scholarly journals Purification of Potato Virus Y for Polyclonal Antisera Production and Assessment of Antisera Specificity through DAS-ELISA

Author(s):  
Ranima Mishra ◽  
P.D. Nath ◽  
B. Raigond ◽  
R.C. Boro ◽  
Shankar Hemanta Gogoi ◽  
...  
2004 ◽  
Vol 22 (3) ◽  
pp. 521-524 ◽  
Author(s):  
Julio Daniels ◽  
Arione da S. Pereira

O vírus do enrolamento da folha da batata (Potato leafroll virus, PLRV) e o vírus Y da batata (Potato virus Y, PVY) constituem as principais causas da degenerescência da batata-semente no Brasil. Com o objetivo de determinar, nas condições do Rio Grande do Sul, a resistência de campo de genótipos de batata à infecção por estes vírus, avaliaram-se, na presença de infectores, durante três plantios consecutivos de primavera, 20 cultivares e clones de batata. A detecção dos vírus foi efetuada por meio de testes sorológicos (DAS-ELISA). Pela análise de agrupamento os genótipos foram separados em três grupos para resistência ao PLRV (Elvira, Achat, Bintje, Monalisa, Monte Bonito, Panda e Araucária, resistentes; Baronesa, Asterix, Atlantic, 2CRI-1149-1-78, C-1226-35-80, Astrid, C-1714-7-94, A-1139-12-92, Macaca, Eliza e Santo Amor, suscetíveis; Catucha e Cristal, muito suscetíveis) e em quatro grupos para resistência ao PVY (Asterix, Astrid, Catucha, Cristal, Macaca, Monte Bonito, A-1139-12-92, C-1226-35-80 e C-1714-7-94, resistentes; Baronesa, Santo Amor, Monalisa, Panda e 2CRI-1149-1-78, resistentes intermediários; Bintje, Atlantic, Elvira e Araucária, suscetíveis; Achat e Eliza, muito suscetíveis).


2018 ◽  
Vol 3 (2) ◽  
pp. 381-390
Author(s):  
Rodolfo Velásquez-Valle ◽  
Luis Roberto Reveles-Torres ◽  
Jaime Mena-Covarrubias

A nivel mundial el cultivo de chile es afectado por más de 60 enfermedades virales; sin embargo, poco se conoce acerca de ellas en el área productora de chile seco del norte centro de México por lo que el objetivo del presente trabajo consistió en detectar la presencia y sintomatología de cinco virus en parcelas comerciales de chile seco en los estados mencionados. Plantas de chile de los tipos mirasol y ancho fueron muestreadas y se anotó la presencia de síntomas como enanismo, clorosis, deformación de hojas, defoliación, necrosis vascular y ramas unidas. Las muestras fueron analizadas mediante la técnica DAS- ELISA empleando los antisueros para el virus del mosaico del tabaco (Tobacco mosaic virus: TMV), mosaico del pepino (Cucumber mosaic virus: CMV), Y de la papa (Potato virus Y: PVY), moteado del chile (Pepper mottle virus: PepMoV) y jaspeado del tabaco (Tobacco etch virus: TEV). Esos virus fueron identificados en plantas de chile colectadas en las parcelas comerciales de chile seco de los tres estados antes mencionados.


2002 ◽  
Vol 20 (3) ◽  
pp. 510-513 ◽  
Author(s):  
Julio Daniels ◽  
Antônio Carlos F. Silva ◽  
Zilmar S. Souza ◽  
Jurema Schons

A infecção pelo vírus do enrolamento da folha da batata (Potato leafroll virus, PLRV) e pelo vírus Y da batata (Potato virus Y, PVY) constitui-se na principal causa da degenerescência da batata-semente no Brasil. Durante dois períodos de cultivo, setembro a dezembro de 1999 e março a junho de 2000, avaliou-se o percentual de infeção de batata-semente básica pelo PLRV e pelo PVY, em regiões produtoras de batata do Rio Grande do Sul e de Santa Catarina. No primeiro período foram avaliadas sorologicamente (DAS-ELISA) 623 amostras de nove regiões, observando-se percentagens de infeção de 17% para PVY e de 3% para PLRV. A percentagem de infeção por PVY e PLRV em cada cultivar foi, respectivamente, Baronesa 7,5 e 0,3%, Catucha 22 e 19%, Elvira 42 e 0% e Monalisa 16 e 4%. No plantio do segundo período usou-se parte dos tubérculos colhidos no primeiro, e foram avaliadas 301 amostras de duas regiões, constatando-se uma percentagem média de infeção de 58% para o PVY e de 11% para PLRV. A percentagem, por cultivar, foi Baronesa 67 e 6%, Catucha 68 e 49%, Elvira 94 e 2% e Monalisa 35 e 0%.


