung inducing persistent airflow limitation. Extensive systemic effects, such as skeletal muscle dysfunction, often characterize these patients and severely limit life expectancy. Despite considerable research efforts, the molecular basis of muscle degeneration in COPD is still a matter of intense debate. In this study, we have applied a network biology approach to model the relationship between muscle molecular and physiological response to training and systemic inflammatory mediators. Our model shows that failure to co- ordinately activate expression of several tissue remodelling and bioenergetics pathways is a specific landmark of COPD diseased muscles. Our findings also suggest that this phenomenon may be linked to an abnormal expression of a number of histone modifiers, which we discovered correlate with oxygen utilization. These observations raised the interesting possibility that cell hypoxia may be a key factor driving skeletal muscle degeneration in COPD patients.

Short stature and primary ovarian insufficiency possibly due to chromosomal position effect in a balanced X;1 translocation.

BACKGROUND: Primary ovarian insufficiency (POI) is defined as a primary ovarian defect characterized by absent menarche (primary amenorrhea), a decrease in the initial primordial follicle number, high follicle-stimulating hormone (FSH) levels and hypoestrogenism. Although the etiology of a majority of POI cases is not yet identified, several data suggest that POI has a strong genetic component. Conventional cytogenetic and molecular analyses have identified regions of the X chromosome that are associated with ovarian function, as well as POI candidate genes, such as FMR1 and DIAPH2. Here we describe a 10.5-year-old girl presenting with high FSH and luteinizing hormone (LH) levels, pathologic GH stimulation arginine and clonidine tests, short stature, pterygium, ovarian dysgenesis, hirsutism and POI. RESULTS: Cytogenetic analysis demonstrated a balanced reciprocal translocation between the q arms of chromosomes X and 1, with breakpoints falling in Xq21 and 1q41 bands. Molecular studies did not unravel any chromosome microdeletion/microduplication, and no XIST-mediated inactivation was found on the derivative chromosome 1. Interestingly, through immunofluorescence assays, we found that part of the Xq21q22 trait, translocated to chromosome 1q41, was late replicating and therefore possibly inactivated in 30 % metaphases both in lymphocytes and skin fibroblasts, in addition to a skewed 100 % inactivation of the normal X chromosome. These findings suggest that a dysregulation of gene expression might occur in this region. Two genes mapping to the Xq translocated region, namely DIAPH2 and FMR1, were found overexpressed if compared with controls. CONCLUSIONS: We report a case in which gonadal dysgenesis and POI are associated with over-expression of DIAPH2 gene and of FMR1 gene in wild type form. We hypothesize that this over-expression is possibly due to a phenomenon known as "chromosomal position effect", which accounts for gene expression variations depending on their localization within the nucleus. For the same effect a double mosaic inactivation of genes mapping to the Xq21-q22 region, demonstrated by immunofluorescence assays, may be the cause of a functional Xq partial monosomy leading to most Turner traits of the proband's phenotype.

Dendritic Spine Abnormalities in Hippocampal CA1 Pyramidal Neurons Underlying Memory Deficits in the SAMP8 Mouse Model of Alzheimer's Disease

SAMP8 is a strain of mice with accelerated senescence. These mice have recently been the focus of attention as they show several alterations that have also been described in Alzheimer"s disease (AD) patients. The number of dendritic spines, spine plasticity, and morphology are basic to memory formation. In AD, the density of dendritic spines is severely decreased. We studied memory alterations using the object recognition test. We measured levels of synaptophysin as a marker of neurotransmission and used Golgi staining to quantify and characterize the number and morphology of dendritic spines in SAMP8 mice and in SAMR1 as control animals. While there were no memory differences at 3 months of age, the memory of both 6- and 9-month-old SAMP8 mice was impaired in comparison with age-matched SAMR1 mice or young SAMP8 mice. In addition, synaptophysin levels were not altered in young SAMP8 animals, but SAMP8 aged 6 and 9 months had less synaptophysin than SAMR1 controls and also less than 3-month-old SAMP8 mice. Moreover, while spine density remained stable with age in SAMR1 mice, the number of spines started to decrease in SAMP8 animals at 6 months, only to get worse at 9 months. Our results show that from 6 months onwards SAMP8 mice show impaired memory. This age coincides with that at which the levels of synaptophysin and spine density decrease. Thus, we conclude that together with other studies that describe several alterations at similar ages, SAMP8 mice are a very suitable model for studying AD.

