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 ◽  
...  

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.

Plant Disease ◽  
2013 ◽  
Vol 97 (3) ◽  
pp. 430-430 ◽  
Author(s):  
A. Quintero-Ferrer ◽  
A. V. Karasev

Potato virus Y (PVY) is a serious problem for potato production worldwide. The virus reduces both tuber yield and quality, and recent spread of recombinant strains of PVY in potato production areas is largely credited with the spread of potato tuber necrotic ringspot disease (PTNRD) (1). In Mexico, recombinant strains of PVY were reported in at least two states, Chihuahua (4) and the State of Mexico (3); however, no surveys have been conducted in other potato-producing areas, and the spectrum of PVY isolates circulating in the country has remained uncharacterized. In October 2011, a small-scale survey of seed potato was conducted in the state of Jalisco, Mexico, to identify PVY isolates present in fields. Twelve seed potato fields were inspected visually. These represented various generations of seed potato, from nuclear to G2. Leaf samples were collected from plants displaying mosaic, crinkling, and yellowing symptoms, and were tested for PVY. Fifty samples were collected from cultivars Fabula, Mondial, Fianna, Gigant, Caesar, and Adora. Of the 50 leaf samples collected, seven were PVY-positive using the Immuno-strip Kit (Agdia, Elkhart, IN), and six of these were determined to have a N-serotype according to the typing by the Pocket Diagnostics lateral flow kit (Forsite Diagnostics, Ltd., York, UK). PVY-positive samples came from cultivars Fabula (2 with N serotype), Mondial (4 with N serotype), and Fianna (1 with O serotype). Extracts of the seven PVY-positive leaf samples were applied to Whatman FTA cards (Sigma, St. Louis, MO), dried, and transported to the Plant Virology Laboratory at the University of Idaho for further characterization. All samples immobilized on FTA cards were subjected to RNA extraction and standard reverse transcriptase (RT)-PCR typing using a set of PVY-specific primers (2) to determine the strain type. All PVY isolates were recombinant. The six N-serotype samples were found to contain recombinant PVYNTN isolates and produced characteristic bands of 181 and 452 bp in RT-PCR, which indicated the presence of two recombination junctions in the HC-Pro/P3 and VPg regions typical of European PVYNTN isolates. The one O-serotype sample was identified as a recombinant PVYN-Wi/N:O isolate, and produced 181 and 689 bp bands in RT-PCR, which indicated the presence of one recombination junction in the HC-Pro/P3 region. Sequence analysis of RT-PCR products amplified from five samples with N serotype identified them as PVYNTN isolates, and from the one with O serotype identified it as PVYN-Wi/N:O isolate. Sequence comparisons confirmed that N serotype samples contained PVY isolates most closely related to typical PVYNTN sequences (Accession No. EF026075), while the O serotype sample contained the PVY isolate most closely related to PVYN-Wi from Europe (HE608963). The data obtained suggest the presence of two different types of PVY recombinants, PVYNTN and PVYN-Wi, in seed potato in Jalisco. Additional surveillance for these recombinant isolates may be needed, as well as a survey of their effects on tuber quality in production areas. This is the first report of recombinant isolates of PVY often associated with PTNRD circulating in seed potato in Jalisco, Mexico. References: (1) S. M. Gray et al. Plant Dis. 94:1384, 2010. (2) J. H. Lorenzen et al. Plant Dis. 90:935, 2006. (3) V. R. Ramirez-Rodriguez et al. Virol. J. 6:48, 2009. (4) L. Robles-Hernandez et al. Plant Dis. 94:1262, 2010.


Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1370-1374 ◽  
Author(s):  
Mohamad Chikh-Ali ◽  
Stewart M. Gray ◽  
Alexander V. Karasev

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.


Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 605-610 ◽  
Author(s):  
Xianzhou Nie

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.


