Molecular characterization of potato virus YN isolates by PCR-RFLP

Glais, Laurent; Kerlan, Camille; Tribodet, Michel; Tordo, V. Marie Jeanne; Robaglia, Christophe; Astier-Manifacier, Suzanne

doi:10.1007/bf01877246

Cited by 59 publications

(25 citation statements)

References 28 publications

(32 reference statements)

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“…To corroborate our results, we compared the findings of the Nco I/Bgl II method with those of the RFLP analysis of Glais et al (1996). Of the three enzymes that were used by those authors we found Hinc II to be the most reliable: although it cleaved the PCR products of both N and NTN strains, the numbers of cleavage sites differed between the strains: one in the NTN and two in the N (see Figs 1 and 4).…”

Section: Test For Pvy Nlw In Field Samplessupporting

confidence: 81%

“…Some of the enzymes which were previously used (Glais et al, 1996) differentiated between the N and NTN strains but they cleave at several sites within the virus amplicons of both the NTN and N strains (e.g., Taq I and Hinc II). Others (Ava II) cleave similarly in the amplicons of both strains (see Fig.…”

Section: Discussionmentioning

confidence: 99%

“…PCR primers were derived from the 5' end of the PVY genome; primers c (forward) and d (reverse) were as in Glais et al (1996). The PCR reaction mixture and conditions were as described in Rosner et al (1998).…”

Section: Virus Isolatesmentioning

confidence: 99%

“…Differentiation of PVY NTN from PVY N by RFLP of PCR products amplified from the 5' end region of the virus genome was demonstrated by Glais et al (1996). A more comprehensive restriction map was constructed by amplifying the whole PVY genome in an attempt to locate specific gene functions (Glais et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Differentiating PVYNTN by unique single-restriction cleavage of PCR products

Rosner

Maslenin

1999

Potato Res

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SummaryA procedure for differentiating PVY NTN from PVY TM is described and is based on the unique cleavage of their respective PCR products with strain specific restriction endonucleases. The PCR products corresponding to the 5' end of the N and NTN strains of PVY were cloned and sequenced, and a restriction map was constructed which included common enzymes that were used for the differentiation of PVY NTN. Unique. single cleavage of PCR products derived from the 5' end of the PVY NTN genome by Nco I, and that of the N-strain of PVY by Bgl II restriction endonuclease were demonstrated. The specific digestion patterns in polyacrylamide gel were used for the unequivocal differentiation between the N and NTN strains of the virus. Both single and mixed infections were detected in field samples of potatoes using this procedure.

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Section: Test For Pvy Nlw In Field Samplessupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Virus Isolatesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Differentiating PVYNTN by unique single-restriction cleavage of PCR products

Rosner

Maslenin

1999

Potato Res

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“…The symptomology results described above showed that typical pepper-PVY isolates were distinguished from potato isolates by host range (8). After phylogenetic analysis we noticed that pepper isolate has high similarity with LYE84.2 strain that was isolated from tomato (6) and with SON41 strain witch was isolated from C. annuum (5). For this reason pepper infecting isolates could be classified within the PVY NP group that expanding earlier suggestions made by Romero et al (10).…”

Section: Discussionsupporting

confidence: 67%

Study of biological and molecular characterization of pepper-PVY isolated from Tehran pepper fields and it’s comparison with other PVY isolates

et al. 2007

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Abstract:Potato virus Y a type species of the genus potyvirus infects several crops in the family solanaceae. PVY isolated from field infected peppers was identified on the basis of host reaction, serological and molecular characterization. The result of ELISA, Immunoblot electrophoresis and Immuno Capture Reverse Transcription Polymerase Chain Reaction (IC-RT-PCR) indicated that pepper isolate of PVY shares the reported properties of the PVY but in host range studies some differences were observed. For this reason the sequence analysis was performed .This is the first report of PVY incidence in Iran pepper fields.

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Detection and Characterization of Viral Species/Subspecies Using Isothermal Recombinase Polymerase Amplification (RPA) Assays

Glais

Jacquot

2015

Plant Pathology

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Numerous molecular-based detection protocols include an amplification step of the targeted nucleic acids. This step is important to reach the expected sensitive detection of pathogens in diagnostic procedures. Amplifications of nucleic acid sequences are generally performed, in the presence of appropriate primers, using thermocyclers. However, the time requested to amplify molecular targets and the cost of the thermocycler machines could impair the use of these methods in routine diagnostics. Recombinase polymerase amplification (RPA) technique allows rapid (short-term incubation of sample and primers in an enzymatic mixture) and simple (isothermal) amplification of molecular targets. RPA protocol requires only basic molecular steps such as extraction procedures and agarose gel electrophoresis. Thus, RPA can be considered as an interesting alternative to standard molecular-based diagnostic tools. In this paper, the complete procedures to set up an RPA assay, applied to detection of RNA (Potato virus Y, Potyvirus) and DNA (Wheat dwarf virus, Mastrevirus) viruses, are described. The proposed procedure allows developing species- or subspecies-specific detection assay.

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Molecular characterization of potato virus YN isolates by PCR-RFLP

Cited by 59 publications

References 28 publications

Differentiating PVYNTN by unique single-restriction cleavage of PCR products

Differentiating PVYNTN by unique single-restriction cleavage of PCR products

Study of biological and molecular characterization of pepper-PVY isolated from Tehran pepper fields and it’s comparison with other PVY isolates

Detection and Characterization of Viral Species/Subspecies Using Isothermal Recombinase Polymerase Amplification (RPA) Assays

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