Imaging findings in fetal diaphragmatic abnormalities.

Imaging plays a key role in the detection of a diaphragmatic pathology in utero. US is the screening method, but MRI is increasingly performed. Congenital diaphragmatic hernia is by far the most often diagnosed diaphragmatic pathology, but unilateral or bilateral eventration or paralysis can also be identified. Extralobar pulmonary sequestration can be located in the diaphragm and, exceptionally, diaphragmatic tumors or secondary infiltration of the diaphragm from tumors originating from an adjacent organ have been observed in utero. Congenital abnormalities of the diaphragm impair normal lung development. Prenatal imaging provides a detailed anatomical evaluation of the fetus and allows volumetric lung measurements. The comparison of these data with those from normal fetuses at the same gestational age provides information about the severity of pulmonary hypoplasia and improves predictions about the fetus's outcome. This information can help doctors and families to make decisions about management during pregnancy and after birth. We describe a wide spectrum of congenital pathologies of the diaphragm and analyze their embryological basis. Moreover, we describe their prenatal imaging findings with emphasis on MR studies, discuss their differential diagnosis and evaluate the limits of imaging methods in predicting postnatal outcome.

A chromosomal insertion toolbox for promoter probing, mutant complementation and pathogenicity studies in Ralstonia solanacearum

We describe here the construction of a delivery system for stable and directed insertion of gene constructs in a permissive chromosomal site of the bacterial wilt pathogen Ralstonia solanacearum. The system consists of a collection of suicide vectors the Ralstonia chromosome (pRC) series that carry an integration element flanked by transcription terminators and two sequences of homology to the chromosome of strain GMI1000, where the integration element is inserted through a double recombination event. Unique restriction enzyme sites and a GATEWAY cassette enable cloning of any promoter::gene combination in the integration element. Variants endowed with different selectable antibiotic resistance genes and promoter::gene combinations are described. We show that the system can be readily used in GMI1000 and adapted to other R. solanacearum strains using an accessory plasmid. We prove that the pRC system can be employed to complement a deletion mutation with a single copy of the native gene, and to measure transcription of selected promoters in monocopy both in vitro and in planta. Finally, the system has been used to purify and study secretion type III effectors. These novel genetic tools will be particularly useful for the construction of recombinant bacteria that maintain inserted genes or reporter fusions in competitive situations (i.e., during plant infection).

The genetic content of chromosomal inversions across a wide latitudinal gradient

There is increasing evidence regarding the role of chromosomal inversions in relevant biological processes such as local adaptation and speciation. A classic example of the adaptive role of chromosomal polymorphisms is given by the clines of inversion frequencies in Drosophila subobscura, repeatable across continents. Nevertheless, not much is known about the molecular variation associated with these polymorphisms. We characterized the genetic content of ca. 600 individuals from nine European populations following a latitudinal gradient by analysing 19 microsatellite loci from two autosomes (J and U) and the sex chromosome (A), taking into account their chromosomal inversions. Our results clearly demonstrate the molecular genetic uniformity within a given chromosomal inversion across a large latitudinal gradient, particularly from Groningen (Netherlands) in the north to Málaga (Spain) in the south, experiencing highly diverse environmental conditions. This low genetic differentiation within the same gene arrangement across the nine European populations is consistent with the local adaptation hypothesis for th evolutionof chromosomal polymorphisms. We also show the effective role of chromosomal inversions in maintaining different genetic pools within these inverted genomic regions even in the presence of high gene flow. Inversions represent thus an important barrier to gene flux and can help maintain specific allelic combinations with positive effects on fitness. Consistent patterns of microsatellite allele-inversion linkage disequilibrium particularly in loci within inversions were also observed. Finally, we identified areas within inversions presenting clinal variation that might be under selection.

