Sensitive Detection of Apple Chlorotic Leaf Spot Virus From Infected Apple or Peach Tissue Using Rt-Pcr, Ic-Rt-Pcr, or Multiplex Ic-Rt-PCR

Nemchinov, Lev G.; Hadidi, A.; Foster, Joseph; Candresse, Thierry; Verderevskaya, T. D.

doi:10.17660/actahortic.1995.386.4

Cited by 30 publications

(12 citation statements)

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“…The presence of genetically divergent ACLSV isolates implies several ACLSV introduction sources in Latvia, resulting in diversification and mixing of populations in the hosts by the propagation material (rootstocks and grafts). The great heterogeneity of the ACLSV population in general was considered as a reason for the absence of biological grouping of isolates in phylogenetic analyses based on their host species (Nemchinov et al. 1995).…”

Section: Discussionmentioning

confidence: 99%

Occurrence and Diversity of Pome Fruit Viruses in Apple and Pear Orchards in Latvia

Pūpola

Moročko‐Bičevska

Kāle

et al. 2011

Journal of Phytopathology

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Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV) and Apple mosaic virus are economically important viruses infecting fruit tree species worldwide. To evaluate the occurrence of these pome fruit viruses in Latvia, a large-scale survey was carried out in 2007. Collected samples were tested for infection by DAS ELISA and multiplex RT-PCR. The accuracy of the detection of the viruses in multiplex RT-PCR was confirmed by sequencing amplified PCR fragments. The results showed a wide occurrence of viruses in apple and pear commercial orchards established from nontested planting material. More than 89% of the tested apple trees and more than 60% of pear trees were infected with one or more pome fruit viruses. Analyses showed that the high occurrence of viruses in several apple cultivars is due to the propagation of infected clonal rootstocks and scions from infected mother trees. Sequence analyses targeting the 3¢-terminal region of the tested viruses showed various degrees of genetic diversity within respective virus isolates. This is the first report of the occurrence of ACLSV, ASGV and ASPV in apple and pear trees in Latvia and demonstrates their genetic diversity in different host genotypes.

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Section: Discussionmentioning

confidence: 99%

Occurrence and Diversity of Pome Fruit Viruses in Apple and Pear Orchards in Latvia

Pūpola

Moročko‐Bičevska

Kāle

et al. 2011

Journal of Phytopathology

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“…Multiplex PCR is useful in plant pathology because different bacteria and/or RNA viruses frequently infect a single host and consequently sensitive detection is needed for the propagation of pathogen-free plant material. There are several examples in plant pathology of simultaneous detection of several targets and the amplification by multiplex PCR of two or three plant viruses has been reported [22,24,38,40,56,57,60,61]. Nevertheless, there are still only a few examples in which more than three plant viruses were amplified in a single PCR-based assay [4,39,41,44], probably due to the technical difficulties of designing a reaction involving many compatible primers.…”

Section: Multiplex Pcrmentioning

confidence: 99%

Innovative tools for detection of plant pathogenic viruses and bacteria

et al. 2003

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Detection of harmful viruses and bacteria in plant material, vectors or natural reservoirs is essential to ensure safe and sustainable agriculture. The techniques available have evolved significantly in the last few years to achieve rapid and reliable detection of pathogens, extraction of the target from the sample being important for optimising detection. For viruses, sample preparation has been simplified by imprinting or squashing plant material or insect vectors onto membranes. To improve the sensitivity of techniques for bacterial detection, a prior enrichment step in liquid or solid medium is advised. Serological and molecular techniques are currently the most appropriate when high numbers of samples need to be analysed. Specific monoclonal and/or recombinant antibodies are available for many plant pathogens and have contributed to the specificity of serological detection. Molecular detection can be optimised through the automatic purification of nucleic acids from pathogens by columns or robotics. New variants of PCR, such as simple or multiplex nested PCR in a single closed tube, co-operative-PCR and real-time monitoring of amplicons or quantitative PCR, allow high sensitivity in the detection of one or several pathogens in a single assay. The latest development in the analysis of nucleic acids is micro-array technology, but it requires generic DNA/RNA extraction and pre-amplification methods to increase detection sensitivity. The advances in research that will result from the sequencing of many plant pathogen genomes, especially now in the era of proteomics, represent a new source of information for the future development of sensitive and specific detection techniques for these microorganisms.

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“…Multiplex reverse transcriptase PCR (mRT‐PCR) has been used to simultaneously amplify more than one locus in the same reaction, proving to be a rapid, cost‐effective screening assay for plant viruses (Bariana et al ; Nemchinov et al ; Henegariu et al ; Matic et al ; Noorani et al ; Ge et al ). Various types of mRT‐PCR assays have been previously applied to detect viruses that affect stone fruit trees.…”

Section: Introductionmentioning

confidence: 99%

Multiplex RT‐PCR to simultaneously detect three viruses that infect peach

Yang

Zhou

et al. 2019

Lett Appl Microbiol

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Significance and Impact of the Study: In this study, we developed a multiplex reverse transcriptase PCR (mRT-PCR) assay for simultaneously detecting three viruses that infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). This assay is simple, easy to perform, reliable and cost-effective, and can thus be applied for large-scale surveys of PaLV, PeVD and NSPaV. AbstractPeach is a major crop in China, and like any other stone fruit, virus and viruslike diseases can reduce the yield and quality of the fruit. Herein, we developed a multiplex RT-PCR (mRT-PCR) assay for simultaneously detecting three viruses known to infect peach: peach-associated luteovirus (PaLV), peach virus D (PeVD) and nectarine stem-pitting-associated virus (NSPaV). Plant nad5 mRNA was used as the internal control. Field samples that were co-infected with PaLV, PeVD and NSPaV were used; we identified three primer pairs to be the most specific for detecting these viruses, followed by determining the ideal concentration of each primer pair and optimizing the annealing temperature for mRT-PCR. We also assessed the detection limit using serial dilutions of RNA and cDNA. The newly developed mRT-PCR assay could simultaneously detect PaLV, PeVD and NSPaV. To validate the reliability of mRT-PCR for virus detection, mRT-PCR was used to detect viruses in the leaves of 21 peach plants collected in Liaoning Province, China. The obtained results revealed the presence of single and co-infections. To conclude, the mRT-PCR assay developed herein is sensitive, reliable and economical, and we believe that it can thus be used for large-scale surveys of PaLV, PeVD and NSPaV.Letters in Applied Microbiology ISSN 0266-8254

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Sensitive Detection of Apple Chlorotic Leaf Spot Virus From Infected Apple or Peach Tissue Using Rt-Pcr, Ic-Rt-Pcr, or Multiplex Ic-Rt-PCR

Cited by 30 publications

References 0 publications

Occurrence and Diversity of Pome Fruit Viruses in Apple and Pear Orchards in Latvia

Occurrence and Diversity of Pome Fruit Viruses in Apple and Pear Orchards in Latvia

Innovative tools for detection of plant pathogenic viruses and bacteria

Multiplex RT‐PCR to simultaneously detect three viruses that infect peach

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