First Identification of an Unusual Recombinant Potato virus Y Strain in Potato in Mexico

Potato virus Y (PVY) has been reported in potato crops in Mexico (3), with tobacco necrotic variants found in the central State of Mexico (4). Nevertheless, many individual states are currently declared PVY free and distribution of individual strains of PVY in potato in different states of Mexico and in different solanaceous crops had not yet been studied. A limited field PVY survey was conducted on potato in the State of Chihuahua in August 2009. More than 900 random potato leaf samples were collected from cvs. Snowden, Atlantic, FL1867, Felsina, Fianna, Gigant, and Alpha. Seven were found to be PVY-positive and had been collected from cvs. Fianna, Snowden, and FL1867. The PVY status of the collected samples was initially determined with the PVY-specific Immunostrips (Bioreba, Reinach, Switzerland) and by double-antibody sandwich-ELISA using the polyclonal PVY detection kit (Agdia, Elkhart, IN). To determine the strain specificity of these PVY isolates following ELISA tests, the infected original samples were inoculated onto tobacco plants at the four-leaf stage and symptom appearance and development were observed for 8 weeks side-by-side with control isolates PB-Oz (PVYO), N4 (PVYNTN), and Mont (PVYN) (1), followed by the standard PVY strain typing by reverse transcription (RT)-PCR (2). Only one of the PVY-positive samples, originally from symptomless potato cv. Fianna, induced systemic PVY infection in tobacco by producing stunting, mosaic, and vein clearing. No systemic vein necrosis, characteristic of isolates Mont and N4, was observed in Nicotiana tabacum cvs. Burley, Xanthi, or Samsun after inoculation with this isolate during all 8 weeks of observation. This isolate, PVY-M3, was typed as a PVY recombinant by RT-PCR, with two recombinant junctions characteristic of European PVYNTN strains (2). It was further analyzed by triple-antibody sandwich-ELISA using four PVYO and PVYN strain-specific monoclonal antibodies. Monoclonals 1F5 (Agdia) and SASA-N (Scottish Agriculture Science Agency [SASA], Edinburgh) reacted to this isolate and identified PVY-M3 serologically as PVYN serotype, characteristic of other PVYNTN recombinants. Monoclonals MAb2 (Agdia) and SASA-O (SASA), specific to PVYO and PVYC strains, did not react to PVY-M3. Taken together, the combination of biological, serological, and molecular characteristics define this recombinant isolate from Mexico as belonging to the same PVY strain group represented by the isolate PVY-L26 (1). To our knowledge, this is the first report of such an unusual PVYNTN recombinant strain from Mexico. Presence of this isolate, with no vein necrotic symptoms induced on tobacco and with PVYNTN genome, will necessitate development of new detection methods for the seed potato industry in Mexico. References: (1) X. Hu et al. Virus Res. 143:68, 2009. (2) J. L. Lorenzen et al. Plant Dis. 90:935, 2006. (3) L. P. Moreno et al. Rev. Mex. Fitopatol. 22:187, 2004. (4) V. R. Ramirez-Rodriguez et al. Virol. J. 6:48, 2009.

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Prevalence and strain composition of potato virus Y circulating in potato fields in China’s north-central province Shanxi

Plant Disease ◽

10.1094/pdis-09-21-1950-re ◽

2021 ◽

Author(s):

Pengcheng Ding ◽

Dexin Chen ◽

Haixu Feng ◽

Jiao Li ◽

Hui Cao ◽

...

Keyword(s):

