scholarly journals Biological and molecular characterization of various isolates  of Potato virus Y-N (PVY-N) strain group

2017 ◽  
Vol 38 (SI 2 - 6th Conf EFPP 2002) ◽  
pp. 278-280
Author(s):  
J. Ptáček ◽  
P. Dědič ◽  
J. Matoušek

Fourteen Potato virus Y (PVY) isolates were characterized. They represented PVYN strain only. However, application of serological and molecular genetic methods led to a more complicated characterization. For example, five isolates induced necrotic symptoms on tobacco plants typical of PVY<sup>N</sup>, despite reacting as PVY<sup>O</sup> serologically. Moreover, the PVY isolates were not identical according to molecular genetic properties. Typical PVY<sup>NTN</sup> PCR products were observed for 11 isolates, but four of them (Hr220-5, Hr387-7, Nord 242 and Syn1Scot) did not produce potato tuber necrotic symptoms in infected cultivars. An immunocapture RT-PCR probing was developed using a set of 24 primer pairs derived from eight regions of the PVY genome. Using this method, five out of seven PVY<sup>NTN</sup> isolates including the Czech standard PVY<sup>NTN</sup> from the potato cv. Nicola were found to be identical. However, two PVY<sup>NTN</sup> isolates and all the other probed PVY samples showed unique patterns, suggesting specific differences at the nucleotide level. This method enabled specific identification of individual isolates variability even within different PVY strains.

Plant Disease ◽  
2008 ◽  
Vol 92 (9) ◽  
pp. 1371-1371 ◽  
Author(s):  
A. V. Karasev ◽  
T. Meacham ◽  
X. Hu ◽  
J. Whitworth ◽  
S. M. Gray ◽  
...  

Potato virus Y (PVY) causes substantial losses in potato production by decreasing yields and affecting the quality of potato tubers. Management of PVY in potato is dependent primarily on potato seed certification programs to prevent or limit initial levels of virus inoculum. Prior to 1990, the ordinary strain of PVY (PVYO) was the predominant virus in North America. PVYO induces clear foliar symptoms in many potato cultivars, allowing successful management in seed potato through a combination of visual inspections and limited laboratory testing. In recent years, necrotic strains of PVY (PVYN, PVYNTN, and PVYN:O) have begun to spread in the United States, many of which induce mild symptoms in potato, making them more difficult to manage through visual inspections. In addition to reducing yield, necrotic isolates may also cause external and internal damage in tubers of susceptible cultivars, which is known as potato tuber necrotic ringspot disease (PTNRD). Tuber necrotic strains of PVY have been reported across the northern United States (1,2,4), although limited information is available on their incidence and spread in commercial potato production. During June and July of 2007, 38 random samples were collected from three different commercial fields displaying disease problems (cvs. Russet Ranger, Alturas, and Russet Burbank) in the vicinity of Idaho Falls, ID. Plants collected showed various degrees of mosaic and leaf yellowing. By using double-antibody sandwich (DAS)-ELISA and reverse transcription (RT)-PCR, 25 of these plants were identified as PVY positive. The mutiplex RT-PCR assay (3) confirmed that nine plants were infected with PVYNTN and 11 with PVYN:O. No RT-PCR products were amplified from five samples. During September and October of 2007, 25 tuber samples (cv. Russet Burbank) showing various degrees of unusual internal symptoms (e.g., brown spots) were collected near Idaho Falls, ID. Twenty-two tubers were found PVY positive by DAS-ELISA, and multiplex RT-PCR determined 13 of those were PVYNTN, three were PVYO, one was a PVYNTN/N:O mixture, and one was a PVYO/N:O mixture. No RT-PCR products were amplified from four samples. In October 2007, six tubers showing distinct external tuber damage characteristic of PTNRD (cv. Highland Russet) were collected near Twin Falls, ID. All six tubers were determined to be PVY positive by DAS-ELISA, and RT-PCR identified five as infected with PVYNTN and one with PVYN:O. All the mixtures were easily separated by inoculating tobacco plants followed by subsequent testing of individual plants. Asymptomatic tubers from the same lot not showing PTNRD damage were found PVY negative by DAS-ELISA and RT-PCR. All PVYNTN isolates collected during 2007 were inoculated into tobacco plants (Nicotiana tabacum L. cv. Xanthi) and confirmed to induce systemic vein necrosis. Limited sequencing of four of the PVYNTN isolates determined that they contained recombinant junctions 2 and 3, identifying them as being related to the European strain of PVYNTN (3). The data suggest an increase in distribution and incidence of necrotic strains of PVY in commercial, potato-production areas in Idaho during an outbreak in 2007 and the potential for an increase in PTNRD. References: (1) P. M. Baldauf et al. Plant Dis. 90:559, 2006. (2) J. M. Crosslin et al. Plant Dis. 90:1102, 2006. (3) J. H. Lorenzen et al. Plant Dis. 90:935, 2006. (4) L. M. Piche et al. Phytopathology 94:1368, 2004.


