scholarly journals Diseño de cebadores específicos para la detección por RT-PCR del Potato Virus Y (PVY)

2019 ◽  
Vol 24 (3) ◽  
pp. 561-565 ◽  
Author(s):  
Andrea Sierra Mejía ◽  
Yuliana Marcela Gallo García ◽  
Pablo Andrés Gutiérrez Sánchez ◽  
Mauricio Alejandro Marín Montoya
Keyword(s):  
Solanum Tuberosum ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Rt Pcr

El Potato virus Y (PVY) es uno de los virus más limitantes para la producción de papa (Solanum tuberosum y S. phureja) en el mundo. Este virus es transmitido por tubérculo-semilla de papa y por diferentes especies de áfidos. Para su manejo es fundamental la siembra de tubérculos certificados por su sanidad viral, para lo que se requieren metodologías de detección altamente sensibles como ELISA y RT-PCR. Para éstas últimas pruebas, es necesario disponer de cebadores específicos que permitan el diagnóstico del virus en tejidos asintomáticos. En este estudio se reportan los cebadores PVY_Col para la detección del PVY en RT-PCR convencional y en tiempo real (RT-qPCR). Estos cebadores fueron diseñados con base en las secuencias de este virus que se han reportado en Colombia sobre diferentes hospedantes, así como de las diferentes variantes encontradas en el mundo. Una particularidad adicional de estos cebadores es que no presentan reacción cruzada con el genoma del Potato virus V (PVV), otro potyvirus que recientemente se ha encontrado afectando cultivos de papa en Colombia. Se espera que los cebadores PVY_Col sean utilizados para apoyar los programas de certificación de material de siembra de papa, así como para adelantar estudios epidemiológicos y de manejo fitosanitario de este virus.

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

2016 ◽  
Vol 34 (3) ◽  
pp. 317-328 ◽  
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.

scholarly journals Potato Virus Y (PVY) y Potato Yellow Vein Virus (PYVV) en Infecciones Mixtas No Causan Síntomas Atípicos en Plantas de Papa

2016 ◽  
Vol 11 (2) ◽  
pp. 26 ◽  
Author(s):  
Jenny Paola Alfaro García ◽  
Liliana Franco-Lara
Keyword(s):  
Solanum Tuberosum ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Yellow Vein ◽  
Solanum Phureja ◽  
Rt Pcr ◽  
Potato Yellow Vein Virus

Observaciones anteriores en cultivos de papa (<em>Solanum tuberosum</em> y <em>Solanum phureja</em>) mostraron síntomas llamados atípicos en este trabajo, que consistían en hojas con manchas irregulares verde oscuro sobre un fondo amarillo intenso, que sugieren la presencia de virus. Estos síntomas no correlacionan con ningún virus descrito para papa en Colombia. En plantas, existen reportes de infecciones virales con dos o más virus que llevan a interacciones como sinergismo o antagonismo. En este trabajo se evaluó la hipótesis de que los síntomas atípicos podrían ser resultado de infecciones mixtas entre PYVV (<em>Potato yellow vein virus) (Crinivirus) </em>y PVY (<em>Potato virus Y</em>) (<em>Potyirus)</em>, pues ambos virus son comunes en cultivos de papa en Colombia. Se reportan resultados de RT-PCR (reacción en cadena de la polimerasa mediada por retrotranscripción) para la detección de PYVV y PVY, y de ELISA para PVX, PVS, PVM y PRLV en 57 plantas con y sin síntomas atípicos provenientes de campo. Los resultados no apoyan la hipótesis planteada, pues de 10 plantas con síntomas atípicos evaluadas solo una estaba infectada con los dos virus.  Por otro lado, coinfecciones de PVY y PYVV se observaron en plantas sin síntomas aparentes (4 plantas de 5 evaluadas) y en plantas con síntomas característicos de PYVV (17 plantas de 37 evaluadas). Del total de 20 plantas evaluadas por ELISA, 18 presentaban infección por PVX aunque no se observaron síntomas asociados a este virus. Ocho de estas plantas, además de PVX estaban infectadas también con PVY y PYVV, pero mostraban síntomas de característicos de PYVV, lo que sugiere que PVX tampoco correlaciona con los síntomas atípicos. Se sugiere la presencia de un virus no reportado infectando el cultivo de papa en Colombia.

