Detection and partial molecular characterization of Grapevine fleck virus, Grapevine virus D, Grapevine leafroll-associated virus -5 and -6 infecting grapevines in Brazil.

Nota científica

Produção de antissoro policlonal utilizando a proteína capsidial recombinante do Rupestris stem pitting-associated virus.

O Rupestris stem pitting-associated virus (RSPaV) é o agente causal das caneluras do lenho da videira. Este trabalho teve como objetivo produzir antissoro policlonal a partir da proteína capsidial (CP) recombinante do RSPaV e avaliar a sua especificidade e sensibilidade. O gene da CP do RSPaV, com 780pb, foi previamente caracterizado. Esse gene foi subclonado no sítio de restrição EcoRI, no vetor de expressão pRSET-B e o plasmídeo recombinante foi utilizado para induzir a expressão da CP em Escherichia coli. A CP, ligada a uma cauda de seis histidinas, foi purificada por meio de cromatografia de afinidade em coluna de Ni-NTA a partir do extrato de proteínas totais extraídas de E. coli. A identidade da proteína purificada foi confirmada em SDS-PAGE e Western blot, utilizando-se anticorpos comerciais contra a cauda de seis histidinas. A CP recombinante expressada in vitro apresentou massa molecular de cerca de 31kDa. A proteína purificada foi quantificada e 2,55mg foram utilizados para a imunização de um coelho. O antissoro policlonal obtido reagiu com diferentes isolados deste vírus, extraídos de videiras em ELISA indireto.Nota técnica

Simultaneous detection of Brazilian isolates of grapevine viruses by TaqMan real-time RT-PCR.

The aim of this work was to evaluate the efficiency of real-time RT-PCR for detection of different isolates of ten important virus species that infect grapevines in Brazil: Grapevine leafroll-associated virus (GLRaV-1, -2, -3 and -5), Grapevine virus A (GVA), Grapevine virus B (GVB), Grapevine virus D (GVD), Grapevine rupestris stem pitting-associated virus (GRSPaV), Grapevine fleck virus (GFkV) and Grapevine fanleaf virus (GFLV). The reactions consisted of individual (simplex) and simultaneous (duplex) virus detections. Thirty six grapevine accessions, regenerated after thermotherapy and tissue culture treatments, have been analysed. All the above-mentioned viruses were sensitively detected in simplex reactions in samples infected with different virus isolates. Specifically to GLRaV-1 it was necessary to mix reagents refered by different sources to achieve the amplification. GVA, GRSPaV, GLRaV-2 and GLRaV-3 combined with GVB, GFLV, GFkV, GVD and GLRaV-5 were accurately detected in duplex trials. It was shown, that real-time RT-PCR (TaqMan) is able to efficiently detect different local virus species and isolates.Comunicação científica

Detecção por RT-PCR em Tempo Real e caracterização molecular do Grapevine virus D e Grapevine leafroll-associated virus 5 em videira.

O objetivo deste trabalho foi verificar a ocorrência destes dois vírus em videiras por meio de RT-PCR em Tempo Real, além de caracterizar molecularmente três dos isolados obtidos.Resumo

High-sensitivity monitoring of micromechanical vibration using optical whispering gallery mode resonators

The inherent coupling of optical and mechanical modes in high finesse optical microresonators provide a natural, highly sensitive transduction mechanism for micromechanical vibrations. Using homodyne and polarization spectroscopy techniques, we achieve shot-noise limited displacement sensitivities of 10^(-19) m Hz^(-1/2). In an unprecedented manner, this enables the detection and study of a variety of mechanical modes, which are identified as radial breathing, flexural and torsional modes using 3-dimensional finite element modelling. Furthermore, a broadband equivalent displacement noise is measured and found to agree well with models for thermorefractive noise in silica dielectric cavities. Implications for ground-state cooling, displacement sensing and Kerr squeezing are discussed.Comment: 25 pages, 8 figure

Biological and molecular characterization of two Brazilian isolates of Apple stem grooving virus.

