scholarly journals First Report of Tomato spotted wilt virus in Peppers and Tomato in the Dominican Republic

Plant Disease ◽  
2014 ◽  
Vol 98 (1) ◽  
pp. 163-163 ◽  
Author(s):  
R. T. Martínez ◽  
S. Poojari ◽  
S. A. Tolin ◽  
X. Cayetano ◽  
R. A. Naidu
Keyword(s):  
Dominican Republic ◽  
Tomato Spotted Wilt Virus ◽  
Caribbean Region ◽  
Rt Pcr ◽  
Spot Virus ◽  
Tomato Spotted Wilt ◽  
Fruit Samples ◽  
The Caribbean ◽  
Wilt Virus ◽  
Pepper Plants

In the Dominican Republic, green bell pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.) are widely cultivated under protected greenhouse conditions as high value commercial crops for export. For the past 2 to 3 years, pepper and tomato have been observed in protected crop facilities in Jarabacoa and Constanza in the North Region with chlorotic and necrotic spots and rings on leaves, petioles, and stems, leaf bronzing, and tip necrosis. Fruits on symptomatic pepper and tomato plants showed concentric rings, irregular chlorotic blotches and deformation, and uneven maturation and development. Incidence on pepper and tomato was 20 to 100% and 5 to 20%, respectively. In initial tests, leaves and fruits from each of 20 symptomatic tomato and pepper plants from several greenhouse facilities were reactive in Tomato spotted wilt virus (TSWV; genus Tospovirus, family Bunyaviridae) immunostrip assays (Agdia, Inc., Elkhart, IN). Since these immunostrips are known to react with other tospoviruses, such as Tomato chlorotic spot virus (TCSV) and Groundnut ring spot virus, additional molecular diagnostic assays were conducted. Leaf and fruit samples from symptomatic plants were imprinted on nitrocellulose membrane (NCM) (2), air-dried, and sent to Washington State University for confirmatory tests. Viral nucleic acids were eluted from NCM discs (1) and subjected to reverse transcription (RT)-PCR using primers gL3637 (CCTTTAACAGTDGAAACAT) and gL4435 (CATDGCRCAAGARTGRTARACAGA) designed to amplify a portion of the L RNA segment of several tospoviruses (3). A single DNA product of ~800 bp was amplified from all samples. Amplicons from two tomato (leaf and fruit) and one pepper fruit samples were cloned separately into pCR2.1 (Invitrogen Corp., Carlsbad, CA). Two independent clones per amplicon were sequenced in both orientations. Sequence analyses of these clones (GenBank Accession Nos. KF 219673 to 75) showed 100% nucleotide sequence identity among themselves and 97% identity with corresponding L RNA sequences of pepper isolates of TSWV from Taiwan (HM180088) and South Korea (HM581940), 94 to 95% with tomato isolates of TSWV from South Korea (HM581934) and Hawaii (AY070218), and 89% with a tomato isolate from Indonesia (FJ177301). These results further confirm the presence of TSWV in symptomatic tomato and pepper plants. A comparison of TSWV sequences from the Dominican Republic with TSWV isolates from the United States and other countries in the Caribbean region could not be made due to the absence of corresponding sequences of the L-RNA of the virus from these countries in GenBank. TSWV-positive samples were negative for TCSV in RT-PCR, indicating the absence of this tospovirus that has been reported in the Caribbean region (data not shown). To our knowledge, this is the first confirmed report of TSWV in tomatoes and peppers in the Dominican Republic. The presence of vector thrips, Frankliniella occidentalis, on symptomatic plants was also confirmed, suggesting a role in the spread of TSWV under greenhouse conditions. Recent surveys identified some greenhouses with 100% symptomatic peppers. The presence of TSWV in tomato and pepper has important implications for the domestic and export vegetable industry in the Dominican Republic because of the broad host range of the virus (4). It is critical for commercial producers to monitor TSWV and deploy appropriate management strategies to limit virus spread. References: (1) O. J. Alabi et al. J. Virol. Methods 154:111, 2008. (2) P.-G. S. Chang et al. J. Virol. Methods 171:345, 2011. (3) F. H. Chu et al. Phytopathology 91:361, 2001. (4) G. Parrella et al. J. Plant Pathol. 85:227, 2003.

scholarly journals Caracterização do Tomato chlorotic spot virus isolado de jiló no Vale do Paraíba, Estado de São Paulo