Potato Virus ◽

Potato Virus Y ◽

Potato Production ◽

Central China ◽

Shanxi Province ◽

Rt Pcr ◽

Potato Leaf ◽

Single Strain ◽

North Central ◽

Production Areas

Potato is an important crop in Shanxi province located in north-central China. During 2019-2020, 319 potato leaf samples were collected from eight locations distributed in three major potato production areas in Shanxi. Bio-chip detection kit revealed the presence of several potato viruses, and among them potato virus Y (PVY) was the most common one, reaching the incidence of 87.8% of all symptomatic samples. The immuno-captured multiplex reverse transcription (RT)-PCR was used to identify strains for all 280 PVY-positive samples, unveiling 242 samples infected with a single strain of PVY (86.4%) and 38 (13.6%) with a mixed infection. Of samples with a single-strain infection, PVY -SYR-II accounted for 102 (42.1%), followed by PVYN-Wi (33, 13.6%) , PVY -SYR-I (28, 11.6%), 261-4 (22, 9.1%), PVYNTNa (20, 8.3%), PVYNTNb (19, 7.9%), and PVY -SYR-III (18, 7.4%). Seven isolates representing different recombinants were selected for whole genome sequencing. Phylogenetic and recombination analyses confirmed the RT-PCR based strain typing for all seven strains of PVY found in Shanxi. SXKL-12 is the first SYR-III strain from potato reported from China. However, unlike that in other known SYR-III isolates, the region positioned from 1,764 to1,902 nt in SXKL-12 shared the highest sequence identity of 82.2% with an uncharacterized PVY isolate, JL-23, from China. Interestingly, the PVYN-Wi isolate SXZY-40 also possessed a more divergent sequence for the region positioned from 6,156 to 6,276 nt than other N-Wi isolates known to date, sharing the highest identity of 86.6% with an uncharacterized Chinese PVY isolate, JL-11. Pathogenicity analysis of dominant strains PVY -SYR-II and PVYN-Wi in six local popular potato cultivars revealed that Kexin 13, Helan 15 and Jizhangshu 12 were susceptible to these two strains with mild mottling or mosaic symptoms expression, while three cultivars, Jinshu 16, Qingshu 9, Xisen 6 were found fully resistant.

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An Improved Multiplex IC-RT-PCR Assay Distinguishes Nine Strains of Potato virus Y

Plant Disease ◽

10.1094/pdis-02-13-0161-sr ◽

2013 ◽

Vol 97 (10) ◽

pp. 1370-1374 ◽

Cited By ~ 31

Author(s):

Mohamad Chikh-Ali ◽

Stewart M. Gray ◽

Alexander V. Karasev

Keyword(s):

Potato Virus ◽

Potato Virus Y ◽

Rna Extraction ◽

Leaf Tissue ◽

Rt Pcr ◽

Pcr Assay ◽

Potato Leaf ◽

Extraction Step ◽

First Time ◽

Virus Testing

A multiplex reverse-transcription polymerase chain reaction (RT-PCR) assay was previously developed to identify a group of Potato virus Y (PVY) isolates with unusual recombinant structures (e.g., PVYNTN-NW and SYR-III) and to differentiate them from other PVY strains. In the present study, the efficiency of this multiplex RT-PCR assay was validated and extended considerably to include five additional strains and strain groups not tested before. To make the multiplex RT-PCR assay more applicable and suitable for routine virus testing and typing, it was modified by replacing the conventional RNA extraction step with the immunocapture (IC) procedure. The results obtained using well-characterized reference isolates revealed, for the first time, that this multiplex RT-PCR assay is an accurate and robust method to identify and differentiate the nine PVY strains reported to date, including PVYO (both PVYO and PVYO-O5), PVYN, PVYNA-N, PVYNTN, PVYZ, PVYE, PVY-NE11, PVYN-Wi, and PVYN:O, which is not possible by any of the previously reported RT-PCR procedures. This would make the IC-RT-PCR procedure presented here a method of choice to identify PVY strains and assess the strain composition of PVY in a given area. The IC-RT-PCR protocol was successfully applied to typing PVY isolates in potato leaf tissue collected in the field.

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Reverse Transcription Loop-Mediated Isothermal Amplification of DNA for Detection of Potato virus Y

Plant Disease ◽

10.1094/pd-89-0605 ◽

2005 ◽

Vol 89 (6) ◽

pp. 605-610 ◽

Cited By ~ 92

Author(s):

Xianzhou Nie

Keyword(s):

Potato Virus ◽

Reverse Transcription ◽

Potato Virus Y ◽

Dna Amplification ◽

Isothermal Amplification ◽

Rt Pcr ◽

Potato Leaf ◽

Time Saving ◽

Loop Mediated Isothermal Amplification ◽

One Step

A reverse transcription loop-mediated isothermal amplification of DNA (RT-LAMP) for detection of Potato virus Y (PVY) was developed. In this procedure, a set of four primers matching a total of six sequences of the coat protein (CP) gene of PVY were designed in such a way that a loop could be formed and elongated during DNA amplification. Using PVY CP complementary DNA clones as templates, the LAMP reaction was optimized by adjusting the concentrations of MgSO4, dNTPs, and Bst DNA polymerase. The effects of fragment length of target DNA on LAMP also were investigated. Two-step and one-step RT-LAMPs were performed using RNA extracts of various PVY cultures, and the results were correlated with two-step reverse transcription polymerase chain reaction (RT-PCR) for detection of PVY. Further, the turbidity caused by precipitation of magnesium pyrophosphate formed in positive RT-LAMP reactions was used to measure the amplification by utilizing a time-saving spectrophotometric method. The one-step RT-LAMP-turbidity method gave results comparable with the two-step RT-PCR method for detection of PVY from potato leaf and tuber samples. Of the total 240 samples, 234 were diagnosed similarly by both methods.

