scholarly journals First Report of Tomato spotted wilt virus Infecting African Clover (Trifolium tembense) in Georgia

Plant Disease ◽  
2009 ◽  
Vol 93 (2) ◽  
pp. 202-202 ◽  
Author(s):  
N. A. Barkley ◽  
D. L. Pinnow ◽  
M. L. Wang ◽  
G. A. Pederson
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Plant Genetic Resources ◽  
Germplasm Collection ◽  
Rt Pcr ◽  
First Report ◽  
Nucleocapsid Gene ◽  
Weed Host ◽  
Tomato Spotted Wilt ◽  
Wilt Virus ◽  
Das Elisa

Tomato spotted wilt virus (TSWV; family Bunyaviridae, genus Tospovirus), which is vectored by several species of thrips (order Thysanoptera, family Thripidae), causes a destructive disease that affects many economically important host plants such as tomatoes, peppers, and peanuts. Controlling the spread of this disease is challenging, and currently, only limited strategies are available to prevent and/or control its dissemination, including early diagnosis, destruction of infected material, and elimination of the vector. TSWV has been previously reported in subterranean clover (Trifolium subterraneum), white clover (T. repens), and various unidentified wild clovers (Trifolium spp.) in North America and Australia (1,3), but never before in an African species. T. tembense (Fresen.), an herbaceous annual African clover that is mainly used for grazing, is part of the national germplasm collection housed at the Plant Genetic Resources Conservation Unit in Griffin, GA. TSWV was found naturally infecting several accessions of this species being grown for regeneration in a greenhouse during 2008. Initial putative identification of the virus was done by visual inspection of host symptoms that included ringspots, necrotic and chlorotic local lesions, sometimes mild systemic wilting, and eventually an overall decline of healthy tissue in the infected plants. This was subsequently confirmed by double-antibody sandwich (DAS)-ELISA and reverse transcription (RT)-PCR. Primers (5′-ATGTCTAAGGTTAAGCTC-3′ forward and 5′-TTAAGCAAGTTCTGTGAG-3′ reverse) targeted the nucleocapsid gene of TSWV and amplified an expected product of approximately 800 bp (2). No product was amplified in any of the negative controls. Twenty-six individuals representing twelve plant accessions (PI 517788, 517790, 517792, 517793, 517809, 517832, 517842, 517845, 517851, 517871, 517876, and 517889) were screened for TSWV. Two to three individuals were targeted from each accession. Samples were chosen on the basis of the availability of leaf tissue to perform two diagnostic assays, ELISA and RT-PCR. Samples chosen for this study were all naturally infected by thrips. All but four individuals representing two plant accessions tested positive for the virus. The RT-PCR data substantiated the DAS-ELISA results and confirmed the suspected infection. More than 26% of the positive samples naturally infected by TSWV were further characterized by purifying and sequencing (bidirectionally) the RT-PCR product on an automated CEQ 8000 sequencer (Beckman Coulter, Fullerton, CA). The resulting sequences were aligned and edited using AlignIR (LI-COR, Lincoln, NE). More than 700 bp of sequence data (GenBank Accession No. FJ183743–FJ183746) was compiled and they displayed 98% identity with deposited TSWV nucleocapsid gene sequences in GenBank, with no similarity to any other targets. To our knowledge, this is the first report of TSWV infection in T. tembense. Accessions potentially resistant to TSWV within this species were identified and need to be further substantiated. T. tembense is a wild, native clover in Africa and could serve as a weed host for infection of nearby agronomically important crops. References: (1) I. Bitterlich and L. S. MacDonald. Can. Plant Dis. Surv. 73:137, 1993. (2) R. J. Holguín-Peña and E. O. Rueda-Puente. Plant Dis. 91:1682, 2007. (3) C. R. Wilson. Plant Pathol. 47:171, 1998.

scholarly journals First Report of Tomato spotted wilt virus Infection of Tomatillo in Georgia

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1155-1155 ◽  
Author(s):  
J. C. Díaz-Pérez ◽  
H. R. Pappu
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Plant Size ◽  
Coding Region ◽  
Rt Pcr ◽  
First Report ◽  
Nucleocapsid Gene ◽  
Tomato Spotted Wilt ◽  
Pcr Product ◽  
The University ◽  
Wilt Virus