2016 ◽  
Vol 34 (3) ◽  
pp. 317-328 ◽  
Author(s):  
Pablo Gutiérrez S. ◽  
Mauricio Marín M. ◽  
Daniel Muñoz E.

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.


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 ◽  
...  

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.


1987 ◽  
Vol 38 (2) ◽  
pp. 395 ◽  
Author(s):  
R Heath ◽  
RJ Sward ◽  
JR Moran ◽  
AJ Mason ◽  
ND Hallam

Six potato virus Y isolates from Victoria and Queensland were characterized on the basis of host plant reactions. Four isolates from potato produced symptoms in indicator plants and potato cultivars consistent with those caused by the PVYO group. Two isolates from tobacco produced necrotic symptoms on some tobacco cultivars characteristic of the PVYN group, but did not systemically infect potato cultivars and therefore could not be considered to belong to any of the previously described PVY strain groups. Despite the wide variation in their biological characteristics, an antiserum produced to one of the potato isolates was able to detect all six of the isolates when used in ELISA


Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1177-1177 ◽  
Author(s):  
J. M. Crosslin ◽  
P. B. Hamm ◽  
K. C. Eastwell ◽  
R. E. Thornton ◽  
C. R. Brown ◽  
...  

More than 50 isolates of Potato virus Y (PVY) with characteristics of strains that cause tobacco veinal necrosis (PVYN) were obtained from potatoes (Solanum tuberosum L.) grown in the northwestern United States. These isolates are being characterized at the biological and molecular levels. Isolate RR1 was obtained from leaves of potato cv. Ranger Russet showing distinct mottling and leaf deformity, which is in contrast to the leaf-drop and necrosis usually observed with ordinary strains of PVY (PVYO) in this variety. Isolate AL1 was obtained from tubers of potato cv. Alturas showing distinct internal light brown rings and blotches. When RR1 and AL1 were transmitted to tobacco (Nicotiana tabacum L. cvs. Samsun NN and 423), they caused systemic veinal necrosis, including stem and petiole lesions typical of PVYN strains (2). Symptoms induced by RR1 and AL1 on tobacco appeared 9 to 11 days after inoculation, whereas some other isolates caused delayed veinal necrosis. All isolates that produced veinal necrosis on tobacco were detectable with PVY polyclonal antisera. Potato virus X was not detected by enzyme-linked immunosorbent assay in tobacco plants showing veinal necrosis. Some isolates, including AL1, failed to react in serological tests using PVYN-specific monoclonal antibodies obtained from three commercial sources. Other isolates, including RR1, were detectable with these monoclonal antibodies. Reverse transcription-polymerase chain reaction (RT-PCR) products obtained with primers specific for the coat protein (CP) open reading frame (ORF) were cloned and sequenced. AL1 possesses a CP more closely related to PVYO type isolates, which would account for its failure to react with PVYN monoclonal antibodies. In this regard, AL1 is similar to the PVYN-Wilga isolate (1). Other isolates that are detectable with the PVYN monoclonal antibodies possess a CP more consistent with N strains of the virus. Results of RT-PCR tests using primers derived from the P1 ORF sequence (3), and the restriction enzyme analysis and sequencing of the RT-PCR products, all suggest that AL1 and RR1 are related to European-type members of PVY tuber necrotic (NTN) or N strains. However, other isolates under investigation appear to be more closely related to previously reported North American NTN types (3). The symptomatology of these viruses on tobacco and potato, and the serological and molecular data clearly show that at least two distinct variants of PVYN have been found for the first time in a major potato production area of the United States, and pose a potential threat to the potato industry. References: (1) B. Blanco-Urgoiti et al. Eur. J. Plant Pathol. 104:811, 1998. (2) J. A. de Bokx and H. Huttinga. Potato virus Y. Descriptions of Plant Viruses. No. 242, CMI/AAB, Surrey, England, 1981. (3) R. P. Singh et al. Can J. Plant Pathol. 20:227, 1998.


Plant Disease ◽  
2019 ◽  
Vol 103 (1) ◽  
pp. 137-142 ◽  
Author(s):  
Mohamad Chikh-Ali ◽  
Mariana Rodriguez-Rodriguez ◽  
Kelsie J. Green ◽  
Dong-Jun Kim ◽  
Sang-Min Chung ◽  
...  