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Ocorrência de vírus em batata em sete estados do Brasil

Horticultura Brasileira ◽

10.1590/s0102-05362009000400015 ◽

2009 ◽

Vol 27 (4) ◽

pp. 490-497 ◽

Cited By ~ 7

Author(s):

Antônio Carlos de Ávila ◽

Paulo Eduardo de Melo ◽

Lindolfo R Leite ◽

Alice K Inoue-Nagata

Keyword(s):

Potato Virus ◽

Potato Virus Y ◽

Leaf Roll ◽

Potato Virus X ◽

Potato Leaf Roll Virus ◽

Rt Pcr ◽

Potato Leaf ◽

Potato Virus S ◽

Das Elisa

As viroses causam rápida degenerescência dos tubérculos-sementes de batata. Em condições tropicais, em que a presença de afídeos vetores é constante e a estrutura das populações de vírus é dinâmica, a pressão das doenças é enorme. Conhecer essa dinâmica é uma ferramenta importante para a sustentabilidade da produção de batata. Realizou-se um levantamento abrangente da ocorrência de viroses em batata no Brasil, além de estudar-se a distribuição das estirpes de Potato virus Y (PVY) associadas ao mosaico da batata. Em 2005 e 2006 foram visitadas lavouras em sete estados brasileiros, coletando-se folíolos com sintomas de viroses (1.256 amostras) e amostras aleatórias (360 amostras). Foi feita também uma estimativa visual da incidência de mosaico e enrolamento-das-folhas em vários dos campos visitados. Das 1.256 amostras suspeitas, 840 apresentaram reação positiva em teste sorológico para PVY (66,9%), 128 para Potato leaf roll virus (PLRV) (10,2%), 79 para Potato virus S (PVS) (6,3%) e nenhuma para Potato virus X (PVX). Os resultados dos testes de detecção por DAS-ELISA, biológico e RT-PCR mostraram a presença quase absoluta do subgrupo necrótico de PVY, em sua maioria PVY NTN. A análise de uma sub-amostragem em todos os municípios visitados confirmou que essa variante está hoje presente nos sete estados visitados. Amostras de PVY NTN foram obtidas das cultivares Asterix, Atlantic, Agata, Achat, Baronesa, Baraka, Bintje, Caesar, Cupido, Marijke, Monalisa, Panda e Vivaldi, que apresentaram diferentes níveis de suscetibilidade. As amostras aleatórias revelaram um quadro muito similar ao encontrado com as amostras sintomáticas. PLRV foi identificado em MG, BA, PR e SC, em várias lavouras de forma muito freqüente. PVS foi identificado nesses mesmos estados e também em SP. PVX foi detectado em apenas uma amostra tomada ao acaso em Serra do Salitre (MG). O contraste entre a avaliação visual dos sintomas e os resultados do teste de detecção por ELISA revelou a possibilidade de infecção latente por PVY em níveis relevantes na cultivar Asterix.

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Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽

10.1094/pdis-03-12-0283-re ◽

2013 ◽

Vol 97 (5) ◽

pp. 641-644 ◽

Cited By ~ 17

Author(s):

Manphool S. Fageria ◽

Mathuresh Singh ◽

Upeksha Nanayakkara ◽

Yvan Pelletier ◽

Xianzhou Nie ◽

...

Keyword(s):

Real Time ◽

Potato Virus ◽

Potato Virus Y ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Pcr Assay ◽

Current Season ◽

Seed Market ◽

Polymerase Chain ◽

Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

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Detection and genome characterization of Potato virus Y isolates infecting potato (Solanum tuberosum L.) in La Union (Antioquia, Colombia)

Agronomía Colombiana ◽

10.15446/agron.colomb.v34n3.59014 ◽

2016 ◽

Vol 34 (3) ◽

pp. 317-328 ◽

Cited By ~ 4

Author(s):

Pablo Gutiérrez S. ◽

Mauricio Marín M. ◽

Daniel Muñoz E.

