Molecular characterization of Prunus necrotic ringspot virus isolated from rose in Brazil / Caracterização molecular de Prunus necrotic ringspot vírus isolado de roseira no Brasil

There is no molecular characterization of Brazilian isolates of Prunus necrotic ringspot virus (PNRSV), except for those infecting peach. In this research, the causal agent of rose mosaic was determined and the movement (MP) and coat (CP) protein genes of a PNRSV isolate from rose were molecularly characterized for the first time in Brazil. The nucleotide and deduced amino acid sequences of MP and CP complete genes were aligned and compared with other isolates. Molecular analysis of the MP and CP nucleotide sequences of a Brazilian PNRSV isolate from rose and others from this same host showed highest identities of 96.7% and 98.6%, respectively, and Rose-Br isolate was classified in PV32 group.

Inexiste caracterização molecular de isolados brasileiros de Prunus necrotic ringspot virus (PNRSV), exceto aqueles de pessegueiros. Neste trabalho, o agente causal do mosaico da roseira foi determinado e os genes das proteínas de movimento (MP) e capsidial (CP) de um isolado de PNRSV de roseira foram molecularmente caracterizados pela primeira vez no Brasil. As sequências completas de nucleotídeos e de aminoácidos deduzidos da MP e da CP foram alinhadas e comparadas com outros isolados. Análises das sequências de nucleotídeos da MP e da CP do isolado brasileiro de PNRSV de roseira e outros isolados da mesma hospedeira revelaram altas identidades de 96,7% e 98,6%, respectivamente, sendo o isolado Rose-Br classificado no grupo PV32.

Molecular characterization of Prunus necrotic ringspot virus isolated from rose in Brazil / Caracterização molecular de Prunus necrotic ringspot vírus isolado de roseira no Brasil

There is no molecular characterization of Brazilian isolates of Prunus necrotic ringspot virus (PNRSV), except for those infecting peach. In this research, the causal agent of rose mosaic was determined and the movement (MP) and coat (CP) protein genes of a PNRSV isolate from rose were molecularly characterized for the first time in Brazil. The nucleotide and deduced amino acid sequences of MP and CP complete genes were aligned and compared with other isolates. Molecular analysis of the MP and CP nucleotide sequences of a Brazilian PNRSV isolate from rose and others from this same host showed highest identities of 96.7% and 98.6%, respectively, and Rose-Br isolate was classified in PV32 group.(AU)

Inexiste caracterização molecular de isolados brasileiros de Prunus necrotic ringspot virus (PNRSV), exceto aqueles de pessegueiros. Neste trabalho, o agente causal do mosaico da roseira foi determinado e os genes das proteínas de movimento (MP) e capsidial (CP) de um isolado de PNRSV de roseira foram molecularmente caracterizados pela primeira vez no Brasil. As sequências completas de nucleotídeos e de aminoácidos deduzidos da MP e da CP foram alinhadas e comparadas com outros isolados. Análises das sequências de nucleotídeos da MP e da CP do isolado brasileiro de PNRSV de roseira e outros isolados da mesma hospedeira revelaram altas identidades de 96,7% e 98,6%, respectivamente, sendo o isolado Rose-Br classificado no grupo PV32.(AU)

Simultaneous detection of four viruses affecting apple and pear by molecular hybridization using a polyprobe / Detecção simultânea de quatro vírus que infectam macieiras e pereiras por meio de hibridização molecular com uma polissonda

The viruses Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple mosaic virus (ApMV) are common in apples and pears and main targets of detection in propagation materials. This study aimed at demonstrating the usefulness of the hybridization method with a non-radioactive probe for simultaneous detection of these four viruses. The sensitivity of this method was sufficiently high enabling the detection of ASGV, ACLSV, ASPV and ApMV in total RNA extracted from infected samples. The probe specificity was confirmed by reaction with homologous viral cDNA, individually cloned for each virus.

Os vírus Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) e Apple mosaic virus (ApMV) são comuns em macieiras e pereiras e alvos prioritários da detecção em materiais propagativos. Este trabalho objetivou demonstrar a viabilidade do método de hibridização com uma sonda não-radioativa para a detecção simultânea desses quatro vírus. A sensibilidade deste teste foi suficientemente alta para permitir a detecção do ASGV, ACLSV, ASPV e ApMV em RNA total extraído de amostras infectadas. A especificidade da sonda foi confirmada pela reação com cDNA viral homólogo, individualmente clonado, para cada vírus.