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 ◽  
2014 ◽  
Vol 98 (2) ◽  
pp. 287-287 ◽  
Author(s):  
T. A. Damayanti ◽  
O. J. Alabi ◽  
S. H. Hidayat ◽  
J. M. Crosslin ◽  
R. A. Naidu

Potato (Solanum tuberosum) is an important vegetable crop in Indonesia. A small survey was conducted for virus diseases in November 2011 in Lembang, West Java, as part of assessing the sanitary status of potatoes produced in farmers' fields. Among the six potato fields surveyed, one field had nearly 20% of plants displaying stunted growth with leaves showing mild chlorotic spots and reduced size of lamina. Tubers harvested from symptomatic plants showed no necrosis symptoms. Symptomatic leaves from three representative potato plants were positive for Potato virus Y (PVY) when tested with PVY-specific immunostrips (Agdia Inc., Elkhart, IN). Leaf samples from virus-positive plants were imprinted on FTA Classic Cards (Whatman International Ltd., Maidstone, UK), air dried, and shipped to Washington State University for confirmatory diagnostic tests. Total nucleic acids were eluted from FTA cards (1) and subjected to reverse transcription (RT)-PCR using primers (PVY/Y4A and PVY/Y3S) specific to the coat protein (CP) of PVY (3). Nucleic acid extracts from samples infected with PVY ordinary strain (PVYO), tuber necrosis strain (PVYNTN), tobacco veinal necrosis strains (PVYEU-N and PVYNA-N), and a recombinant strain (PVYN:O) were included as standards to validate RT-PCR assays. The approximately 480-bp DNA fragment, representing a portion of the CP, amplified in RT-PCR was cloned into pCR2.1 (Invitrogen Corp., Carlsbad, CA). DNA isolated from four independent recombinant clones was sequenced from both orientations. Pairwise comparison of these sequences (GenBank Accession Nos. KF261310 to 13) showed 100% identity among themselves and 93 to 100% identity with corresponding sequences of reference strains of PVY available in GenBank (JQ743609 to 21). To our knowledge, this study represents the first confirmed report of PVY in potato in West Java, Indonesia. Studies are in progress to assess the prevalence of PVY in other potato-growing regions of Indonesia and document the presence of different strains of the virus (2). Since the majority of farmers in Indonesia plant seed selected from their previous potato crop, there is an increased risk of primary and secondary spread of PVY through the informal seed supply system, leading to its increased significance to potato production in Indonesia. Therefore, strengthening foundation seed potato and supply chain programs will promote the production of virus-free potatoes in Indonesia. References: (1) O. J. Alabi et al. Plant Dis. 96:107, 2012. (2) A. Karasev and S. M. Gray. Am. J. Potato Res. 90:7, 2013. (3) R. P. Singh et al. J. Virol. Methods 59:189, 1996.


Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 935-940 ◽  
Author(s):  
James H. Lorenzen ◽  
Lisa M. Piche ◽  
Neil C. Gudmestad ◽  
Teresa Meacham ◽  
Pat Shiel

Potato virus Y (PVY) has become a serious problem for the seed potato industry, with increased incidence and rejection of seed lots submitted for certification. New PVY strains and strain variants have emerged in recent decades in Europe and North America, including the PVYN strain that causes veinal necrosis in tobacco, and strain variants that represent one or three recombination events between the common strain (PVYO) and PVYN. Several reverse transcription-polymerase chain reaction (RT-PCR) assays have been described that characterize PVY isolates as to strain type, but they are limited in their ability to detect some combinations of mixed strain infections. We report here the development of a single multiplex RT-PCR assay that can assign PVY strain type and detect mixed infections with respect to the major strain types. Validation of this assay was achieved using 119 archived PVY isolates, which had been previously characterized by serology and bioassay and/or previously published RT-PCR assays. Results for single-strain isolates were comparable to previous results in most cases. Interestingly, 16 mixed infections were distinguished that had previously gone undetected. The new multiplex RT-PCR assay will be useful for researchers and seed production specialists interested in determining PVY infection type using a single assay.


Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1262-1262 ◽  
Author(s):  
L. Robles-Hernández ◽  
A. C. González-Franco ◽  
J. Hernández-Huerta ◽  
T. M. Meacham ◽  
O. V. Nikolaeva ◽  
...  

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.


Plant Disease ◽  
2019 ◽  
Vol 103 (2) ◽  
pp. 289-297 ◽  
Author(s):  
Yanju Bai ◽  
Shuxin Han ◽  
Yanling Gao ◽  
Wei Zhang ◽  
Guoquan Fan ◽  
...  