2002 ◽  
Vol 27 (3) ◽  
pp. 285-291 ◽  
Author(s):  
MARCELO EIRAS ◽  
ALEXANDRE L. R. CHAVES ◽  
ADDOLORATA COLARICCIO ◽  
RICARDO HARAKAVA ◽  
JANSEN DE ARAUJO ◽  
...  
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Sao Paulo ◽  
Ringspot Virus ◽  
São Paulo ◽  
Rt Pcr ◽  
Spot Virus ◽  
Chlorotic Spot ◽  
Tomato Spotted Wilt ◽  
Wilt Virus ◽  
Das Elisa

Os tospovírus são responsáveis por perdas significativas em diversas culturas, principalmente solanáceas. No município de São José dos Campos (SP), plantas de jiló (Solanum gilo) apresentando sintomas de mosaico, bolhosidades, nanismo e queda acentuada da produção foram coletadas para análise. Visando a caracterização do agente causador dos sintomas, testes biológicos, elétrono microscópicos, sorológicos e moleculares foram realizados. Através de inoculação mecânica em plantas indicadoras das famílias Amaranthaceae, Chenopodiaceae e Solanaceae obtiveram-se resultados típicos aos esperados para tospovírus. Ao microscópio eletrônico de transmissão, observaram-se, em contrastação negativa, partículas pleomórficas com diâmetro entre 80 e 110 nm e em cortes ultra-finos partículas presentes em vesículas do retículo endoplasmático. Através de DAS-ELISA, identificou-se o Tomato chlorotic spot virus (TCSV). A partir de RNA total extraído de folhas infetadas, amplificaram-se, via RT-PCR, fragmentos correspondentes ao gene da proteína do capsídeo (cp) os quais foram seqüenciados e comparados com outros depositados no "GenBank". A homologia de nucleotídeos e aminoácidos deduzidos foi respectivamente de 99 e 95% quando comparada com seqüências de isolados de TCSV. A comparação com as outras espécies do gênero Tospovirus apresentou valores de homologia entre 72 e 84%. Estes resultados confirmam a identidade deste vírus como pertencente à espécie TCSV, que é predominante no Estado de São Paulo e importante patógeno de outras plantas cultivadas. Além disso, variedades de jiló quando inoculadas foram susceptíveis tanto ao TCSV como às espécies Tomato spotted wilt virus (TSWV) e Groundnut ringspot virus (GRSV).

scholarly journals First Report of Vidalia Onion (Allium cepa) Naturally Infected with Tomato spotted wilt virus and Iris yellow spot virus (Family Bunyaviridae, Genus Tospovirus) in Georgia

Plant Disease ◽  
2004 ◽  
Vol 88 (11) ◽  
pp. 1285-1285 ◽  
Author(s):  
S. W. Mullis ◽  
D. B. Langston ◽  
R. D. Gitaitis ◽  
J. L. Sherwood ◽  
A. C. Csinos ◽  
...  
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Thrips Tabaci ◽  
Iris Yellow Spot Virus ◽  
Yellow Spot ◽  
Unknown Etiology ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Spot Virus ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

Vidalia onion is an important crop in Georgia's agriculture with worldwide recognition as a specialty vegetable. Vidalia onions are shortday, Granex-type sweet onions grown within a specific area of southeastern Georgia. Tomato spotted wilt virus (TSWV) has been endemic to Georgia crops for the past decade, but has gone undetected in Vidalia onions. Tobacco thrips (Frankliniella fusca) and Western flower thrips (Frankliniella occidentalis) are the primary vectors for TSWV in this region, and a number of plant species serve as reproductive reservoirs for the vector or virus. Iris yellow spot virus (IYSV), an emerging tospovirus that is potentially a devastating pathogen of onion, has been reported in many locations in the western United States (2,4). Thrips tabaci is the known vector for IYSV, but it is unknown if noncrop plants play a role in its epidemiology in Georgia. During October 2003, a small (n = 12) sampling of onions with chlorosis and dieback of unknown etiology from the Vidalia region was screened for a variety of viruses, and TSWV and IYSV infections were serologically detected. Since that time, leaf and bulb tissues from 4,424 onion samples were screened for TSWV and IYSV using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) with commercial kits (Agdia Inc., Elkhart, IN). Samples were collected from 53 locations in the Vidalia region during the growing season between November 2003 and March 2004. Plants exhibiting stress, such as tip dieback, necrotic lesions, chlorosis or environmental damage were selected. Of these, 306 were positive for TSWV and 396 were positive for IYSV using positive threshold absorbance of three times the average plus two standard deviations of healthy negative onion controls. Positive serological findings of the onion tissues were verified by immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) for TSWV (3) and RT-PCR for IYSV (1). In both instances, a region of the viral nucleocapsid (N) gene was amplified. The PCR products were analyzed with gel electrophoresis with an ethidium bromide stain in 0.8% agarose. Eighty-six percent (n = 263) of the TSWV ELISA-positive samples exhibited the expected 774-bp product and 55 percent (n = 217) of the IYSV ELISA-positive samples exhibited the expected 962-bp product. The reduced success of the IYSV verification could be attributed to the age and deteriorated condition of the samples at the time of amplification. Thrips tabaci were obtained from onion seedbeds and cull piles within the early sampling (n = 84) and screened for TSWV by the use of an indirect-ELISA to the nonstructural (NSs) protein of TSWV. Of the thrips sampled, 25 were positive in ELISA. While the incidence of IYSV and TSWV in the Vidalia onion crop has been documented, more research is needed to illuminate their potential danger to Vidalia onions. References: (1) I. Cortês et al. Phytopathology 88:1276, 1998. (2) L. J. du Toit et al. Plant Dis. 88:222, 2004. (3) R. K. Jain et al. Plant Dis. 82:900, 1998. (4) J. W. Moyer et al. (Abstr.) Phytopathology 93(suppl.):S115, 2003.