Simultaneous detection of four viruses affecting apple and pear by molecular hybridization using a polyprobe / Detecção simultânea de quatro vírus que infectam macieiras e pereiras por meio de hibridização molecular com uma polissonda

The viruses Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple mosaic virus (ApMV) are common in apples and pears and main targets of detection in propagation materials. This study aimed at demonstrating the usefulness of the hybridization method with a non-radioactive probe for simultaneous detection of these four viruses. The sensitivity of this method was sufficiently high enabling the detection of ASGV, ACLSV, ASPV and ApMV in total RNA extracted from infected samples. The probe specificity was confirmed by reaction with homologous viral cDNA, individually cloned for each virus.(AU)

Os vírus Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) e Apple mosaic virus (ApMV) são comuns em macieiras e pereiras e alvos prioritários da detecção em materiais propagativos. Este trabalho objetivou demonstrar a viabilidade do método de hibridização com uma sonda não-radioativa para a detecção simultânea desses quatro vírus. A sensibilidade deste teste foi suficientemente alta para permitir a detecção do ASGV, ACLSV, ASPV e ApMV em RNA total extraído de amostras infectadas. A especificidade da sonda foi confirmada pela reação com cDNA viral homólogo, individualmente clonado, para cada vírus.(AU)

Molecular and phylogenetic characterization based on the complete genome of a virulent pathotype of Newcastle disease virus isolated in the 1970s in Brazil.

Newcastle disease (ND) is caused by the avian paramyxovirus type 1 (APMV-1) or Newcastle disease virus (NDV) that comprises a diverse group of viruses with a single-stranded, negative-sense RNA genome. ND is one of the most important diseases of chickens, because it severely affects poultry production worldwide. In the 1970s, outbreaks of virulent ND were recorded in Brazil, and the strain APMV-1/Chicken/Brazil/SJM/75 (SJM) of NDV was isolated. This strain was characterized as highly pathogenic for chickens but not pathogenic for other bird species. Here we present the complete genome of NDV strain SJM and investigate the phylogenetic relationships of this virus with other NDV strains in terms of genome and proteins composition, as well as characterizing its evolution process. The NDV strain SJM is categorized as a velogenic virus and the complete genome is 15,192 nucleotides in length, consisting of six genes in the order 3'-NP-P-M-F-HN-L-5'. The presence of the major pathogenic determinant of NDV strains ((112)R-R-Q-K-R↓F(117)) was identified in the Fusion protein of the NDV strain SJM. In addition, phylogenetic analysis classified the NDV strain SJM as a member of class II, genotype V, and indicates that this virus help us in the understanding of the evolutionary process of strains belonging to this genotype. This study contributes to the growing interest involving the characterization of NDV isolates to improve our current understanding about the epidemiology, surveillance and evolution of the pathogenic strains.

Detecção e caracterização molecular dos genes da proteína capsidial de ilarvírus e ampelovírus que infectam fruteiras temperadas / Detection and molecular characterization of ilarvirus and ampelovirus coat protein genes infecting temperate fruit trees

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).

Among the main pathogens infecting temperate fruit trees are Prune dwarf virus, Apple mosaic virus and Grapevine leafroll-associated virus 1. In this work the detection and molecular characterization of the coat protein genes of isolates from these viral species were carried out. Total RNA was extracted from peach, apple and grapevine leaves and RT-PCR reactions were performed using specific primers to each virus. The amplified cDNA fragments were cloned and sequenced. High identities were observed between coat protein nucleotide sequences of Brazilian isolates of PDV, ApMV and GLRaV-1 and isolates from other countries, independently from geographic origin and host. Coat protein molecular weights of these viruses were estimated by Western blot to be ca. 24kDa (PDV), 26kDa (ApMV) and 39kDa (GLRaV-1).

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

Among the main pathogens infecting temperate fruit trees are Prune dwarf virus, Apple mosaic virus and Grapevine leafroll-associated virus 1. In this work the detection and molecular characterization of the coat protein genes of isolates from these viral species were carried out. Total RNA was extracted from peach, apple and grapevine leaves and RT-PCR reactions were performed using specific primers to each virus. The amplified cDNA fragments were cloned and sequenced. High identities were observed between coat protein nucleotide sequences of Brazilian isolates of PDV, ApMV and GLRaV-1 and isolates from other countries, independently from geographic origin and host. Coat protein molecular weights of these viruses were estimated by Western blot to be ca. 24kDa (PDV), 26kDa (ApMV) and 39kDa (GLRaV-1).

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).

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

Among the main pathogens infecting temperate fruit trees are Prune dwarf virus, Apple mosaic virus and Grapevine leafroll-associated virus 1. In this work the detection and molecular characterization of the coat protein genes of isolates from these viral species were carried out. Total RNA was extracted from peach, apple and grapevine leaves and RT-PCR reactions were performed using specific primers to each virus. The amplified cDNA fragments were cloned and sequenced. High identities were observed between coat protein nucleotide sequences of Brazilian isolates of PDV, ApMV and GLRaV-1 and isolates from other countries, independently from geographic origin and host. Coat protein molecular weights of these viruses were estimated by Western blot to be ca. 24kDa (PDV), 26kDa (ApMV) and 39kDa (GLRaV-1).

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).