Real-time PCR detection and quantification of vector trichodorid nematodes and Tobacco rattle virus

Holeva, R. C.; Phillips, M. S.; Neilson, Roy; Brown, D. J. F.; Young, Virginia A.; Boutsika, Konstantina; Blok, Vivian C.

doi:10.1016/j.mcp.2005.12.004

Cited by 35 publications

(14 citation statements)

References 24 publications

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“…Using sequence data, Boutsika et al (2004a) and Riga et al (2007) developed a PCR with specific primers for diagnostics of P. allius, P. macrostylus, P. pachydermus, P. teres, T. primitivus and T. similis. Holeva et al (2006) designed a real-time PCR assay for detection and quantification of P. pachydermus and T. similis in field samples. Recently, Duarte et al (2011) developed a PCR-RFLP assay based on the 18S rRNA gene for rapid identification of 12 trichodorid species belonging to the genera Nanidorus, Paratrichodorus and Trichodorus.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Kumari

Subbotin

2012

Plant Pathology

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Plant parasitic nematodes of the family Trichodoridae cause substantial yield losses in many agricultural crops. Rapid and accurate identification of trichodorids to the species level is critical for selection of appropriate measures for control. This study analysed 99 sequences of the D2-D3 expansion segments of the 28S rRNA gene and 131 sequences of the 18S rRNA gene from the stubby nematodes belonging to the genera Nanidorus, Paratrichodorus and Trichodorus. Species delimiting was based on the integration of morphological identification, which is not provided in the present article, and molecularbased phylogenetic inference and sequence analysis. Twenty-two valid species and several species complexes were identified among nematodes included in the analysis. PCR-RFLPs of the partial 18S rDNA and the D2-D3 expansion segments of the 28S rDNA were tested and proposed for identification of these nematodes. Gel PCR-RFLP profiles and tables with restriction fragment lengths for several diagnostic enzymes are provided for identification. Some problems of taxonomy and phylogeny of nematodes of the family Trichodoridae are also discussed.

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Section: Introductionmentioning

confidence: 99%

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Kumari

Subbotin

2012

Plant Pathology

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“…Coupled to automated product analysis software, the real-time PCR technique allows continuous monitoring of the sample during PCR using hybridization probes or dyes (Kubista et al 2006). Real-time PCR techniques have been developed to detect plant-parasitic nematodes, including PWN Holeva et al 2006;François et al 2007;Leal et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Detection of the pine wood nematode using a real-time PCR assay to target the DNA topoisomerase I gene

Huang

et al. 2010

Eur J Plant Pathol

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A species-specific real-time PCR assay targeting the DNA topoisomerase I gene has been developed to detect Bursaphelenchus xylophilus. The specificity of the assay was confirmed by the lack of amplification of genomic DNA from other Bursaphelenchus or Seinura species. The sensitivity test showed that the limit of the reaction was 0.01 ng of genomic DNA or one individual nematode, as small as an egg. The validity of the real-time PCR assay was evaluated by analyzing mixed nematode samples extracted from diseased pine trees in which B. xylophilus was associated with several closely related species, B. mucronatus, B. hofmanni, Aphelenchoides macronucleatus, S. lii and S. wuae. These results demonstrate the potential of the assay to provide rapid, specific and sensitive molecular identification of B. xylophilus for use in pest risk assessment and quarantine regulations.

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“…By employing the protocol developed in this investigation, both vector species and TRV RNA-1 and RNA-2 could be detected in the field soil samples. This assay has the potential to provide rapid, accurate and sensitive molecular information for risk assessment in the field (Holeva et al 2006).…”

Section: Nucleic Acid-based Techniquesmentioning

confidence: 99%

Detection of Viral and Viroid Plant Pathogens in the Environment

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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Plant viruses and viroids with obligate parasitism as the mode of existence, have to be in association with living cells of plant hosts always. However, some highly stable viruses have been detected in a free state in soil, water or air. Other viruses depend on the vectors such as insects, mites, nematodes or fungi present in the environment for their spread from infected plants to healthy plants which may be available in the same field or far away. The role of alternative/additional host plant species functioning as sources of virus infection has been demonstrated by applying various diagnostic techniques. The presence of viruses has been detected by employing biological, immunological and nucleic acid-based techniques. The effectiveness and reliability of detection varies significantly. However, molecular techniques have been shown to be more sensitive, specific and fast in providing the results, although they are unable to indicate the level of pathogenic potential of the viruses detected in the environmental samples. Viroids lacking natural vectors and stability outside living cells have been detected in several additional/alternative plant species which may function as reservoir of viroid infection.

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Real-time PCR detection and quantification of vector trichodorid nematodes and Tobacco rattle virus

Cited by 35 publications

References 24 publications

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Molecular characterization and diagnostics of stubby root and virus vector nematodes of the family Trichodoridae (Nematoda: Triplonchida) using ribosomal RNA genes

Detection of the pine wood nematode using a real-time PCR assay to target the DNA topoisomerase I gene

Detection of Viral and Viroid Plant Pathogens in the Environment

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