Keyword(s):

Solanum Tuberosum ◽

Potato Virus ◽

Potato Virus Y ◽

Seed Tuber ◽

Rt Pcr ◽

Potato Plants ◽

Nextgeneration Sequencing ◽

Prevalent Strain ◽

Detailed Molecular Analysis

Potato virus Y (PVY) is one of the most severe viruses affecting the production of potato (Solanum tuberosum) in the world. This study presents a detailed molecular analysis using nextgeneration sequencing (NGS), IC-RT-qPCR and RT-PCR on the PVY isolates infecting seed-tubers and foliage of potato plants cv. Diacol-Capiro in La Union (Antioquia, Colombia). Analysis of incidence by IC-RT-qPCR in 15 random leaf samples of three cultivation plots and fifteen sprouting tuber eye-buds reveal infection levels between 13.4 and 80%; a higher incidence of 86.7% was observed in seed-tuber samples with threshold cycle (Ct) values as low as 24.3. Genome assembly from a bulk of foliage samples resulted in a consensus PVY genome (PVY_LaUnionF) of 9,702 nt and 399 polymorphic sites within the polyprotein ORF; while the assembled genome from sprouts of tubers has 9,704 nt (PVY_LaUnionT) and contained only six polymorphic nucleotide sites. Phylogenetic analysis demonstrates that the PVY isolates from leaf samples are in the recombinant PVYNTN group (sequence identity >99%); while those from tuber sprouts are in the PVYN/NTN group with identities above 95%. Sanger sequencing of viral capsid suggests the presence of a third variant related to PVYO, a prevalent strain reported in potato fields worldwide.

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The Recombinant Potato virus Y (PVY) Strain, PVYNTN, Identified in Potato Fields in Victoria, Southeastern Australia

Plant Disease ◽

10.1094/pdis-05-20-0961-sc ◽

2020 ◽

Vol 104 (12) ◽

pp. 3110-3114

Author(s):

Mariana Rodriguez-Rodriguez ◽

Mohamad Chikh-Ali ◽

Steven B. Johnson ◽

Stewart M. Gray ◽

Nellie Malseed ◽

...

Keyword(s):

Potato Virus ◽

Potato Virus Y ◽

Strain Typing ◽

Rt Pcr ◽

Growing Seasons ◽

Fta Cards ◽

Southeastern Australia ◽

Whole Genomes ◽

Pcr Assays

Potato virus Y (PVY) is one of the main viruses affecting potato in Australia. However, molecular characterization of PVY isolates circulating in potato in different states of Australia has not yet been thoroughly conducted. Only nonrecombinant isolates of three biological PVY strains collected from potato were reported previously from Western Australia and one from Queensland. Here, PVY isolates collected from seed potato originating in Victoria, Australia, and printed on FTA cards, were subjected to strain typing by RT-PCR, with three isolates subjected to whole genome sequencing. All the 59 PVY isolates detected during two growing seasons were identified to be recombinants based on two RT-PCR assays. No nonrecombinant PVY isolates were identified. All the RT-PCR typed isolates belonged to the PVYNTN strain. Sequence analysis of the whole genomes of three isolates suggested a single introduction of the PVYNTN strain to Australia but provided no clues as to where this introduction originated. Given the association of the PVYNTN strain with potato tuber damage, growers in Australia should implement appropriate strategies to manage PVYNTN in potato.

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Greenhouse Comparison of Two Detection Methods for Potato virus YN-Wi at Four Potato Growth Stages

Plant Health Progress ◽

10.1094/php-11-17-0072-rs ◽

2018 ◽

Vol 19 (1) ◽

pp. 71-75 ◽

Cited By ~ 1

Author(s):

Abby Beissinger ◽

Debra Ann Inglis

Keyword(s):

Potato Virus ◽

Detection Method ◽

Sandwich Elisa ◽

Potato Production ◽

Detection Methods ◽

Growth Stages ◽

Veinal Necrosis ◽

Leaf Drop ◽

Potato Growth ◽

Common Laboratory

Potato virus Y (PVY) causes significant crop and monetary losses. Owing to the prevalence of newly emerging strains of PVY such as PVYN-Wi, which often cause asymptomatic to mild reactions on certain potato cultivars, accurate tools are required to detect the virus in potato production. This study compared the sensitivity of a rapid field detection method (immunostrips) with a common laboratory detection method (triple antibody sandwich ELISA) on cultivar Chieftain, grown under isolated conditions in a greenhouse and mechanically inoculated with PVYN-Wi, at four potato growth stages (emergence, preflower, postflower, and senescence). Plants inoculated at emergence displayed severe symptoms of mosaic, veinal necrosis, and leaf drop. Plants inoculated at preflower, postflower, and senescence had veinal necrosis but low or no incidence of mosaic and leaf drop. Overall, few or no tuber symptoms were observed, but a trend of lower tuber yield occurred for emergence-inoculated plants. Low variability in PVYN-Wi detection occurred in both tests for emergence-inoculated plants, whereas those inoculated at preflower and postflower had more variability. Because symptom expression may differ depending on the growth stage when a plant becomes infected, these variations should be heeded with either detection method when collecting samples for PVY testing.

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