scholarly journals İstanbul İli Adalar İlçesi’nde Hobi Bahçeleri ve Peyzaj Alanlarında Yetiştirilen Süs Bitkilerinde Tospovirüslerin Saptanması

2017 ◽  
Vol 5 (9) ◽  
pp. 1119
Author(s):  
Fatma Şafak ◽  
Muharrem Arap Kamberoğlu
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Ornamental Plants ◽  
Necrotic Spot ◽  
Rt Pcr ◽  
Spot Virus ◽  
Tomato Spotted Wilt ◽  
Hydrangea Macrophylla ◽  
Wilt Virus ◽  
Pittosporum Tobira ◽  
Das Elisa

This study was conducted in order to detect Tomato spotted wilt virus (TSWV), Irish yellow spot virus (IYSV) and Impatients necrotic spot virus (INSV) in ornamental plants growing in hobby gardens and landscaping areas in Adalar district (Büyükada, Heybeliada, Kınalıada and Burgazada) of Istanbul province between 2015 and 2016. During the surveys carried out in that district, the samples were collected from both simptomatologically suspicious ornamental plants and the plants which did not show any symptoms. All of the collected samples (n=150) were firstly tested by Double Antibody Sandwich (DAS) ELISA and none of the samples were found to be infected with TSWV and INSV. The samples detected to be positive with IYSV by ELISA tests were then used in RT-PCR studies. At the RT-PCR using the IYSV-465c; IYSV-239f primer pair, a band with a size of 240 bp was observed for Pittosporum tobira and Hydrangea macrophylla. Therefore, the presence of IYSV infection in Adalar was also confirmed molecularly.

Comparison of ELISA and RT-PCR assays for the detection of Tomato spotted wilt virus in peanut

2009 ◽  
Vol 36 (2) ◽  
pp. 133-137 ◽  
Author(s):  
P. M. Dang ◽  
D. L. Rowland ◽  
W. H. Faircloth
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Rates ◽  
Rt Pcr ◽  
Chi Square ◽  
Tomato Spotted Wilt ◽  
Significant Difference ◽  
Pcr Assays ◽  
Chi Square Analysis ◽  
Wilt Virus

Abstract Diagnosis of Tomato spotted wilt virus (TSWV) in peanut can be accomplished by enzyme-linked immunosorbent assay (ELISA) or reverse transcription polymerase chain reaction (RT-PCR) but there has been no report of a direct comparison of the success of the two assays in evaluating infection rates of field-grown peanut. We collected peanut root samples from field-grown plants, 76 in 2006 and 48 in 2007, and tested these samples by both ELISA and RT-PCR assays for the presence of TSWV. Out of 124 samples, 50 (40.3%) and 57 (46.0%) were positive for TSWV by ELISA and RT-PCR respectively. In 13.7% of these samples, ELISA and RT-PCR differed in their results. However, Chi square analysis showed no significant difference between the results for these two assays. This result supports the conclusion that ELISA and RT-PCR are comparable for detecting TSWV infection rates in field-grown peanuts.

