Development of Enzyme-Linked Immunosorbent Assay with Tiramine Amplification for the Detection of Potato Virus X

2019 ◽  
Vol 55 (4) ◽  
pp. 434-440
Author(s):  
N. A. Panferova ◽  
V. G. Panferov ◽  
I. V. Safenkova ◽  
Y. A. Varitsev ◽  
A. V. Zherdev ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Potato Virus ◽  
Potato Virus X ◽  
Enzyme Linked Immunosorbent Assay

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 Epidemic of Potato virus Y and Cucumber mosaic virus in Henan Province Tobacco

Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 447-447 ◽  
Author(s):  
X. D. Li ◽  
Y. Q. Li ◽  
H. G. Wang
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Early Stage ◽  
Potato Virus X ◽  
Henan Province ◽  
Enzyme Linked Immunosorbent Assay ◽  
Resistant Varieties ◽  
Vein Necrosis

Flue-cured tobacco is an important crop in Henan Province, China. During the 2000 growing season, many tobacco plants showed various degrees of mottling, mosaic, vein clearing, or vein necrosis in most of the counties. Some plants even died at an early stage of growth. A survey was conducted in May-June in several tobacco-growing counties, and the incidence of symptomatic plants in individual fields ranged from 10 to 85%. The most widely planted tobacco varieties, NC89, K326, and K346, were highly susceptible. Symptomatic plants were collected from Jiaxian and Xiangcheng counties and samples were tested by enzyme-linked immunosorbent assay for Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Potato virus X (PVX). Of 65 samples tested, 21 were positive for only PVY, 16 positive for only CMV, one each was positive for only TMV or PVX. Nineteen samples were doubly infected with various combinations of these viruses and six were infected with combinations of three viruses. The causal agent(s) in the remaining sample could not be determined. In total, CMV was detected in 40 samples, PVY in 38, PVX in 10, and TMV in 7 samples. TMV and CMV used to be the most important viruses and PVY occurred only rarely. But PVY has become prevalent in Henan and in neighboring Shandong province (2). CMV and TMV were reported to be the most prevalent viruses in Shanxi (1) and Fujian Provinces (3). Because resistant varieties are not available, and mixed infections are more common, the results presented here explain why huge damage is occurring in tobacco crops in recent years. Some varieties are partially resistant to TMV and CMV but the varieties commonly grown are highly susceptible to PVY. Therefore, breeding for resistance to viruses, especially to PVY, is urgent to control the occurrence of tobacco viral diseases. References: (1) J. L. Cheng et al. Acta Tabacaria Sin. 4:43, 1998. (2) J. B. Wang et al. Chinese Tobacco Sci. 1:26, 1998. (3) L. H. Xie et al. Acta Tabacaria Sin. 2:25, 1994.

scholarly journals Natural Occurrence of Viruses in Lycopersicon spp. in Ecuador

Plant Disease ◽  
2005 ◽  
Vol 89 (11) ◽  
pp. 1244-1244 ◽  
Author(s):  
S. Soler ◽  
C. López ◽  
F. Nuez
Keyword(s):  
Mosaic Virus ◽  
Potato Virus ◽  
Potato Virus X ◽  
Tomato Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Natural Occurrence ◽  
Single Plant ◽  
Rt Pcr ◽  
Specific Primers ◽  
In The Wild

The Andean region is home of important genetic diversity for the genus Lycopersicon. A survey of three asymptomatic populations of L. hirsutum, 17 of L. parviflorum, 188 of L. pimpinellifolium, and four cultivated populations of L. esculentum was made in nine departments of Ecuador. Samples were analyzed serologically for Tomato spotted wilt virus (TSWV), Tomato mosaic virus (ToMV), Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), Potato virus Y (PVY), Potato virus X (PVX), Groundnut ringspot virus (GRSV), Tomato chlorosis spot virus (TCSV), and Pepino mosaic virus (PepMV). Samples positive as determined using double-antibody sandwich enzyme-linked immunosorbent assay (absorbance values three times higher than negative controls) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR) with virus-specific primers. L pimpinellifolium was the only species of the four found to be infected with viruses. In the department of Manabí, ToMV was detected in 15 of 16 plants from one population, but only a single plant was infected with PepMV. In this department, PepMV was also detected in a single-plant population that corresponded to a volunteer plant found in the wild and TSWV was detected in another plant. In Esmeraldas and Guayas, two single-plant populations were found infected with PepMV and CMV, respectively. TMV, PVY, PVX, GRSV, and TCSV were not detected in this survey. Specific primers were selected for ToMV (To1/To2, genome coordinates 3498-3518/4902-4922, AJ417701), PepMV (Pe1/Pe2 genome coordinates 5030-5050/5913-5935, AJ606359), CMV (Cm1/Cm2 genome coordinates 541-561/1756-1779, D00356), and TSWV (Ts1/Ts2 genome coordinates 4078-4101/4738-4769, AF208498). Amplicons of the expected size were obtained using RT-PCR and then cloned and sequenced. DNA fragments of ToMV, PepMV, and TSWV showed identities greater than 99% with respective sequences in the GenBank database. The highest identity of the CMV DNA fragment was 92% with an isolate from Indonesia (AB042292). The occurrence of viruses such as CMV, ToMV, and TSWV in coastal Ecuador was not surprising. However, infected plants were not found among the samples collected in the departments of Azuay, Carchí, El Oro, Imbabura, Loja, and Pichincha in eastern Ecuador. L. chilense, L. chmielewskii, L. parviflorum, and L. peruvianum were previously reported as natural hosts of PepMV in central and southern Peru (2), and the virus was also detected in L. esculentum in Chile (1). Our results show that PepMV now occurs in wild L. pimpinellifolium populations along the Pacific coast of the South American continent and that it must have efficient means of transmission, although no specific vectors have as yet been identified for this virus. To our knowledge, this is the first report of PepMV in Ecuador and L. pimpinellifolium as a natural host of PepMV. References: (1) M. Muñoz et al. Fitopatología 37:67, 2002. (2) S. Soler et al. J. Phytopathol. 150:49, 2002.

