Improvement of Potato virus Y (PVY) detection and quantitation using PVYN- and PVYO-specific real-time RT-PCR assays

Balme-Sinibaldi, Valérie; Tribodet, Michel; Croizat, Flora; Lefeuvre, Pierre; Kerlan, Camille; Jacquot, Emmanuel

doi:10.1016/j.jviromet.2006.01.019

Cited by 39 publications

(21 citation statements)

References 58 publications

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“…Isolates of the PVY C strain however cause systemic necrosis, a symptom known as stipple streak in potato cultivars carrying the Nc gene (De Bokx and Huttinga 1981). The different strains cannot be distinguished in the field, but by laboratory techniques as the serological method DAS-ELISA, using monoclonal antibodies, or the RT-PCR technique, or by use of indicator plants, a differentiation is perfectly possible (Balme-Sinibaldi et al 2006, Glais et al 2002, Blanco-Urgoiti et al 1998.…”

Section: Introductionmentioning

confidence: 99%

Potato clones with multiple copies of the Ryadg allele conferring resistance to PVY

Andrade¹,

Pinto²,

Ribeiro³

et al. 2009

CBAB

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Section: Introductionmentioning

confidence: 99%

Potato clones with multiple copies of the Ryadg allele conferring resistance to PVY

Andrade¹,

Pinto²,

Ribeiro³

et al. 2009

CBAB

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“…(), only one (Balme‐Sinibaldi et al . ) reported the development of a real‐time PCR assay allowing the quantification during competition at the intraspecific level (PVY O and PVY N isolates of the potato virus Y). To our knowledge, all others methods consisted in the detection and quantification of pathogens at the intragenus (or even upper) level.…”

Section: Resultsmentioning

confidence: 99%

Specific detection and quantification of virulent/avirulent Phytophthora infestans isolates using a real-time PCR assay that targets polymorphisms of the Avr3a gene

Clément

Baldwin

Magalon

et al. 2013

Lett Appl Microbiol

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Significance and Impact of the Study: This tool clearly improves previously published methods of specific real-time quantitative PCR for Phytophthora infestans by allowing the intraspecific detection and quantification of Avr3a/avr3a genotypes. This reliable and sensitive assay has enabled to assess zoospore production as a fitness proxy and thus offers new opportunities for future researches on P. infestans/ host quantitative interactions. Abstract Molecular tools that allow intraspecific quantification and discrimination of pathogen isolates are useful to assess fitness of competitors during mixed infections. However, methods that were developed for quantifying Phytophthora infestans are only specific at the species level. Here, we reported a TaqMan-based real-time PCR assay allowing, according to the specificity of the used probes, an accurate quantification of different proportions of two genetically distinct clones of P. infestans in mixed fractions. Indeed, in addition to a primer specific to P. infestans, two primers and two TaqMan â probes that target single-nucleotide polymorphisms located in the Avr3a/avr3a virulence gene sequence were designed. The reliability of the method was tested on serially diluted fractions containing plasmid DNA with either the Avr3a or the avr3a sequences at concentrations ranging from 10 2 to 10 8 copies per ll. Based on its specificity, sensitivity and repeatability, the proposed assay allowed a quantification of the targeted DNA sequence in fractions with a Avr3a/avr3a ratio in the range 1/99 to 99/1. The reliability of the test was also checked for counting zoospores. Applications for future research in P. infestans/host quantitative interactions were also discussed.

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“…Further study on PVY/potato interactions should now be carried out to identify the plant product(s) interacting with the E419 of the HC-Pro protein. -Sinibaldi et al (2006), Glais and Jacquot (2015), Jacquot et al (2005), Kerlan et al (1999) and Rolland et al (2008).…”

Section: Discussionmentioning

confidence: 99%

The amino acid 419 in HC-Pro is involved in the ability of PVY isolate N605 to induce necrotic symptoms on potato tubers

et al. 2015

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The ability to induce the potato tuber necrosis ringspot disease (PTNRD) is a property shared by PVY isolates belonging to different groups (e.g. PVY(N) and PVY(O)) and variants (e.g. PVY(NTN) and PVY(N)-W). The identification of viral molecular determinant(s) involved in the expression of PTNRD symptoms is essential for (i) an easier detection of tuber necrosis isolates and (ii) an improvement of our knowledge on the epidemiology of this potato disease. A reverse genetic approach associated with a biological typing of a collection of PVY chimeras and mutants indicated that residue E419 of the HC-Pro protein is linked to the ability of PVY to induce tuber necrosis on four PTNRD-susceptible potato cultivars. Indeed, the substitution of the N-type glutamic acid (E) in O-type aspartic acid (D) at position 419 in the HC-Pro cistron prevents the expression of tuber necrosis on infected tubers without reducing the virulence of the corresponding E/D419 mutant. This result opens opportunities for the future studies on potato/PVY interactions.

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Improvement of Potato virus Y (PVY) detection and quantitation using PVYN- and PVYO-specific real-time RT-PCR assays

Cited by 39 publications

References 58 publications

Potato clones with multiple copies of the Ryadg allele conferring resistance to PVY

Potato clones with multiple copies of the Ryadg allele conferring resistance to PVY

Specific detection and quantification of virulent/avirulent Phytophthora infestans isolates using a real-time PCR assay that targets polymorphisms of the Avr3a gene

The amino acid 419 in HC-Pro is involved in the ability of PVY isolate N605 to induce necrotic symptoms on potato tubers

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