Real-time RT-PCR fluorescent detection of tomato spotted wilt virus

2000 ◽  
Vol 88 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Cassie A Roberts ◽  
Ralf G Dietzgen ◽  
Lisa A Heelan ◽  
Donald J Maclean
Keyword(s):  
Real Time ◽  
Tomato Spotted Wilt Virus ◽  
Fluorescent Detection ◽  
Rt Pcr ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

scholarly journals First Report of Tomato spotted wilt virus in Blackberry Lily in North America

Plant Disease ◽  
2003 ◽  
Vol 87 (1) ◽  
pp. 102-102 ◽  
Author(s):  
S. Adkins ◽  
L. Breman ◽  
C. A. Baker ◽  
S. Wilson
Keyword(s):  
North America ◽  
Tomato Spotted Wilt Virus ◽  
Amino Acid Sequences ◽  
South Florida ◽  
N Gene ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

Blackberry lily (Belamcanda chinensis (L.) DC.) is an herbaceous perennial in the Iridaceae characterized by purple-spotted orange flowers followed by persistent clusters of black fruit. In July 2002, virus-like symptoms including chlorotic ringspots and ring patterns were observed on blackberry lily leaves on 2 of 10 plants in a south Florida ornamental demonstration garden. Inclusion body morphology suggested the presence of a Tospovirus. Tomato spotted wilt virus (TSWV) was specifically identified by serological testing using enzyme-linked immunosorbent assay (Agdia, Elkhart, IN). Sequence analysis of a nucleocapsid (N) protein gene fragment amplified by reverse transcription-polymerase chain reaction (RT-PCR) with primers TSWV723 and TSWV722 (1) from total RNA confirmed the diagnosis. Nucleotide and deduced amino acid sequences of a 579 base pair region of the RT-PCR product were 95 to 99% and 95 to 100% identical, respectively, to TSWV N-gene sequences in GenBank. Since these 2-year-old plants were grown on-site from seed, they were likely inoculated by thrips from a nearby source. Together with a previous observation of TSWV in north Florida nursery stock (L. Breman, unpublished), this represents, to our knowledge, the first report of TSWV infection of blackberry lily in North America although TSWV was observed in plants of this species in Japan 25 years ago (2). References: (1) S. Adkins, and E. N. Rosskopf. Plant Dis. 86:1310, 2002. (2) T. Yamamoto and K.-I. Ohata. Bull. Shikoku Agric. Exp. Stn. 30:39, 1977.

scholarly journals Impatiens necrotic spot virus in Greenhouse-Grown Potatoes in New York State

Plant Disease ◽  
2005 ◽  
Vol 89 (3) ◽  
pp. 340-340 ◽  
Author(s):  
K. L. Perry ◽  
L. Miller ◽  
L. Williams
Keyword(s):  
New York ◽  
Tomato Spotted Wilt Virus ◽  
New York State ◽  
Impatiens Necrotic Spot Virus ◽  
Spot Virus ◽  
York State ◽  
Tomato Spotted Wilt ◽  
In Vitro Plantlets ◽  
Wilt Virus

Impatiens necrotic spot virus (INSV; genus Tospovirus) was detected in experimental greenhouse-grown potatoes (Solanum tuberosum) and Nicotiana benthamiana in New York State in July and August of 2003 and 2004. Potato leaves exhibiting necrotic lesions with a concentric pattern similar to those induced by Tomato spotted wilt virus (1) were observed on cvs. Atlantic, Huckleberry, NY115, and Pentland Ivory. The presence of INSV was confirmed using double-antibody sandwich enzyme-linked immunosorbent assay and a rapid ‘ImmunoStrip’ assay (Agdia, Inc., Elkhart, IN). INSV-specific sequences were amplified from total RNA extracts using reverse transcription-polymerase chain reaction with ‘Tospovirus Group’ primers (Agdia, Inc.) and two independently amplified DNAs were sequenced. A common sequence of 355 nucleotides (GenBank Accession No. AY775324) showed 98% identity to coding sequences in an INSV L RNA. The virus was mechanically transmitted to potato and N. benthamiana and could be detected in asymptomatic, systemically infected potato leaves. Stems nodes and leaves were removed from infected potato plants, and sterile in vitro plantlets were established (2). None of the regenerated in vitro plantlets of cvs. Pentland Ivory (6 plantlets) or NY115 (5 plantlets) were infected with INSV. Two of ten regenerated cv. Atlantic plantlets initially tested positive, but INSV could not be detected after 6 months in tissue culture. In vitro tissue culture plantlets could not be established from infected cv. Huckleberry plants, even though they were consistently obtained from uninfected plants. Infected greenhouse plants were grown to maturity and the tubers harvested, stored for 6 months at 4°C, and replanted in the greenhouse. INSV could not be detected in plants from 26 cv. Huckleberry, 4 cv. NY115, or 4 cv. Atlantic tubers. Although this isolate of INSV was able to systemically infect potato, it was not efficiently maintained or transmitted to progeny tubers. This might explain why INSV has not been reported as a problem in potato production. Lastly, in both years, dying N. benthamiana provided the first sign of a widespread greenhouse infestation of INSV in a university facility housing ornamental and crop plants. INSV induced a systemic necrosis in N. benthamiana, and this host may be useful as a sensitive ‘trap’ plant indicator for natural infections in greenhouse production. References: (1) T. L. German. Tomato spotted wilt virus. Pages 72–73 in: Compendium of Potato Diseases. W. R. Stevenson et al., eds. The American Phytopathological Society, St. Paul, 2001. (2) S. A. Slack and L. A. Tufford. Meristem culture for virus elimination. Pages 117–128 in: Fundamental Methods of Plant Cell, Tissue and Organ Culture and Laboratory Operations. O. L. Gamborg and G. C. Philips, eds. Springer-Velag, Berlin, 1995.

