scholarly journals ELISA reagents for potato virus Y strains with significantly low non-specific reactions

1984 ◽  
Vol 56 (1) ◽  
pp. 89-95
Author(s):  
Aarne Kurppa ◽  
Kirsi Korhonen
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Secondary Infection ◽  
Protein A ◽  
Elisa Test ◽  
Leaf Sample ◽  
Potato Leaf ◽  
Specific Antisera ◽  
Specific Reactions

High titered and highly virus-specific antisera to selected PVYo and PVYn antigens and their mixture were produced in rabbits. Immunoglobulins purified from the antisera with the protein A method and their enzyme conjugates had very strong virus-specific but no non-specific reactions in the ELISA test. Their homologous reactions to the antigens were stronger than heterologous but any PVY isolate could be identified in a potato leaf sample with the dilutions between 10-2 and 10-3 and in a sample from sprouted tubers with secondary infection diluted between 10-1 and 10-2.

scholarly journals Symptom expression and accumulation of potato virus Y (PVYO) and potato leaf roll virus in thirteen potato cultivars

1993 ◽  
Vol 2 (1) ◽  
pp. 33-40
Author(s):  
Jari Valkonen ◽  
Eerik Mäkäräinen
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Secondary Infection ◽  
Leaf Roll ◽  
Symptom Expression ◽  
Potato Leaf ◽  
Systemic Necrosis ◽  
Local Lesions ◽  
Ordinary Strain ◽  
Leafroll Virus

Necrotic local lesions developed in cvs. Matilda, Ostara, Record, Satuma, Stina, Hankkija’s (Hjan) Tanu and Hjan Timo and local ring spots in Olympia and Sieglinde (Siikli) following sap inoculation with the ordinary strain of potato virus Y (PVY0). Secondarily infected cvs. Ostara, Pito, Siikli and Hjan Timo developed leaf drop. No infected progeny was produced by Matilda, Saturna and Hjan Tanu. In contrast, Bintje, Puikula and Sabina developed neither local lesions nor systemic necrosis, but showed mosaic symptoms following primary and secondary infection by PVYO. The ELISA absorbance values for potato leafroll virus (PLRV) in Ostara, Pito and Saturna were less than 10% of those in the PLRV-infected Siikli. The ELISA values for PLRV in Olympia, Stina, Hjan Tanu and Hjan Timo were not significantly different from those of Siikli. The severity of the symptoms did not correlate with the concentration of PLRV in the potatoes.

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

Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1370-1374 ◽  
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.

scholarly journals Strain-Specific Hypersensitive and Extreme Resistance Phenotypes Elicited by Potato virus Y Among 39 Potato Cultivars Released in Three World Regions Over a 117-Year Period

Plant Disease ◽  
2018 ◽  
Vol 102 (1) ◽  
pp. 185-196 ◽  
Author(s):  
Roger A. C. Jones ◽  
Stuart J. Vincent
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Secondary Infection ◽  
Potato Cultivars ◽  
Symptom Expression ◽  
Extreme Resistance ◽  
Potato Plants ◽  
Pvy Resistance ◽  
World Regions ◽  
Healthy Seed

Strain-specific hypersensitive (HR) and extreme resistance (ER) phenotypes elicited in potato plants by three Potato virus Y (PVY) isolates in strain groups PVYO (BL and DEL3) and PVYD (KIP1) were studied. PVYO and PVYD isolates elicit HR genes Ny or putative Nd, respectively, and all three isolates elicit ER gene Ry. They were inoculated to 39 Australasian, European, or North American potato cultivars released over a 117-year period and harvested tubers were replanted. Both primary and secondary symptoms were recorded. Two European cultivars always developed ER following sap and graft inoculation and, thus, carried comprehensive PVY resistance gene Ry. One Australasian and two European cultivars always developed susceptible phenotypes and, thus, lacked genes Ry, Ny, and putative Nd. Sap inoculation with isolate KIP1 elicited localized HR (LHR) in 31 cultivars and both LHR and systemic HR (SHR) in three others; thus, all carried putative Nd. Isolates BL and DEL3 both elicited susceptible phenotypes in 11 of these 34 cultivars but LHR alone, SHR alone, or both LHR and SHR in the other 23 which, therefore, all carry Ny. With these two isolates, SHR expression ranged from very severe to very weak, with the greatest numbers of isolate–cultivar combinations occurring in the severe category with BL (n = 11) and moderate category (n = 12) with DEL3. Within the same isolate–cultivar combination, overall, SHR symptom expression was weaker with secondary than primary infection. With both primary and secondary infection, SHR expression was most severe with KIP1 and weakest with DEL3. Genes Ny and putative Nd were present in cultivars released between 1939 and 2010 or 1893 and 2010, respectively, occurring in cultivars from all three world regions. These findings have important implications concerning breeding new PVY-resistant potato cultivars, especially for countries lacking healthy seed potato stocks, or where subsistence farmers cannot afford them. An alternative to including gene Ry is incorporating as many strain-specific PVY resistance genes as possible.

scholarly journals Eliminating Potato Virus Y (PVY) and Potato Leaf Roll Virus (PLRV) Using Cryotherapy of in vitro-grown Potato Shoot Tips

2014 ◽  
Vol 59 (4) ◽  
pp. 498-504 ◽  
Author(s):  
Jung-Yoon Yi ◽  
◽  
Gi-An Lee ◽  
Jong-Wook Jeong ◽  
Sok-Young Lee ◽  
...  
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Leaf Roll ◽  
Shoot Tips ◽  
Potato Leaf Roll Virus ◽  
Potato Leaf

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

Plant Disease ◽  
2005 ◽  
Vol 89 (6) ◽  
pp. 605-610 ◽  
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.

