Obtenção de material propagativo livre de vírus e diagnóstico de vírus em macieiras e pereiras.

Principais vírus da macieira e da pereira. Danos causados por vírus. Limpeza clonal: produção de macieiras e pereiras livres de vírus. Análises e testes de avaliação de sanidade.bitstream/item/60758/1/CNPUV-DOC.-69-09.pd

Detection of Viruses in Apples and Pears by Real Time RT-PCR Using 5'-Hydrolysis Probes

Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV) and Apple mosaic virus (ApMV) are common in apples and pears and the main targets of virus elimination from propagation material. The objective of this work was to design primers and probes for a real time RT-PCR protocol for detection of the four above viruses. FAM/TAMRA-labeled probes and primers were designed by searching for highly conserved nucleotide regions in the coat protein gene of the four viruses. Infection levels in analyzed apple samples were 92.6, 96.4, 100 and 88% for ASGV, ASPV, ACLSV and ApMV, respectively. In pears, all pre-existing ASPV infections were detected. Viral infections were confirmed in a selection of commercial cultivars of apples and pear scions, and quince rootstocks, demonstrating the sensitivity and reliability of the designed primers and probes. Real time RT-PCR using 5'-labeled probes is suitable for checking sanitary quality as a routine test in certification programs

First report on detection of three Bunya-Like Viruses in apples in Brazil.

Apple chlorotic leaf spot virus (ACLSV), apple stem grooving virus (ASGV), and apple stem pitting virus (ASPV) cause significant losses to Brazilian (BR) apple production. Looking beyond these latent viruses, high-throughput sequencing (HTS) of three samples (Vacaria, Brazil) was performed on an Illumina HiSeq X Ten platform (USA), cv. Braeburn (BB), and a BGISEQ-500 platform (China), cvs. Royal Gala (RG) and Mishima (MI)

Detecção e caracterização molecular dos genes da proteína capsidial de ilarvírus e ampelovírus que infectam fruteiras temperadas.

Dentre os principais patógenos que incidem em fruteiras temperadas, destacam-se o Prune dwarf virus (PDV), o Apple mosaic virus (ApMV) e o Grapevine leafroll-associated virus 1 (GLRaV-1). Neste trabalho foram realizadas a detecção e a caracterização molecular dos genes da proteína capsidial de isolados destas três espécies virais. RNAs totais foram extraídos de amostras de folhas de pessegueiros, macieiras e videiras e, nas reações de RT-PCR, foram utilizados oligonucleotídeos específicos para cada espécie viral. Os cDNAs amplificados foram clonados e sequenciados. Foram verificadas altas identidades entre as sequências de nucleotídeos dos genes da proteína capsidial dos isolados brasileiros de PDV, ApMV e GLRaV-1 e isolados de outros países, independente da origem geográfica e da hospedeira. O peso molecular da proteína capsidial destes vírus foi estimado por meio de Western blot em cerca de 24kDa (PDV), 26kDa (ApMV) e 39kDa (GLRaV-1).Nota técnica

Functional Relaxation and Guided Imagery as Complementary Therapy in Asthma: A Randomized Controlled Clinical Trial

Background: Asthma is a frequently disabling and almost invariably distressing disease that has a high overall prevalence. Although relaxation techniques and hypnotherapeutic interventions have proven their effectiveness in numerous trials, relaxation therapies are still not recommended in treatment guidelines due to a lack of methodological quality in many of the trials. Therefore, this study aims to investigate the efficacy of the brief relaxation technique of functional relaxation (FR) and guided imagery (GI) in adult asthmatics in a randomized controlled trial. Methods: 64 patients with extrinsic bronchial asthma were treated over a 4-week period and assessed at baseline, after treatment and after 4 months, for follow-up. 16 patients completed FR, 14 GI, 15 both FR and GI (FR/GI) and 13 received a placebo relaxation technique as the control intervention (CI). The forced expiratory volume in the first second (FEV 1) as well as the specific airway resistance (sR(aw)) were employed as primary outcome measures. Results: Participation in FR, GI and FR/GI led to increases in FEV 1 (% predicted) of 7.6 +/- 13.2, 3.3 +/- 9.8, and 8.3 +/- 21.0, respectively, as compared to -1.8 +/- 11.1 in the CI group at the end of the therapy. After follow-up, the increases in FEV 1 were 6.9 +/- 10.3 in the FR group, 4.4 +/- 7.3 in the GI and 4.5 +/- 8.1 in the FR/GI, compared to -2.8 +/- 9.2 in the CI. Improvements in sR(aw) (% predicted) were in keeping with the changes in FEV 1 in all groups. Conclusions: Our study confirms a positive effect of FR on respiratory parameters and suggests a clinically relevant long-term benefit from FR as a nonpharmacological and complementary therapy treatment option. Copyright (C) 2009 S. Karger AG, Base

Detection and molecular characterization of Grapevine yellow speckle viroid 1 isolates infecting grapevines in Brazil.