scholarly journals Presence and distribution of economically important potato viruses in Montenegro

2011 ◽  
Vol 26 (2) ◽  
pp. 117-127
Author(s):  
Jelena Zindovic
Keyword(s):  
Potato Virus ◽  
Leaf Roll ◽  
Potato Virus X ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potato Leaf ◽  
Potato Virus S ◽  
Potato Varieties ◽  
Potato Viruses ◽  
Das Elisa

The research was carried out, in the period 2002-2004 in order to determine the presence and distribution of potato viruses at 12 different locations and on 9 different potato varieties grown in Montenegro. The research included collecting of samples in seed potato crops and testing of six economically important potato viruses: Potato leaf roll virus (PLRV), Potato virus Y (PVY), Potato virus X (PVX), Potato virus S (PVS), Potato virus A (PVA) i Potato virus M (PVM). Using the direct enzyme-linked immunosorbent assay (DAS-ELISA) and commercial antisera specific for six potato viruses, it was found that PVY was the most frequent virus during the three-year research period. The second frequent virus was PVS, followed by PVA, PLRV, PVM and PVX. Single and mixed infections were detected, and the most prevalent were the single infections of PVY. Also, in the period 2002-2004, PVY had the highest distribution and the number of present viruses was different at different localities and on different potato varieties. Further investigations were related to detailed characterization of the most prevalent virus (PVY), which is at the same time economically the most important one. Serological characterization of PVY was performed utilizing DAS-ELISA kit with commercial monoclonal antibodies specific for detection of the three strain groups of PVY, and the two strain groups - necrotic (PVYN/PVYNTN) and common (PVYO), were identified. Necrotic strains were prevalent in 2002 and 2004, while in 2003 PVYO was the most frequent strain in virus population. The presence of stipple streak strain (PVYC) was not detected in any of the tested samples.

scholarly journals Macroarray Detection of Eleven Potato-Infecting Viruses and Potato spindle tuber viroid

Plant Disease ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 730-740 ◽  
Author(s):  
Bright Agindotan ◽  
Keith L. Perry
Keyword(s):  
Mosaic Virus ◽  
Potato Virus ◽  
Tobacco Rattle Virus ◽  
Alfalfa Mosaic Virus ◽  
Potato Virus X ◽  
Potato Spindle Tuber Viroid ◽  
Apparent Competition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potato Seed ◽  
Potato Virus S

A macroarray was developed for the detection of 11 potato viruses and Potato spindle tuber viroid. The 11 viruses detected included those commonly found or tested for in North American potato seed certification programs: Alfalfa mosaic virus, Cucumber mosaic virus, Potato mop top virus, Potato leafroll virus, Potato latent virus, Potato virus A, Potato virus M, Potato virus S, Potato virus X, Potato virus Y, and Tobacco rattle virus. These viruses were detected using oligonucleotide 70-mer probes and labeled targets prepared by a random primed amplification procedure. Potato plants analyzed included those infected with 12 reference virus stocks and 36 field isolates. Results from the macroarray were entirely consistent with those obtained using a standard serological assay (enzyme-linked immunosorbent assay). Four isolates of Potato spindle tuber viroid, in mixed infection with one or more viruses, also were detected in the array, although strong hybridization signals required amplification with viroid-specific primers in combination with anchored-random primers. In individual plants, up to four viruses, or a viroid plus two viruses, were detected, with no apparent competition or inhibition. Macroarrays are a cost-effective approach to the simultaneous diagnostic detection of multiple pathogens from infected plants.

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