Robust DNA Methylation in the Clonal Raider Ant Brain.

Social insects are promising model systems for epigenetics due to their immense morphological and behavioral plasticity. Reports that DNA methylation differs between the queen and worker castes in social insects [1-4] have implied a role for DNA methylation in regulating division of labor. To better understand the function of DNA methylation in social insects, we performed whole-genome bisulfite sequencing on brains of the clonal raider ant Cerapachys biroi, whose colonies alternate between reproductive (queen-like) and brood care (worker-like) phases [5]. Many cytosines were methylated in all replicates (on average 29.5% of the methylated cytosines in a given replicate), indicating that a large proportion of the C. biroi brain methylome is robust. Robust DNA methylation occurred preferentially in exonic CpGs of highly and stably expressed genes involved in core functions. Our analyses did not detect any differences in DNA methylation between the queen-like and worker-like phases, suggesting that DNA methylation is not associated with changes in reproduction and behavior in C. biroi. Finally, many cytosines were methylated in one sample only, due to either biological or experimental variation. By applying the statistical methods used in previous studies [1-4, 6] to our data, we show that such sample-specific DNA methylation may underlie the previous findings of queen- and worker-specific methylation. We argue that there is currently no evidence that genome-wide variation in DNA methylation is associated with the queen and worker castes in social insects, and we call for a more careful interpretation of the available data.

Different rates of defense evolution and niche preferences in clonal and nonclonal milkweeds (Asclepias spp.).

Given the dual role of many plant traits to tolerate both herbivore attack and abiotic stress, the climatic niche of a species should be integrated into the study of plant defense strategies. Here we investigate the impact of plant reproductive strategy and components of species' climatic niche on the rate of chemical defense evolution in the milkweeds using a common garden experiment of 49 species. We found that across Asclepias species, clonal reproduction repeatedly evolved in lower temperature conditions, in species generally producing low concentrations of a toxic defense (cardenolides). Additionally, we found that rates of cardenolide evolution were lower for clonal than for nonclonal species. We thus conclude that because the clonal strategy is based on survival, long generation times, and is associated with tolerance of herbivory, it may be an alternative to toxicity in colder ecosystems. Taken together, these results indicate that the rate of chemical defense evolution is influenced by the intersection of life-history strategy and climatic niches into which plants radiate.

Chromosomal polymorphism of D. subobscura: no differences between wild males and sons of wild females

When analyzing the chromosomal polymorphism of D. subobscura natural populations it is assumed that the information provided by wild males and sons of wild females is equivalent. Thus, using both in the analysis it is possible to increase the sample size. However, it is important to verify whether there are significant differences between both groups or not. The aim of this research has been to statistically compare the results of chromosomal polymorphism of both groups. We have used data from Avala Mountain (Serbia) where D. subobscura flies were collected from the 30th May to the 5th June 2011. Avala is located 18 km south of Belgrade and the trapping place is a forest with polydominant communities of Fagetum submontanum Table 1. Number and percentage of adult flies collected in Font Groga (Barcelona, Spain) on 9th October 2013. Males and sons of wild females were crossed with virgin females of the Küsnacht strain. Third instar larvae from F1 were dissected to obtain the salivary glands and the polytene chromosomes were stained and squashed in aceto-orcein solution. No significant differences were observed for any chromosome of the karyotype: A (p-value = 0.485), J (p-value = 0.230), U (p-value =0.572), E (p-value = 0.536), and O (p-value = 0.338). Thus, it seems that the two groups can be grouped together to obtain the chromosomal polymorphism of the population.