During the 2000 spring season, tomatillo (Physalis ixocarpa) plants showing chlorotic streaks on leaves were observed in an experimental plot of the University of Georgia's Coastal Plain Experiment Station in Tift County, GA. Leaf samples from 192 plants were collected. These included plants that had chlorotic streaks and those without obvious symptoms. Samples were tested by ELISA using a commercially available Tomato spotted wilt virus (TSWV) detection kit (Agdia Inc., Elkhart, IN). TSWV was found in 10 samples that had chlorotic streaks on leaves, and the remaining plants with no obvious symptoms were negative for TSWV. Infected plants were found in both cultivars, Verde Puebla and Toma Verde. The presence of the virus had no apparent effect on plant size or fruit appearance. TSWV infection of the ELISA-positive samples was further verified by immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) (1). The primer pair (5′-ATGTCTAAGGTTAAGCTC-3′ and 5′ TTAAGCAAGTTCTGTGAG-3′) represented the first and last 18 bases of the coding region of the nucleocapsid gene of TSWV, respectively, and produced approximately 800-bp PCR product (1). IC-RT-PCR gave a single DNA band of expected size and no amplification was found in the uninfected control. This is the first report of TSWV on tomatillo in Georgia. Reference: (1) R. K. Jain et al. Plant Dis. 82:900, 1998.

scholarly journals First Report of Tomato spotted wilt virus on a New Host Leuzea carthamoides in Bulgaria and the World

Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1258-1258 ◽  
Author(s):  
B. Dikova ◽  
N. Petrov ◽  
A. Djourmanski ◽  
H. Lambev
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Plant Viruses ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
The World ◽  
Systemic Symptoms ◽  
Wilt Virus ◽  
The Town ◽  
Das Elisa

The Siberian plant Leuzea carthamoides or maral root was introduced to Europe as a medicinal crop. Tomato spotted wilt virus (TSWV), genus Tospovirus, family Bunyaviridae, caused a harmful outbreak on L. carthamoides in central Bulgaria near the town of Kazanluk in 2009. In 2011, TSWV was identified on young sprouts from the rootages of L. carthamoides in the same place near the town of Kazanluk, Bulgaria, by means of indicator (test) plants, double antibody sandwich (DAS)-ELISA, and reverse transcription (RT)-PCR. Disease symptoms were small yellow spots on the young leaves grown from the tested sprouts and distortions of the leaf lamina. The old leaves had large yellow spots and necrosis, without deformations. Most of those L. carthamoides plants with such symptoms died in the second and third year. The number of the plants in the plantations decreased 20 to 40% during the 3-year period and some of these losses were from the virus disease except the environment conditions. DAS-ELISA was carried out with polyclonal TSWV antiserum of LOEWE Biochemica, GmbH, Germany. We obtained positive extinction values ODλ 405nm 0.358 ± 0.091 compared to the negative 0.053 ± 0.016 and the positive control 0.510 at a confidential interval at P ≤ 0.05. TSWV symptoms were observed on the following indicator plants according to Antignus et al. (1) and DPV/412 (2): Chenopodium quinoa, Cucumis sativus, Datura stramonium, Nicotiana glutinosa, N. rustica, N. tabacum cv. Samsun NN, and Petunia hybrida. TSWV caused on C. quinoa and on cotyledons of C. sativus cv. Delikates local chlorotic lesions only. In this TSWV differed from CMV because CMV caused systemic mosaic symptoms. Local small necrotic lesions and no systemic symptoms were observed on P. hybrida. We noticed systemic symptoms caused from TSWV on D. stramonium, N. glutinosa, N. rustica, and N. tabacum cv. Samsun NN. The systemic symptoms were chlorotic spots, concentric ring spots, and line patterns proceeding to necrosis. RT-PCR, adapted by Mumford et al. (3), was carried out on samples of L. carthamoides. Oligonucleotide primer sequences were used in accordance with Mumford et al. (3). The DNA fragment was visualized by UV trans-illumination. A fragment of the TSWV genome with a length of 276 base pairs was found in three young L. carthamoides leaf samples taken from the sprouts (marker 100 bp). The PCR fragment was sequenced and deposited to NCBI with GenBank Accession No. KC918808. PCR master mix without RNA template was used as a negative control. L. carthamoides is a newly established TSWV host in the world. To our knowledge, this is the first report of TSWV in L. carthamoides identified by RT-PCR. References: (1) Y. Antignus et al. Phytoparasitica 25:319, 1997. (2) R. Kormelink. Descriptions of Plant Viruses, p. 412, 2005. (3) R. A. Mumford et al. J. Virol. Methods 57:109, 1996.

scholarly journals First Report of Tomato spotted wilt virus in Habanero and Tabasco Peppers in Florida

Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1154-1154 ◽  
Author(s):  
M. T. Momol ◽  
H. R. Pappu ◽  
W. Dankers ◽  
J. R. Rich ◽  
S. M. Olson
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coding Region ◽  
Rt Pcr ◽  
First Report ◽  
Nucleocapsid Gene ◽  
Tomato Spotted Wilt ◽  
Habanero Pepper ◽  
Stem Lesions ◽  
Wilt Virus

In spring 2000, symptoms similar to thrips-vectored spotted wilt disease caused by Tomato spotted wilt virus (TSWV) were observed on habanero (Capsicum chinense) and tabasco (Capsicum frutescens) peppers in north Florida. Habanero peppers were from commercial fields grown for specialty markets and tabasco peppers were from research plots. Symptoms observed were leaf necrosis, fruit drop, necrotic stem lesions, and stunting. Fruit symptoms included chlorotic and necrotic spotting and distinct ring pattern and distortion. The incidence of symptomatic habanero peppers was 7 to 8% in one of the three production fields visited, and a lower incidence in two other fields (all in Jackson County). In tabasco pepper, TSWV was detected in spring and fall 1999, and spring 2000 seasons in 10 to 15% of the plants (Gadsden County). Adjacent tomato fields contained scattered plants exhibiting symptoms of TSWV. Diagnosis of TSWV from symptomatic stems, leaves, and fruit of habanero and tabasco peppers was confirmed by a double antibody sandwich enzyme linked immunosorbent assay (ELISA) using a commercially available kit (Agdia Inc., Elkhart, IN). ELISA values ranged from 1.57 to 1.95 for habanero pepper and 0.80 to 0.95 for tabasco pepper. The mean ELISA value of the negative controls was 0.001. To further verify TSWV infection, immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) was performed (1). The primer pair 5′-ATGTCTAAGGTTAAGCTC-3′ and 5′-TTAAGCAAGTTCTGTGAG-3′ represented the first and last 18 bases of the coding region of the nucleocapsid gene of TSWV, respectively, and produces approximately 800 bp PCR product (1). IC-RT-PCR gave a single DNA band of expected size in both habanero and tabasco samples, while no amplification was found in an uninfected pepper sample. This is the first report of TSWV on habanero and tabasco peppers in Florida. TSWV continues to be an economically important disease constraint to the production of tomato, pepper (C. annuum), peanut, and tobacco in the southeastern United States (observations from Georgia and Florida). Meanwhile, the known host range is expanding to include new species of cultivated vegetables. References: (1) R. K. Jain et al.. Plant Dis. 82:900, 1998.

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 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 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.

scholarly journals Tomato spotted wilt virus Identified in Desert Rose in Florida

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 526-526 ◽  
Author(s):  
S. Adkins ◽  
C. A. Baker
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
The United States ◽  
N Gene ◽  
Enzyme Linked Immunosorbent Assay ◽  
Insect Vector ◽  
Rt Pcr ◽  
Tomato Spotted Wilt ◽  
Wilt Virus ◽  
Symptomatic Plant ◽  
Das Elisa

Desert rose (Adenium obesum (Forssk.) Roem. & Schult), a member of the family Apocynaceae, is characterized by fleshy stems and leaves and colorful flowers. This exotic ornamental, originally from southeast Africa, is propagated vegetatively and is a perennial in warm climates. Virus-like foliar symptoms, including chlorotic ring and line patterns, were observed in the fall of 2004 on one of five stock plants being maintained in a greenhouse in Fort Pierce, FL. Inclusion body morphology suggested the presence of a Tospovirus in the symptomatic plant, and Tomato spotted wilt virus (TSWV) was specifically identified in this plant using a commercially available double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA; Agdia, Elkhart, IN). TSWV was not detected in symptomless desert rose plants nor was Impatiens necrotic spot virus detected in any of the plants using DAS-ELISA. Graft transmission of TSWV to other desert rose plants was successful. 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 of the symptomatic plant confirmed the diagnosis. Nucleotide and deduced amino acid sequences of a 579-bp region of the RT-PCR product were 95 to 99% and 95 to 100% identical, respectively, to TSWV N-gene sequences in GenBank. No product was amplified from symptomless plants. Since these 3-year-old plants were grown on-site from seed and only expressed symptoms 2 months following damage to the greenhouse by hurricanes Frances and Jeanne, it is likely that viruliferous thrips were introduced from local vegetable or ornamental production areas during or following the storms. To our knowledge, this is the first report of TSWV infection of desert rose in Florida, although TSWV was observed in this plant in Europe approximately 10 years ago (3,4). Because of the wide distribution of TSWV in the United States, the increasing popularity of desert rose, and the recent identification of Cucumber mosaic virus in this host (2), attention to sanitation and insect vector management is merited during desert rose propagation and production. References: (1) S. Adkins and E. N. Rosskopf. Plant Dis. 86:1310, 2002. (2) C. A. Baker et al. Plant Dis. 87:1007, 2003. (3) J. Mertelik et al. Acta Hortic. 432:368, 1996. (4) J. Th. J. Verhoeven and J. W. Roenhorst. Acta Hortic. 377:175, 1994.