2009 ◽  
Vol 44 (9) ◽  
pp. 1127-1130 ◽  
Author(s):  
André Nepomuceno Dusi ◽  
Cristiane Lopes de Oliveira ◽  
Paulo Eduardo de Melo ◽  
Antonio Carlos Torres

The objective of this work was to evaluate the resistance of genetically modified clones of potato to Potato virus Y (PVY) under field conditions. Genetically modified plants were compared with nontransformed plants of the same cultivar. The plots were flanked with potato plants infected with both PVYº and PVY N strains (spread lines), in order to provide the experimental area with the source of virus, which was naturally spread by the native aphid population. The experiment was weekly monitored by visual inspections and by DAS-Elisa in the plants produced from the harvested tubers, in order to evaluate the resistance of transgenic plants throughout the plant growth cycle. By the end of the third year, no infection symptoms were observed in the 1P clone; clone 63P showed 1% of infection, in contrast to about 90% of nontransformed plants infected. The stable expression of resistance to PVY provided by the coat protein gene was obtained in genetically modified clones of potato plants cultivar Achat under field conditions, during three consecutive years.


Plant Disease ◽  
2008 ◽  
Vol 92 (2) ◽  
pp. 312-312 ◽  
Author(s):  
L. Cardin ◽  
B. Moury

Nicotiana mutabilis Stehmann & Semir is a recently described perennial plant species from southern Brazil that produces long floral stems with white to deep pink flowers and is used for its ornamental quality. In 2003, leaf mosaic symptoms were observed in all 30 N. mutabilis plants in a nursery in the south of France. Observation of crude sap preparations with the electron microscope revealed numerous flexuous particles, 700 to 730 nm long and approximately 11 nm wide, associated with “pinwheel”-like cytoplasmic inclusions, typical of the family Potyviridae. A range of plant species inoculated with extracts from five of the symptomatic plants showed reactions typical of Potato virus Y (PVY) (2), and the presence of the virus was confirmed by positive reactions in double-antibody sandwich (DAS)-ELISA with polyclonal antibodies raised against PVY. To test if PVY was responsible for the symptoms observed in N. mutabilis, an isolate was multiplied in N. tabacum cv. Xanthi plants after isolation from local lesions on Chenopodium amaranticolor and was then mechanically inoculated to 12 seedlings of N. mutabilis cv. Marshmallow. After 3 weeks, the 12 inoculated plants showed systemic vein clearing symptoms and PVY was detected by DAS-ELISA. Reverse transcription (RT)-PCR tests using PVY-polyvalent primers (5′-GATGGTTGCCTTGGATGATG and 5′-TAAAAGTAGTACAGGAAAAGCCA) covering the coat protein (CP) coding region amplified a single DNA fragment of the expected 900 bp from total RNA extracts from Xanthi plants inoculated with the five isolates. One of these DNA products was directly sequenced (GenBank Accession No. EU252529) and several accepted methods of phylogenetic analysis compared this sequence to 80 available PVY CP coding sequences and showed that the N. mutabilis PVY belonged to the C1 group (1). Similar to the other PVY strains in the C group, the N. mutabilis isolate was able to induce hypersensitive local lesions in leaves of potato genotypes carrying the Nc gene. However, contrary to the other characterized C1 isolates (1), it was unable to infect systemically cv. Yolo Wonder pepper plants. That peculiar behavior makes the N. mutabilis isolate a tool to identify the viral determinants controlling the host range of PVY. References: (1) B. Blanco-Urgoiti et al. J. Gen. Virol. 79:2037, 1998. (2) C. Kerlan. No. 414 in: Descriptions of Plant Viruses. CMI/AAB, Kew, Surrey, UK, 2006.


1994 ◽  
Vol 3 (2) ◽  
pp. 207-211
Author(s):  
Maija Vihanen-Rantanen ◽  
Reijo Sironen ◽  
Matti Vuento

Synthetic, overlapping peptides representing the entire amino acid sequence of potato virus Y (PVY) coat protein were used to affinity-purify antibodies from polyclonal antisera to PVY. In testing the binding of the purified antibodies to PVY particles, antigenic epitopes were identified. The N-terminal and C-terminal regions of the PVY coat protein were found to contain most of the antigenic epitopes. The results will facilitate the development of detection methods for PVY based on synthetic peptides.