Presently, Hop stunt viroid(HSVd) and Citrus exocortis viroid (CEVd) are the only viroids reported to infect grapevines (Vitis spp.) in Brazil, among the seven viroid species already reported infecting this host in other countries. All grapevine viroid diseases are graft-transmissible and can induce losses especially when associated with viruses. The aim of this work was to confirm infection by Grapevine yellow speckle viroid 1(GYSVd-1) in grapevine samples exhibiting yellow speckle symptoms in the leaves and in asymptomatic samples sequenced by next generation sequencing (NGS). The occurrence of this viroid in Brazil was further investigated in a second study. Total RNAs and dsRNAs were extracted from five symptomatic plants and 16 asymptomatic samples, respectively. Specific primers were used for RT-PCR and amplified DNA fragments were cloned and sequenced by the Sanger method. Eleven complete nucleotide sequences of GYSVd-1 isolates (366 ?367 nt) were obtained from NGS and from RT-PCR amplicons. Comparisons showed high identities (95.9 ?100 %) among ten isolates and an identity of 87.2 ?90.4 % with a divergent isolate (RM-BR). Phylogenetic analyses placed GYSVd-1 isolates in four clusters (types 1, 2, 3 and 4). All GYSVd-1 infections were confirmed by conventional RT-PCR and RT-qPCR using specific oligonucleo-tides and a labeled probe. This is the first report and molecular characterization of GYSVd-1 infecting grapevines in Brazil, and our survey indicates that this viroid could be widespread in the major grape producing regions of Brazil. Keywords GYSVd-1 . Incidence . Next generation sequencing. Secondary structure. Vine

On equivariant characteristic ideals of real classes

Let $p$ be an odd prime, $F/{\Bbb Q}$ an abelian totally real number field, $F_\infty/F$ its cyclotomic ${\Bbb Z}_p$-extension, $G_\infty = Gal (F_\infty / {\Bbb Q}),$ ${\Bbb A} = {\Bbb Z}_p [[G_\infty]].$ We give an explicit description of the equivariant characteristic ideal of $H^2_{Iw} (F_\infty, {\Bbb Z}_p(m))$ over ${\Bbb A}$ for all odd $m \in {\Bbb Z}$ by applying M. Witte's formulation of an equivariant main conjecture (or "limit theorem") due to Burns and Greither. This could shed some light on Greenberg's conjecture on the vanishing of the $\lambda$-invariant of $F_\infty/F.

First report of grapevine pinot gris virus infecting grapevine in Brazil.

Grapevine Pinot gris virus (GPGV) has been re-ported infecting grapevines (Vitisspp.) in several countriesof the world. In this study, 19.5% of grapevine samples col-lected in Brazil and indexed for GPGV by RT-qPCR wasinfected. The complete coat protein and the partial replicase genes of two Brazilian isolates (CF-BR and ME-BR) were sequenced and exhibited very high nucleotide and deduced amino acid identities with many foreign characterized isolates of GPGVavailable in the GenBank. This is the first detection of GPGV infecting grapevines in Brazil. Keywords GPGV . Trichovirus . RT-qPCR . Real time RT-PCR. Diagnosis.Disponível em: . Acesso em: 04 set. 2017

Absolute quantification of viruses by TaqMan real-time RT-PCR in grapevines.

The absolute quantification determines the absolute amount of a targeted nucleic acid expressed as a copy number or concentration. The knowledge of virus concentrations in commercial crops possesses high relevance to ensure a reliable diagnosis. The objective of this study was to perform an absolute quantification of five viruses in infected grapevines (Vitis spp.). Different known amounts of the standard sample (cloned viral cDNA or in vitro transcribed viral RNA) were quantified by TaqMan RT-qPCR. Based on these data, standard curves were generated plotting Ct values (threshold cycle) against the log of the standard sample amount. Infected grapevine samples were evaluated to determine virus titers, which were highly variable. This result may contribute to improve virus diagnosis by accurately quantifying virus titre variations in grapevines. Key words: RT-qPCR, GRSPaV, GVA, GVD, GLRaV-3 and -4. RESUMO: A quantificação absoluta determina a quantidade absoluta de um ácido nucleico alvo expressa como número de cópias ou concentração. O conhecimento das concentrações virais em culturas comerciais tem grande relevância para assegurar um diagnóstico confiável. O objetivo deste trabalho foi realizar uma quantificação absoluta de cinco vírus em videiras infectadas (Vitis spp.). Diferentes quantidades conhecidas da amostra padrão (cDNA viral clonado ou RNA viral transcrito in vitro) foram quantificadas por RT-qPCR TaqMan. A partir destes dados, curvas padrão foram geradas plotando-se os valores de Ct (ciclo limiar) contra o log da quantidade da amostra padrão. Amostras de videiras infectadas foram avaliadas visando-se determinar os títulos virais que foram bastante variáveis. Este resultado contribui para melhorar o diagnóstico viral ao quantificar com precisão variações no título viral em videiras. Palavras-chave: RT-qPCR, GRSPaV, GVA, GVD, GLRaV-3 e -4. A quantificação absoluta determina a quantidade absoluta de um ácido nucleico alvo expressa como número de cópias ou concentração. O conhecimento das concentrações virais em culturas comerciais tem grande relevância para assegurar um diagnóstico confiável. O objetivo deste trabalho foi realizar uma quantificação absoluta de cinco vírus em videiras infectadas (Vitis spp.). Diferentes quantidades conhecidas da amostra padrão (cDNA viral clonado ou RNA viral transcrito in vitro) foram quantificadas por RT-qPCR TaqMan. A partir destes dados, curvas padrão foram geradas plotando-se os valores de Ct (ciclo limiar) contra o log da quantidade da amostra padrão. Amostras de videiras infectadas foram avaliadas visando-se determinar os títulos virais que foram bastante variáveis. Este resultado contribui para melhorar o diagnóstico viral ao quantificar com precisão variações no título viral em videiras. Palavras-chave: RT-qPCR, GRSPaV, GVA, GVD, GLRaV-3 e -4