Apple stem grooving virus (ASGV, genus Capillovirus) is disseminated worldwide, usually causing a latent infection in most commercial apple cultivars. However, infected scions grafted onto sensitive material display reduction of yield, loss of fruit quality and tree decline. In Brazil ASGV is associated with severe phloem necrosis, xylem pitting and decline of apple trees on Maruba-kaido (Malus prunifolia cv. Ringo) rootstocks usually in a complex with other latent viruses. Two Brazilian ASGV isolates from a mixed infection causing differing reactions on apple cv. Fuji on Maruba-kaido rootstocks, i.e., normal growth vs. tree decline, have been completely sequenced. The differing intensity of symptoms they co-induced on several woody indicator species/cultivars are indicative of biological differences between the two inocula. Their nucleotide sequences are 92.2% identical and display between 79.2% and 97.1% identity with other ASGV isolates available in GenBank. The two isolates (M219-3 and M220) shared 92.3% deduced amino acid (daa) sequence identity for ORF1, 95.3% for ORF2 and 93.3% for the CP gene. A high degree of daa identity, 95.8% and 96.6%, was ob- served between the CPs of M219 ? 3 and M220, respectively, with the Brazilian isolate UV01. Potential recombination events and phy logenetic relationships to major parents were investigated in isolates M219 ? 3and M220. Keywords ASGV . Apple virus . Phylogenetic analysis . Recombinatio

Chiral Modulations in Curved Space I: Formalism

The goal of this paper is to present a formalism that allows to handle four-fermion effective theories at finite temperature and density in curved space. The formalism is based on the use of the effective action and zeta function regularization, supports the inclusion of inhomogeneous and anisotropic phases. One of the key points of the method is the use of a non-perturbative ansatz for the heat-kernel that returns the effective action in partially resummed form, providing a way to go beyond the approximations based on the Ginzburg-Landau expansion for the partition function. The effective action for the case of ultra-static Riemannian spacetimes with compact spatial section is discussed in general and a series representation, valid when the chemical potential satisfies a certain constraint, is derived. To see the formalism at work, we consider the case of static Einstein spaces at zero chemical potential. Although in this case we expect inhomogeneous phases to occur only as meta-stable states, the problem is complex enough and allows to illustrate how to implement numerical studies of inhomogeneous phases in curved space. Finally, we extend the formalism to include arbitrary chemical potentials and obtain the analytical continuation of the effective action in curved space.Comment: 22 pages, 3 figures; version to appear in JHE

Hall viscosity from gauge/gravity duality

In (2+1)-dimensional systems with broken parity, there exists yet another transport coefficient, appearing at the same order as the shear viscosity in the hydrodynamic derivative expansion. In condensed matter physics, it is referred to as "Hall viscosity". We consider a simple holographic realization of a (2+1)-dimensional isotropic fluid with broken spatial parity. Using techniques of fluid/gravity correspondence, we uncover that the holographic fluid possesses a nonzero Hall viscosity, whose value only depends on the near-horizon region of the background. We also write down a Kubo's formula for the Hall viscosity. We confirm our results by directly computing the Hall viscosity using the formula.Comment: 12 page

Analyzes of mealybug (Pseudococcus longispinus) virome reveal grapevine viruses diversity.

The long-tailed mealybug, Pseudococcus longispinus, is an important insect pest in grapevine growing areas in several countries, including Brazil. Metagenomic analysis of nucleic acids extracted from insect vectors makes it possible to study the diversity of insect viruses in addition to plant pathogenic viruses. In this study, insects (Ps. longispinus) were collected, and pooled throughout a plot of virus disease symptomatic vines, cultivated in growing beds, and analyzed by high throughput sequencing (HTS). The complete genome of grapevine leafroll-associated virus 2 and 3 (GLRaV-2 and -3) and a partial sequence of grapevine virus A (GVA) with two complete ORFs (coat protein and RNA-binding protein) were assembled from mealybug extracts and exhibited high nucleotide identities, up to 99%, with previously characterized homologous Brazilian isolates from grapevines. This information was validated by the detection of these viruses in the original symptomatic vines (N=76), from where mealybugs were collected, equivalent to an incidence of 34.2%, 89.5% and 36.8% for GLRaV-2, GLRaV-3 and GVA, respectively. Although one of these viruses is not transmitted by mealybugs (GLRaV-2), prospective of plant viruses infecting grapevine plants by analyzing the metagenome of insects could represent a relevant alternative to improve monitoring of viral diseases aiming at the management and control of viral diseases in vineyards or cultivation fields. This work is the first analysis of the Ps. longispinus virome in Brazil focusing on grapevine viruses