In 2011–2014, ELISA or nucleic acid spot hybridization (NASH) testing for common potato viruses or Potato spindle tuber viroid (PSTVd) was performed on 500 leaf samples collected in potato fields in the northeast provinces Heilongjiang and Inner Mongolia, China. The results revealed that 38.4% (Heilongjiang) and 27.7% (Inner Mongolia) were positive for Potato virus Y (PVY). To unveil the strain composition and population structure of PVY in the region, the multiplex RT-PCR described by Chikh-Ali et al. was performed on all of the ELISA-PVY-positive samples. Of the 158 samples whose PVY strain scenarios could be determined, PVYNTN-NW-SYR-II and PVYN-Wi were the most abundant strains, occurring in 58.9 and 47.5% samples, followed by PVYNTN-NW-SYR-I (31.0%), PVYN:O (19.6%), Eu-PVYNTN (7.6%), NA-PVYN (1.3%), and PVYO (0.6%). In the 84 single-strain-infected samples, PVYN-Wi accounted for 41.7%, PVYNTN-NW-SYR-II for 40.5%, PVYNTN-NW-SYR-I for 14.3%, and PVYN:O and Eu-PVYNTN for 3.6% each. Seven isolates representing PVYNTN-NW-SYR-I (HLJ-6-1 and HLJ-9-4), PVYNTN-NW-SYR-II (INM-W-369-12 and SC-1-1-2), PVYN:O (HLJ-30-2), and PVYN-Wi (HLJ-BDH-2 and HLJ-C-429) were sequenced and analyzed molecularly. Whereas the sequence identities for isolates belonging to the same strain group were >98.5%, they fell for isolates belonging to different strain groups to 92.7–98.1% at the genome level and 96.1–98.4% at the polyprotein level. Interestingly, the exact location of the recombination events varied among isolates within a strain group. Phylogenetic analysis of all 42 full length PVY sequences from China indicated that most clustered to various recombinant groups, despite the fact that the PVY isolates were isolated from at least five host species. Pathological analysis of four isolates representing PVYN:O, PVYN-Wi, PVYNTN-NW-SYR-I, and PVYNTN-NW-SYR-II revealed that the PVYNTN-NW-SYR-II isolate incited the most severe symptoms on potato cultivar Kexin 13, followed by PVYNTN-NW-SYR-I, PVYN:O and PVYN-Wi. The PVYNTN-NW-SYR-I and PVYNTN-NW-SYR-II isolates also caused necrotic ringspots on the tubers of Kexin 13, indicating their ability to induce the potato tuber necrotic ringspot disease in potato.


Plant Disease ◽  
2003 ◽  
Vol 87 (6) ◽  
pp. 679-685 ◽  
Author(s):  
R. P. Singh ◽  
D. L. McLaren ◽  
X. Nie ◽  
M. Singh

Surveys of commercial and seed potato fields for virus diseases (1998 to 2002) in Manitoba established that Potato virus Y (PVY) is of concern in seed potato production. To determine the prevalence of PVY strains, PVY-infected tubers identified by reverse transcription-polymerase chain reaction (RT-PCR) from surveys (2000 to 2001) were grown for symptom expression and strain characterization by strain-specific RT-PCR, bioassays, and serological assays. Of the samples collected (2000 to 2001) and tested by RT-PCR, 4.0% contained PVY. Further analysis of the PVY-positive samples by a duplex RT-PCR facilitating the simultaneous detection of common (PVYO) and tobacco veinal necrosis strains (PVYN/NTN) indicated that 37.5% contained PVYO and 63.5% contained PVYN-type isolates. Analysis of the PVYN-type samples using three monoclonal antibodies (MAbs) showed that all reacted with only the PVYO MAbs and not with the PVYN-specific MAb. Partial nucleotide sequences of both ends of PVY-RNA showed that the PVYN-type isolates resembled those reported in 1996 from Manitoba. These isolates are designated as PVYN:O. In view of the increased incidence of PVYN:O in one production area, seed tubers imported from other provinces of Canada and the neighboring United States were analyzed for PVYN:O. The PVYN:O was detected in imported seeds from Minnesota, Montana, and North Dakota.


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.


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