scholarly journals Diagnóstico serológico y molecular del Potato leafroll virus (PLRV) en tubérculos-semilla de papa en Antioquia, Colombia

2015 ◽  
Vol 65 (2) ◽  
pp. 204-210 ◽  
Author(s):  
Mauricio Esteban Mesa Medina ◽  
Martha Isabel González Ramírez ◽  
Pablo Andrés Gutiérrez Sánchez ◽  
Mauricio Alejandro Marín Montoya
Keyword(s):  
Solanum Tuberosum ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Potato Leafroll Virus ◽  
Rt Pcr ◽  
Potato Virus S ◽  
Leafroll Virus ◽  
Das Elisa

<p>Las virosis son uno de los problemas fitosanitarios más limitantes para la producción de papa, siendo los virus Potato leafroll virus (PLRV), Potato virus Y (PVY), Potato virus S (PVS) y Potato ye-llow vein virus (PYVV) los que generan mayores pérdidas económicas en la región andina. Gene-ralmente estos virus son transmitidos por insectos hemípteros y por tubérculos-semilla. En esta investigación se evaluó la presencia y distribución del PLRV en tubérculos de <em>Solanum tuberosum</em> variedad Diacol-Capiro y S. phureja variedad Criolla-Colombia obtenidos en Antioquia, utilizando pruebas serológicas y moleculares. Mediante DAS-ELISA se evaluaron 128 muestras consisten-tes en tres tipos de tejidos (piel, yemas latentes y brotes), mientras que con RT-PCR en tiempo real (RT-qPCR), se utilizó un subgrupo de 12 muestras de cada variedad. Utilizando DAS-ELISA, el PLRV se encontró en el 3,1% y 10,1% de las muestras de Diacol-Capiro y Criolla-Colombia, respectivamente; siendo evidente una mayor detección del virus en la piel de los tubérculos de ésta última variedad. Las pruebas de RT-qPCR detectaron el PLRV en 83,3% y 91,6% de las 12 muestras de Diacol-Capiro y de Criolla-Colombia, respectivamente; con valores de Ciclo umbral (CT) entre 17,72 y 30,59. La naturaleza de los amplicones obtenidos por RT-qPCR fue confirmada por secuenciación como parte de una región codificante de la cápside del PLRV (Identidad=99%-100%). Estos resultados enfatizan en la necesidad de utilizar tubérculo-semilla certificados por su sanidad viral en los cultivos de papa del país y de fortalecer dichos programas a partir del empleo de técnicas de detección altamente sensibles como RT-qPCR.</p>

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

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
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.

scholarly journals Arbuscular Mycorrhiza Changes the Impact of Potato Virus Y on Growth and Stress Tolerance of Solanum tuberosum L. in vitro

2020 ◽  
Vol 10 ◽  
Author(s):  
Edyta Deja-Sikora ◽  
Anita Kowalczyk ◽  
Alina Trejgell ◽  
Adriana Szmidt-Jaworska ◽  
Christel Baum ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Arbuscular Mycorrhiza ◽  
Stress Tolerance ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Solanum Tuberosum L ◽  
The Impact

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

Plant Disease ◽  
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.

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

Plant Disease ◽  
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.

Potato virus Y (PVY) detection in a single aphid by one-step RT-PCR with boiling technique

2016 ◽  
Vol 46 (4) ◽  
pp. 278-285 ◽  
Author(s):  
Juil Kim ◽  
Deok Jea Cha ◽  
Min Kwon ◽  
Rameswor Maharjan
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Rt Pcr ◽  
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