scholarly journals First Report of Tomato spotted wilt virus in Soybean (Glycine max) in Georgia

Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 524-524 ◽  
Author(s):  
C. Nischwitz ◽  
S. W. Mullis ◽  
R. D. Gitaitis ◽  
A. S. Csinos
Keyword(s):  
Glycine Max ◽  
Large Scale ◽  
Tomato Spotted Wilt Virus ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
First Report ◽  
Nucleocapsid Gene ◽  
Fta Cards ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

Tomato spotted wilt virus (TSWV) is a member of the family Bunyaviridae and has a wide host range including important crops such as tomato, pepper, tobacco, peanut, and onion. In areas of Georgia, soybean (Glycine max) is double cropped between two onion crops and as a rotation crop with peanuts. Soybeans do not show any TSWV symptoms, and therefore, have not been tested on a large scale for the virus. However, because symptomless weed and crop plants provide a reservoir for TSWV and the thrips vectors (2), a survey was conducted during the summer of 2005 to evaluate the occurrence of TSWV in soybean. The survey took place in seven counties in southern Georgia with field sizes ranging between 0.4 and 20 ha (1 and 50 acres). Soybean cultivars included Haskell, DP7220, DP6770, Pioneer 97B52, and Vigoro V622NRR. Of 848 randomly selected plants tested using the double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) (Agdia, Inc., Elkhart, IN), 6.6% tested positive for TSWV. Plants testing positive ranged from seedling to the pod-setting stages. Leaves and roots of several plants tested positive, indicating a systemic infection. Soybean plants testing positive using ELISA were blotted onto FTA cards (Whatman Inc., Brentford, UK) to bind viral RNA for preservation, and the blotted samples were processed according to the manufacturer's protocol. Reverse transcription-polymerase chain reaction using punch-outs from the FTA cards and TSWV nucleocapsid gene specific forward and reverse primers (5′-TTAAGCAAGTTCTGTGAG-3′ and 5′-ATGTCTAAGGTTAAGCTC-3′), respectively (4), confirmed the identity of TSWV. TSWV has been found in soybean in other parts of the world (1) but has only been reported in the United States in a survey from Tennessee (3). To our knowledge, this is the first report of the occurrence of TSWV in soybean in Georgia. The role soybean plays as a reservoir or green bridge for thrips and TSWV is currently unknown. References: (1) A. R. Golnaraghi et al. Plant Dis. 88:1069, 2004. (2) R. L. Groves et al. Phytopathology 91:891, 2001. (3) B. S. Kennedy and B. B. Reddick. Soybean Genet. Newsl. 22:197, 1995. (4) H. R. Pappu et al. Tob. Sci. 40:74, 1996.

scholarly journals First Report of Tomato spotted wilt virus in Leek (Allium porrum) in the United States

Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 525-525 ◽  
Author(s):  
C. Nischwitz ◽  
S. W. Mullis ◽  
R. D. Gitaitis ◽  
A. S. Csinos ◽  
S. M. Olson
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
The United States ◽  
Close Relative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Allium Porrum ◽  
First Report ◽  
Nucleocapsid Gene ◽  
Fta Cards ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