scholarly journals Potato virus Y: Contact Transmission, Stability, Inactivation, and Infection Sources

Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 387-394 ◽  
Author(s):  
B. A. Coutts ◽  
R. A. C. Jones
Keyword(s):  
Potato Virus ◽  
Potato Virus Y ◽  
Management Strategies ◽  
Previous History ◽  
Milk Powder ◽  
Weed Species ◽  
Potato Leaf ◽  
Tobacco Plants ◽  
Potato Plants ◽  
Volunteer Potatoes

In glasshouse experiments, two isolates of Potato virus Y ‘O’ strain (PVYO) were transmitted from infected to healthy potato plants by direct contact when leaves were rubbed against each other, when cut surfaces of infected tubers were rubbed onto leaves, and to a limited extent, when blades contaminated with infective sap were used to cut healthy potato tubers. However, no tuber-to-tuber transmission occurred when blades were used to cut healthy tubers after cutting infected tubers. When leaf sap from potato plants infected with two PVYO isolates was kept at room temperature, it was highly infective for 6 to 7 h and remained infectious for up to 28 h. Also, when sap from infected leaves with one isolate was applied to five surfaces (cotton, hessian, metal, rubber vehicle tire, and wood) and left to dry for up to 24 h before each surface was rubbed onto healthy tobacco plants, PVYO remained infective for 24 h on tire and metal, 6 h on cotton and hessian, and 3 h on wood. The effectiveness of disinfectants at inactivating this isolate was evaluated by adding them to sap from infected leaves which was then rubbed onto healthy tobacco plants. None of the plants became infected when bleach (42 g/liter sodium hypochlorite, diluted 1:4) or Virkon-S (potassium peroxymonosulfate 50% wt/wt, diluted to 1%) was used. A trace of infection remained after using nonfat milk powder (20% wt/vol). PVY infection sources were studied in 2011–2012 in the main potato growing regions of southwest Australia. In tests on >17,000 potato leaf samples, PVY was detected at low levels in seed (4/155) and ware (6/51) crops. It was also detected in volunteer potatoes from a site with a previous history of PVY infection in a seed crop. None of the 15 weed species tested were PVY infected. Plants of Solanum nigrum were symptomlessly infected with PVYO after sap inoculation, and no seed transmission was detected (>2,500 seeds). This study demonstrates PVYO can be transmitted by contact and highlights the need to include removal of volunteer potatoes and other on-farm hygiene practices (decontaminating tools, machinery, clothing, etc.) in integrated disease management strategies for PVY in potato crops.

scholarly journals Field Evaluation and Enzyme-Linked Immunosorbent Assay Detection of Potato Leaf Roll Virus, Potato Virus X and Potato Virus Y in Potato Germplasm

2017 ◽  
Vol 9 (7) ◽  
pp. 229 ◽  
Author(s):  
Romana Anjum ◽  
M. Aslam Khan ◽  
Kolawole Oluwaseun Olawale ◽  
Raheel Baber
Keyword(s):  
Immunosorbent Assay ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Rating Scale ◽  
Leaf Roll ◽  
Potato Virus X ◽  
Potato Leaf Roll Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potato Leaf ◽  
Potato Varieties

Polerovirus: potato leaf roll virus (PLRV), Potyvirus: potato virus Y (PVY) and Potexvirus: potato virus X (PVX) is more destructive and well distributed throughout the Pakistan. Incidence has been reported to be as high as 90%, 25%, and ≥ 15%, respectively in the potato growing regions. To find out the source of resistance, twenty-nine virus free potato varieties were grown under field conditions with good agricultural practices. The disease severity of PLRV, PVY and PVX was recorded to determine the level of resistance of the potato varieties according to the disease rating scale. Infectivity and biological assay of all twenty-nine varieties were done in green house on potato, Datura stramonium, Nicotiana glutinosa and Physalis floridana. Non-inoculated plants were served as control. Leaf samples from potato varieties were collected for serological detection of PLRV, PVY and PVX by Double antibody sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA). Out of twenty nine varieties, none of the variety was resistant to PLRV although three varieties; Mirrato, 394021-120 and Orla were moderately resistant. Only FD 48-4 and TPS 9813 showed resistance to PVX and PVY. While FD 3-10, FD 9616 and FD 37-13 were moderately to PVX and PVY. Rest of the varieties was found susceptible to all three viruses.

scholarly journals The effect of Ethrel on detection of potato virus Y (PVYº) in primarily infected potato plants

2013 ◽  
Vol 41 (2) ◽  
pp. 257-264
Author(s):  
Marian Staszewicz
Keyword(s):  
Statistical Analysis ◽  
Potato Virus ◽  
Active Ingredient ◽  
Potato Virus Y ◽  
Elisa Test ◽  
Field Conditions ◽  
Average Deviation ◽  
Potato Plants ◽  
Control Variant ◽  
And Control

The effect of ethylene (in the form of Ethrel) on the detection of PVYº in cv. Kaszubski potatoes primarily infected in field conditions, was studied. The plants were inoculated with PVYº and sprayed with Ethrel at a concentration of 100 mg/dm<sup>3</sup>, 10 cm<sup>3</sup> of solution were used per plant. The concentration of the active ingredient, CEPA, in this amount of solution was 1 mg/plant. The extinction value and PVYº detection were assessed in the leaf and tuber juices from Ethrel-treated plants and control variant. The ELISA test was used. The results were subjected to statistical analysis and the average deviation at <i>P</i> = 0.05 was calculated. It was shown that Ethrel lowered the value of extinction and decreased PVYº detection in the leaves and tubers of potato plants of the Kaszubski variety primarily infected in field conditions.

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