Detection of Virus and Viroid Pathogens in Plants

Narayanasamy, P.

doi:10.1007/978-90-481-9754-5_2

Cited by 5 publications

(11 citation statements)

References 473 publications

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“…Electron microscopy can be used to measure virus titers directly in plant cells, but this is laborious and time-consuming and is therefore unsuitable for rapid pathogen quantitation. Rather than counting virions, it is easier to determine the concentration of the viral capsid protein, which increases in proportion to the severity of infection and can be quantified using calibration-based immunological assays, such as the ELISA (57). Magnetic immunodetection combined with immunofiltration has numerous advantages over classic immunological assays, which remain the assays that are the most widely used for virus detection in plants.…”

Section: Discussionmentioning

confidence: 99%

Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

Rettcher

Jungk

Kühn

et al. 2015

Appl Environ Microbiol

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c Plant pathogens cause major economic losses in the agricultural industry because late detection delays the implementation of measures that can prevent their dissemination. Sensitive and robust procedures for the rapid detection of plant pathogens are therefore required to reduce yield losses and the use of expensive, environmentally damaging chemicals. Here we describe a simple and portable system for the rapid detection of viral pathogens in infected plants based on immunofiltration, subsequent magnetic detection, and the quantification of magnetically labeled virus particles. Grapevine fanleaf virus (GFLV) was chosen as a model pathogen. Monoclonal antibodies recognizing the GFLV capsid protein were immobilized onto immunofiltration columns, and the same antibodies were linked to magnetic nanoparticles. GFLV was quantified by immunofiltration with magnetic labeling in a double-antibody sandwich configuration. A magnetic frequency mixing technique, in which a two-frequency magnetic excitation field was used to induce a sum frequency signal in the resonant detection coil, corresponding to the virus concentration within the immunofiltration column, was used for high-sensitivity quantification. We were able to measure GFLV concentrations in the range of 6 ng/ml to 20 g/ml in less than 30 min. The magnetic immunoassay could also be adapted to detect other plant viruses, including Potato virus X and Tobacco mosaic virus, with detection limits of 2 to 60 ng/ml.

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Section: Discussionmentioning

confidence: 99%

Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

Rettcher

Jungk

Kühn

et al. 2015

Appl Environ Microbiol

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“…The traditional methods for viroid detection, such as biological indexing or bioassay, developed before the physical and chemical nature of viroids was known, have been employed for certification and quarantine programmes in order to detect viroids in vegetatively propagated planting material (Goss, ; Narayanasamy, ,b; Kovalskaya & Hammond, ). With the development of molecular biology, more precise, reliable and rapid techniques have been developed, such as polyacrylamide gel electrophoresis (PAGE) (Singh & Clark, ).…”

Section: Methods For Viroid Detectionmentioning

confidence: 99%

“…Direct application of sap extracts and partially purified samples allows rapid analysis but, on the other hand, it poses a problem with high backgrounds. Depending on the use, in certain cases nucleic acid isolation cannot be avoided (Hadidi et al ., ; Narayanasamy, ). Northern (RNA) and Southern (DNA) blot hybridization have been applied to distinguish among viroids with high sequence similarity and differences in size or conformation (Hadidi et al ., ).…”

Section: Methods For Viroid Detectionmentioning

confidence: 99%

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Diagnostic techniques for viroids

et al. 2016

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The fast growth of the human population forces us to produce more food, but higher crop production also leads to the fast spread of diseases. Plant pathology deploys a wide range of methods that do not provide an adequate solution to all disease losses. In the case of viroids, therapeutic means of control are not available; therefore control strategies are more focused on the development of reliable detection methods to quickly exclude the infected plant material. Although viroids are the smallest and simplest plant pathogens, their identification and detection is not straightforward. Each viroid–host combination is specific, and for reliable identification, all steps from sampling to final detection must be performed accurately. In this review, several methods for viroid detection in various host plants are discussed, including their advantages and disadvantages. Even though relatively new molecular methods enable fast and sensitive detection of viroids, a combination of different methods gives the most reliable identification. Techniques based on nucleic acids may be the future for viroid detection but they still cannot replace biological indexing, which is usually essential in epidemiological and aetiological studies.

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“…Since the viruses may be deactivated quickly when the infected plants dry up under field conditions, few studies have been done to detect the Table 1 plant viruses in soil and water (Boben et al 2007;Yang et al 2012). Viroids represent a group of extremely primitive pathogenic entities consisting exclusively of nucleic acids that are capable of independent replication and inducing diseases when introduced into susceptible plant cells (Narayanasamy 2011). Because of the absence of a protein component that is present in the viruses, diagnostic approaches based upon serology have not been applicable for the detection of viroids.…”

Section: Detection Of Plant Pathogensmentioning

confidence: 99%

Real-time PCR applied to study on plant pathogens: potential applications in diagnosis - a review

Mirmajlessi¹,

Loit²,

Mänd³

et al. 2015

Plant Prot. Sci.

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Mirmajlessi S.M., Loit E., Mänd M., Mansouripour S.M. (2015): Real-time PCR applied to study on plant pathogens: potential applications in diagnosis -a review. Plant Protect Sci., 51: 177-190.Quantitative real-time PCR (qPCR) technique incorporates traditional polymerase chain reaction (PCR) efficiency with the production of a specific fluorescent signal, measuring the kinetics of the reaction in the early PCR phases and providing quantification of specific targets in various environmental samples. There are an increasing number of chemistries to detect PCR products, which are widely used in plant pathology as they cluster into the amplicon sequence non-specific and sequence-specific techniques. In this review, we illustrate a general description of major chemistries and discuss some considerations for assay development as it applies for a wide range of applications in epidemiological studies. The technique has become the gold standard for early detection of pathogens and a fundamental tool in the research laboratory.

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Detection of Virus and Viroid Pathogens in Plants

Cited by 5 publications

References 473 publications

Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

Simple and Portable Magnetic Immunoassay for Rapid Detection and Sensitive Quantification of Plant Viruses

Diagnostic techniques for viroids

Real-time PCR applied to study on plant pathogens: potential applications in diagnosis - a review

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