Purification of potato virus A and its detection in potato by enzyme-linked immunosorbent assay (ELISA)

1981 ◽  
Vol 58 (4) ◽  
pp. 181-189 ◽  
Author(s):  
R. P. Singh ◽  
J. G. McDonald
2019 ◽  
Vol 55 (4) ◽  
pp. 434-440
Author(s):  
N. A. Panferova ◽  
V. G. Panferov ◽  
I. V. Safenkova ◽  
Y. A. Varitsev ◽  
A. V. Zherdev ◽  
...  

Plant Disease ◽  
2004 ◽  
Vol 88 (10) ◽  
pp. 1160-1160 ◽  
Author(s):  
P. E. Thomas

Potato virus A (PVA; genus Potyvirus, family Potyviridae) occurs wherever potatoes are grown and may reduce tuber yields as much as 40%. Its host range consists of six experimental hosts (Lycopersicon pimpinellifolium (Jusl.) P. Mill., Nicandra physalodes (L.) Gaertn., Nicotiana tabacum L., Solanum demissum Lindl., S. demissum × S. tuberosum [2], and Nicotiana debneyi Domin.) and two natural hosts (S. tuberosum L. and S. betaceae (Cav.) Sendt.) (2). Aphids transmit PVA in a stylet-born manner. Its difficult mechanical transmission, caused by a low virus concentration in potato and other hosts (1), has constrained pathological research on the virus. In routine studies to identify virus isolates from the field, we discovered that N. megalosiphon Van Heurck & Mull. Agr. is a superior host of PVA that markedly facilitated diagnosis, selection for resistance to PVA, and other research applications. The efficiency of mechanical transmission of PVA to potato (5 duplicated assays and 10 plants per assay) ranged from 0 to 10% with PVA-infected potato as the virus source, 0 to 30% with Nicandra physalodes, 10 to 30% with N. tabacum cv. Samsun, and 20 to 80% with N. megalosiphon as the source of virus. The efficiency of mechanical transmission to four systemic hosts of PVA with potato (cv. Russet Burbank) as the source of virus (5 duplicated assays and 10 plants per assay) ranged from 0 to 20% to potato, 0 to 30% to Nicandra physalodes, 10 to 40% to N. tabacum cv. Samsun, and 80 to 100% to N. megalosiphon. The superiority of N. megalosiphon as a host and source of PVA was associated with a high virus concentration in tissues. Infected potato leaves yielded 0.32 to 0.54 mg of virus per kg of infected leaves, Nicandra physalodes yielded 0.37 to 0.66 mg per kg, N. tabacum cv. Samsun yielded 0.78 to 1.22 mg per kg, and N. megalosiphon yielded 5.16 to 9.39 mg per kg of infected leaves in five different purification experiments. These yields are based on the amount of virus isolated in sucrose gradients subjected to rate-zonal centrifugation as the last step in purification (3). The virus antigen concentrations of the original PVA-infected tissues measured using quantitative enzyme-linked immunosorbent assay ranked virus concentrations in the same relative order as purification but were nearly 2 times higher than were the purification yields. Similarly, local lesion assays on S. demissum A leaves (4) ranked infectious virus concentrations in the same order as did purification. Efficiency of aphid transmission from the four hosts was not assayed. Infected N. megalosiphon plants survived and served as sources of PVA for at least 1 year in a greenhouse. N. megalosiphon is an important new host of PVA because it facilitates the routine transmission of the virus and other manipulations essential for efficient research on control of the virus disease. References: (1) R. Bartels. No. 54 in: Descriptions of Plant Viruses. CMI, Kew, Surrey, UK, 1971. (2) A. Brunt. Page 77 in: Virus and Virus-Like Diseases of Potatoes and Production of Seed Potatoes. G Loebenstein et al., eds. Kluwer Academic Publishers, Dordrecht, the Netherlands, 2001. (3) P. E. Thomas and W. K. Kaniewski. Page 285 in: Virus and Virus-Like Diseases of Potatoes and Production of Seed Potatoes, G. Loebenstein et al., eds. Kluwer Academic Publishers, Dordrecht, the Netherlands, 2001. (4) R. E. Webb and R. W. Buck. Am Potato J. 32:248, 1955.


2017 ◽  
Vol 9 (7) ◽  
pp. 229 ◽  
Author(s):  
Romana Anjum ◽  
M. Aslam Khan ◽  
Kolawole Oluwaseun Olawale ◽  
Raheel Baber

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.


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