Tomato spotted wilt virus (TSWV) is a member of the family Bunyaviridae. It has many important crop hosts including tomato, pepper, tobacco, peanut, and onion. In Georgia, Vidalia onions (Allium cepa), a close relative of leek, can be infected by TSWV and Iris yellow spot virus (IYSV), which is another thrips-vectored tospovirus (2). For this reason, samples of leek transplants with virus-like symptoms in one field at the border of Georgia and Florida were tested for the presence of TSWV and IYSV. The transplants had been grown from seed in a greenhouse at the same location. The sampled plants exhibited extended bleaching of leaf tips and necrotic lesions. These symptoms were also seen on onion plants infected with TSWV and IYSV. The only natural infections of leek with IYSV have been reported thus far only from Reunion Island (4) and Slovenia (1), but to our knowledge, TSWV has not been reported as a pathogen of leek. Green tissue near the necrotic lesions and bleached tips of one symptomatic leaf per plant was sampled and analyzed using a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) (Agdia, Inc., Elkhart, IN). Of 90 plants tested, eight were positive for TSWV and none were positive for IYSV. Leek samples testing positive using ELISA were blotted onto FTA cards (Whatman Inc., Brentford, UK) to bind viral RNA for preservation and then processed according to the manufacturer's protocol. Punch-outs from the FTA cards were used for reverse transcription polymerase chain reaction (RT-PCR) with the TSWV-specific forward primer (5′-TTAAGCAAGTTCTGTGAG-3′) and reverse primer (5′-ATGTCTAAGGTTAAGCTC-3′) (3) to confirm the identity of TSWV. The primers are specific to the viral nucleocapsid gene. An amplicon of the expected size (774 bp) was produced from TSWV ELISA-positive leek plants, but not from healthy controls. TSWV has been found in many plants worldwide, but to our knowledge this is the first report of TSWV infecting leek. The effect that TSWV has on leek production is currently unknown. References: (1) D. A. Benson et al. Nucleic Acids Res. 1:32 (Database issue):D23-6, 2004. (2) S. W. Mullis et al. Plant Dis. 88:1285, 2004. (3) H. R. Pappu et al. Tob. Sci. 40:74, 1996. (4) I. Robène-Soustrade et al. Online publication. New Dis. Rep. 11, 2005.

scholarly journals First Report of Tomato spotted wilt virus on Brugmansia sp. in Serbia

Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 850-850 ◽  
Author(s):  
D. Nikolić ◽  
I. Stanković ◽  
A. Vučurović ◽  
D. Ristić ◽  
K. Milojević ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Tomato Spotted Wilt Virus ◽  
N Gene ◽  
Broad Host Range ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Plant Pathol ◽  
Negative Controls ◽  
Wilt Virus

Brugmansia (Brugmansia spp.), also known as Angel's trumpet, is a perennial shrub in the Solanaceae that is a popular landscape plant in the tropics and subtropics, and potted plant in temperate regions. In April 2012, virus-like symptoms including chlorotic leaf patterns and curling followed by necrosis and distortion of leaves were observed on five outdoor-grown brugmansia plants in a private garden in Mackovac, Rasina District, Serbia. Symptomatic leaves were tested for the presence of several common ornamental viruses including Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), Cucumber mosaic virus (CMV), and Tobacco mosaic virus (TMV) by commercial double-antibody sandwich (DAS)-ELISA diagnostic kits (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls and extract from healthy brugmansia leaves were included in each ELISA. TSWV was detected serologically in all five brugmansia samples and all tested samples were negative for INSV, CMV, and TMV. The virus was mechanically transmitted from an ELISA-positive sample (41-12) to five plants of each Petuina × hybrida and Nicotiana glutinosa. Inoculated P. × hybrida plants showed local necrotic lesions and N. glutinosa showed mosaic and systemic necrosis 4 and 12 days post-inoculation, respectively, which were consistent with symptoms caused by TSWV (1). For further confirmation of TSWV infection, reverse transcription (RT)-PCR was performed with the OneStep RT-PCR (Qiagen, Hilden, Germany) using a set of TSWV-specific primers, TSWV CP-f and TSWV CP-r (4), designed to amplify a 738-bp fragment of the nucleocapsid protein (N) gene. Total RNAs from naturally infected brugmansia and symptomatic N. glutinosa plants were extracted using the RNeasy Plant Mini Kit (Qiagen). Total RNAs obtained from the Serbian tobacco isolate of TSWV (GenBank Accession No. GQ373173) and healthy brugmansia plants were used as positive and negative controls, respectively. The expected size of the RT-PCR product was amplified from symptomatic brugmansia and N. glutinosa but not from healthy tissues. The amplified product derived from the isolate 41-12 was sequenced directly after purification with the QIAquick PCR Purification kit (Qiagen), deposited in GenBank (JX468080), and subjected to sequence analysis by MEGA5 software (3). Sequence comparisons revealed that the Serbian isolate 41-12 shared the highest nucleotide identity of 99.9% (99.5% amino acid identity) with an Italian TSWV isolate P105/2006RB (DQ915946) originating from pepper. To our knowledge, this is the first report of TSWV on brugmansia in Serbia. Due to the increasing popularity and economic importance of brugmansia as an ornamental crop, thorough inspections and subsequent testing for TSWV and other viruses are needed. This high-value ornamental plant may act also as reservoir for the virus that can infect other ornamentals and cultivated crops, considering that TSWV has a very broad host range (2). References: (1) Anonymous. OEPP/EPPO Bull. 34:271, 2004. (2) G. Parrella et al. J. Plant Pathol. 85:227, 2003. (3) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011. (4) A. Vučurović et al. Eur. J. Plant Pathol. 133:935, 2012.

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