Serological and Molecular Detection of Latent Viruses in the Apple Germplasm Bank of Santa Catarina

Abstract The Apple Germplasm Bank (AGB) of Santa Catarina Agricultural Research and Rural Extension Company - Epagri, AGB-Epagri, is the largest of the genus Malus in Brazil. Twenty-eight main accessions of this bank were virus screened through DAS-ELISA, RT-PCR and IC-RT-PCR during two consecutive reproductive cycles, and each accession showed latent mixed infection by at least two species, among ASGV, ASPV and ACLSV. The combined use of diagnostic methods helped overcome inconsistencies commonly found in apple virus detection and was shown essential for the AGB-Epagri can be safely used as a source of genetic variability and for the exchange of virus-free propagative material.

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)