Agronomy ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1617
Author(s):  
Anna Depta ◽  
Teresa Doroszewska ◽  
Anna Czubacka

Nicotiana mutabilis is a recently discovered species within the genus Nicotiana. The aim of the present study was to evaluate its resistance to Potato virus Y (PVY) and Tomato spotted wilt virus (TSWV). Molecular analysis was performed to detect the Va gene determining susceptibility to PVY and the SCAR marker associated with resistance to TSWV. Resistance tests were carried out under greenhouse conditions through artificial inoculation with one TSWV and two PVY isolates. In order to confirm the presence of the viruses in plants, DAS-ELISA tests were performed using antibodies against PVY and TSWV. The results indicated the absence of the PVY susceptibility gene and the presence of the TSWV resistance gene in the genome of N. mutabilis. This species was considered tolerant to the two PVY isolates tested because, despite the positive DAS-ELISA results, the infected plants showed vein clearing and chlorotic spots but no vein necrosis. As a result of TSWV inoculation, N. mutabilis showed a hypersensitive response; however, after four months, 30% of the inoculated plants showed systemic infection. This species extends the genetic variation in the genus Nicotiana and, because of its tolerance to PVY and partial resistance to TSWV, it may be a potential source of resistance to these viruses.


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.


Bragantia ◽  
2008 ◽  
Vol 67 (2) ◽  
pp. 391-399 ◽  
Author(s):  
Silvia Regina Luz Palazzo ◽  
Addolorata Colariccio ◽  
Arlete Marchi Tavares de Melo

Amostras de tomateiro (Lycopersicon esculentum Mill) ‘Alambra’ coletadas nas regiões produtoras de Elias Fausto, Monte-Mor e Mogi-Guaçú (SP), com sintomas de amarelecimento foliar generalizado semelhante aqueles causados por vírus foram submetidas à identificação do agente causal, por testes biológicos de transmissão mecânica, pela determinação do círculo de hospedeiras, empregando-se plantas indicadoras e diferenciadoras pertencentes às famílias Chenopodiaceae e Solanaceae pela identificação sorológica por PTA-ELISA do Potato virus Y (PVY), Pepper yellow mosaic virus (PepYMV), Cucumber mosaic virus (CMV), Tomato mosaic virus (ToMV) e DAS-ELISA com antissoros policlonais para as espécies Tomato spot wilt vírus (TSWV), Tomato chlorotic spot virus (TCSV), Groundnut ringspot virus (GRSV), Chrysanthemum stem necrosis virus (CSNV) e anticorpos monoclonais para as estirpes do PVY comum (PVYº), PVY necrótico (PVY N) e PVY clorótico (PVY C). Das amostras coletadas 19 reagiram positivamente, com o PVY em PTA-ELISA e PVYºem DAS-ELISA. Plantas de Chenopodium amaranticolor reagiram com sintoma local e plantas de Nicotiana glutinosa, N. tabacum ‘WB’, N. sylvestris, N.debneyi, N. tabacum ‘Sansun’ reagiram com sintomas de mosaico sistêmico; tomateiros ‘Alambra’ manifestaram sintomas de mosaico-amarelo. Plantas de Datura stramonium, D. metel e C. annuum ‘Magda’ não foram infectadas. A ausência de sintomas em C.annuum ‘Magda’, identificou a presença do PVY patotipo 1 (PVYº1), nas 19 amostras. Como foi identificada a mesma espécie de vírus nas amostras, optou-se pela inoculação do isolado de tomate ‘Alambra’ de Elias Fausto nos dezenove acessos de Lycopersicon spp. do Banco Ativo de Germoplasma (BAG)- IAC. O delineamento dos experimentos foi inteiramente casualizado. A reação dos acessos foi avaliada pela manifestação dos sintomas, pelos resultados positivos ou negativos após testes de retro-inoculação e PTA-ELISA, pela análise do χ2 utilizando-se a proporção de plantas sintomáticas e assintomáticas, com resultados positivos ou que não manifestaram sintomas e reagiram negativamente, em PTA-ELISA, constituindo, portanto fontes potenciais de genes de resistência para o PVY em tomate.


Sign in / Sign up

Export Citation Format

Share Document