Potato is an important source of food in South Korea, and viruses represent a significant threat to sustainable and profitable potato production. However, information about viruses affecting the potato crop in South Korea is limited. In 2017, potato plants of five cultivars exhibiting foliar mosaic, crinkling, and mottle were collected in two seed potato production areas, in Gangwon-do and Jeollabuk-do Provinces, and subjected to virus testing and characterization. Potato virus Y (PVY) was found associated with mosaic symptoms, and samples were characterized using reverse transcription polymerase chain reaction (RT-PCR) and whole genome sequencing. All analyzed PVY-positive samples were found to represent the same recombinant PVY strain: PVYNTN. Three PVY isolates were subjected to whole genome sequencing using overlapping RT-PCR fragments and Sanger methodology, and all three were confirmed to represent strain PVYNTNa after a recombination analysis of the complete genomes. In phylogenetic analysis, the three South Korean isolates were placed most closely to several PVYNTNa isolates reported from Japan and Vietnam, suggesting a common source of infection. This is the first report and complete molecular characterization of a PVYNTN strain present in the country, and because this strain induces tuber necrotic ringspot disease in susceptible cultivars of potato, appropriate management tools need to be implemented to mitigate potential tuber quality losses.


Plant Disease ◽  
2008 ◽  
Vol 92 (11) ◽  
pp. 1586-1586 ◽  
Author(s):  
H. Xu

Most strains of Potato virus Y (PVY) can infect tobacco plants (Nicotiana tabacum) and cause vein clearing followed by leaf mottling, except the PVYN strain, which induces severe vein necrosis. Some isolates within the PVYN strain also cause potato necrotic tuber ringspot disease, but these have not been reported from Canadian tobacco fields. PVYNTN isolates include European (EU) and North American (NA) types that are serologically identical to PVYN, but can be distinguished by nucleic acid-based assays and potato bioassay (1,2). Some PVY isolates, PVYN-Wi or PVYN:O, resulting from a recombination between RNA molecules of PVYN and the common strain, PVYO, are identified as PVYO in serological assays, but induce necrosis in tobacco (2). In August of 2007, two samples of tobacco (N. tabacum, unknown cultivar) leaves showing necrotic symptoms resembling those induced by PVYN, PVYNTN, or PVYN-Wi were collected from a tobacco field in southern Ontario, Canada and submitted to the Charlottetown Laboratory, Canadian Food Inspection Agency, Charlottetown, PE. Virus in both samples (PVY-204 and PVY-205) reacted with PVYN-specific antibodies 1F5 and 4E7 (3) and induced vein necrosis in tobacco (N. tabacum cv. Samsun). A multiplex reverse transcription (RT)-PCR assay (1) for the simultaneous detection and differentiation of various PVY strains amplified two fragments (181 and 452 bp) associated with EU-PVYNTN isolates. Restriction fragment length polymorphism (RFLP) analysis targeting the P1 and NIb gene (3) also indicated that PVY-204 and PVY-205 were EU-PVYNTN isolates. Known isolates of PVYO, PVYN, and NA-PVYNTN were used in all evaluations as references (3). Furthermore, the nucleotide sequences of the P1 and NIb genes of PVY-204 and PVY-205 determined by automated cycle sequencing (3) and subjected to phylogenetic analysis indicated that the nucleotide and deduced amino acid sequences of both isolates were 96 and 95% identical, respectively, to NA-PVYNTN isolates reported from Canada, but 99% identical (both nucleotide and amino acid) to EU-PVYNTN isolates from Europe and Mexico (3). Potato (cv. Yukon Gold) plants mechanically inoculated with leaf sap from tobacco (N. tabacum cv. Samsun) infected with PVY-204 and PVY-205 developed various leaf symptoms including severe local and systemic necrotic lesions, leaf wilting, and leaf death in 3 to 5 weeks postinoculation under greenhouse conditions. The infected plants recovered in 5 to 6 weeks. Potato (cv. Yukon Gold) plants inoculated with leaf sap from tobacco (N. tabacum cv. Samsun) infected with a PVYNTN isolate (HX8) (3) and healthy tobacco leaf sap were used as positive and negative controls. The number and yield of the tubers harvested from infected plants were significantly reduced (50%), and PVY-204 and PVY-205 induced typical potato tuber necrotic ringspot disease in 52.6% of the progeny tubers with an average disease index of 0.364 (C. Kerlan and K. Charlet-Ramage, EAPR Virology Meeting Proceedings, 1998). PVYNTN was detected by RT-PCR and RFLP in all necrotic tubers and 66.7% of the asymptomatic tubers. Some tubers (15.8%) harvested from the infected plants were negative in RT-PCR targeting either P1 protein gene or NIb gene and showed neither external nor internal necrotic symptom. To my knowledge, this is the first evidence of the occurrence of PVYNTN isolates in field-grown tobacco plants in Canada. References: (1) J. H. Lorenzen et al. Plant Dis. 90:935, 2006. (2) R. Singh et al. Arch Virol. 153:1, 2008. (3) H. Xu et al. Can. J. Plant Pathol. 27:125, 2005.


Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  

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.


Sign in / Sign up

Export Citation Format

Share Document