scholarly journals First Report of Tomato spotted wilt virus on Chrysanthemum in Serbia

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 150-150 ◽  
Author(s):  
I. Stanković ◽  
A. Bulajić ◽  
A. Vučurović ◽  
D. Ristić ◽  
K. Milojević ◽  
...  
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Disease Incidence ◽  
N Gene ◽  
Impatiens Necrotic Spot Virus ◽  
Maximum Parsimony Tree ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Negative Controls ◽  
Wilt Virus

In July 2011, greenhouse-grown chrysanthemum hybrid plants (Chrysanthemum × morifolium) with symptoms resembling those associated with tospoviruses were observed in the Kupusina locality (West Bačka District, Serbia). Disease incidence was estimated at 40%. Symptomatic plants with chlorotic ring spots and line patterns were sampled and tested by double antibody sandwich (DAS)-ELISA using polyclonal antisera (Bioreba AG, Reinach, Switzerland) against the two of the most devastating tospoviruses in the greenhouse floriculture industry: Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV) (2). Commercial positive and negative controls and extracts from healthy chrysanthemum tissue were included in each ELISA. TSWV was detected serologically in 16 of 20 chrysanthemum samples and all tested samples were negative for INSV. The virus was mechanically transmitted from ELISA-positive chrysanthemum samples to five plants each of both Petunia × hybrida and Nicotiana tabacum ‘Samsun’ using chilled 0.01 M phosphate buffer (pH 7) containing 0.1% sodium sulfite. Inoculated plants produced local necrotic spots and systemic chlorotic/necrotic concentric rings, consistent with symptoms caused by TSWV (1). The presence of TSWV in ELISA-positive chrysanthemum plants and N. tabacum‘Samsun’ was further confirmed by conventional reverse transcription (RT)-PCR. Total RNAs were extracted with an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). RT-PCR was performed with the One-Step RT-PCR Kit (Qiagen) using primers TSWVCP-f/TSWVCP-r specific to the nucleocapsid protein (N) gene (4). A Serbian isolate of TSWV from tobacco (GenBank Accession No. GQ373173) and RNA extracted from a healthy chrysanthemum plant were used as positive and negative controls, respectively. An amplicon of the correct predicted size (738-bp) was obtained from each of the plants assayed, and that derived from chrysanthemum isolate 529-11 was purified (QIAqick PCR Purification Kit, Qiagen) and sequenced (JQ692106). Sequence analysis of the partial N gene, conducted with MEGA5 software, revealed the highest nucleotide identity of 99.6% (99% amino acid identity) with 12 TSWV isolates deposited in GenBank originating from different hosts from Italy (HQ830186-87, DQ431237-38, DQ398945), Montenegro (GU355939-40, GU339506, GU339508), France (FR693055-56), and the Czech Republic (AJ296599). The consensus maximum parsimony tree obtained on a 705-bp partial N gene sequence of TSWV isolates available in GenBank revealed that Serbian TSWV isolate 529-11 from chrysanthemum was clustered in the European subpopulation 2, while the Serbian isolates from tomato (GU369723) and tobacco (GQ373172-73 and GQ355467) were clustered in the European subpopulation 1 denoted previously (3). The distribution of TSWV in commercial chrysanthemum crops is wide (2). To our knowledge, this is the first report of TSWV infecting chrysanthemum in Serbia. Since chrysanthemum popularity and returns have been rising rapidly, the presence of TSWV may significantly reduce quality of crops in Serbia. References: (1) Anonymous. OEPP/EPPO Bull. 34:271, 2004. (2) Daughtrey et al. Plant Dis. 81:1220, 1997. (3) I. Stanković et al. Acta Virol. 55:337, 2011. (4) A. Vučurović et al. Eur. J. Plant Pathol. 133:935, 2012.

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