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 Together with a previous observation of TSWV in north Florida nursery stock (L. Breman, unpublished), this represents, to our knowledge, the first report of TSWV infection of blackberry lily in North America although TSWV was observed in plants of this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 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.

scholarly journals Outbreak of Tomato spotted wilt virus in Tomato in Kenya

Plant Disease ◽  
2001 ◽  
Vol 85 (10) ◽  
pp. 1123-1123 ◽  
Author(s):  
A. W. Wangai ◽  
B. Mandal ◽  
H. R. Pappu ◽  
S. Kilonzo
Keyword(s):  
Host Range ◽  
Tomato Spotted Wilt Virus ◽  
Virus Disease ◽  
N Gene ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Tomato Spotted Wilt ◽  
Wide Host Range ◽  
Test Kit ◽  
Wilt Virus

Tomato spotted wilt virus (TSWV) of the genus Tospovirus, family Bunyaviridae (1), causes an economically important virus disease in tomato in several parts of the world. The virus has a wide host range that includes numerous crops and weeds and is transmitted by at least seven species of thrips. Tomato crops in the Subukia, Bahati, and Kabazi areas of the Nakuru District in Kenya were affected by a disease suggestive of TSWV infection during the November 1999 to March 2000 tomato-growing season. Farmers reported up to 80% losses of their potential yields. Characteristic symptoms were noticed on fruits, especially when they were green. Distinct concentric rings on fruits, which later turned into brown, uneven ripening, were the most visible symptoms. Foliage did not develop pronounced symptoms, but mild bronzing was observed in a few cultivars. However, foliage senesced prematurely, starting with older leaves. Foliar symptoms were mistaken for blight infection, and as a result, excessive fungicides were applied that failed to manage the disease. To test for TSWV infection, tomato leaf samples collected from the fields were tested initially with a TSWV test kit (HortiTech, Horticulture Research International, Wellesbourne, UK), and the results were confirmed by double-antibody sandwich-enzyme-linked immunosorbent assay with antibodies from Agdia Inc. (Elkhart, IN). Further molecular characterization was done using reverse transcription-polymerase chain reaction (RT-PCR). Total RNA was extracted from symptomatic leaves of tomato cv. Money Maker using the RNeasy mini kit (Qiagen Inc., Valencia, CA). Using primers 5′ TTAAGC AAGTTCTGTGAG 3′ and 5′ ATGTCTAAGGTTAAGCTC 3′ specific to the nucleoprotein (N) gene of TSWV, the N gene was amplified by RT-PCR (2). A 777-bp product of the expected size was obtained from symptomatic plants, whereas no amplification was obtained from noninfected tomato. The PCR product was cloned into pGEM-T Easy (Promega, Madison, WI) and sequenced. A search of GenBank revealed a sequence identity of 95 to 99% with the N genes of known TSWV isolates. To our knowledge, this is the first report TSWV infection of tomato in Kenya. Considering its wide host range, future surveys should be directed toward estimating its incidence in tomato and other TSWV-susceptible crops, such as Irish potatoes, pepper, peanut (groundnut), beans, and a wide variety of ornamental cut flowers in Kenya. References: (1) J. W. Moyer. Tospoviruses (Bunyaviridae). Pages 1803–1807 in: Encyclopedia of Virology. A. Granoff and R. G. Webster, eds. Academic Press, San Diego, CA, 1999. (2) Jain et al. Plant Dis. 82:900, 1998.

scholarly journals First Report of Tomato spotted wilt virus on Pepper in Montenegro

Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 882-882 ◽  
Author(s):  
J. Zindović ◽  
A. Bulajić ◽  
B. Krstić ◽  
M. Ciuffo ◽  
P. Margaria ◽  
...  
Keyword(s):  
Amino Acid ◽  
Tomato Spotted Wilt Virus ◽  
Nucleotide Identity ◽  
N Gene ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Italian Isolate ◽  
Negative Controls ◽  
Wilt Virus

In April 2009, chlorotic and necrotic ring spots, chlorotic line patterns, and stunting were observed on greenhouse-grown pepper plants in the vicinity of Podgorica, Montenegro. Disease symptom incidence was estimated at 40%. Symptomatic leaves were tested for the presence of Tomato spotted wilt virus (TSWV) with a commercial double-antibody sandwich (DAS)-ELISA diagnostic kit (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls were included in each ELISA. TSWV was detected serologically in 33 of 75 pepper samples. The virus was mechanically transmitted from ELISA-positive pepper samples to Nicotiana tabacum cv. Samsun using chilled 0.05 M phosphate buffer (pH 7) containing 0.1% sodium sulfite (1). Inoculated test plants produced chlorotic and necrotic concentric rings and necrotic spots, consistent with symptoms caused by TSWV on N. tabacum. For further confirmation of TSWV infection, reverse transcription (RT)-PCR was performed with the One-Step RT-PCR Kit (Qiagen, Hilden, Germany) using three sets of primers: S70-for/S890-rev (2) and S574-for/S1433-rev (3), both specific to the nonstructural (NSs) gene; and S1983-for/S2767-rev (2), specific to the nucleocapsid protein (N) gene. Total RNAs from naturally infected pepper and symptomatic N. tabacum cv. Samsun plants were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Total RNAs obtained from the Italian isolate of TSWV (GenBank Accession No. DQ398945) and healthy tobacco plants were used as positive and negative controls, respectively. The expected sizes of the RT-PCR products (820, 859, and 784 bp) were amplified from symptomatic pepper samples but not from healthy tissues. The PCR product obtained from isolate Is-344 using primers specific to N gene was purified by a QIAquick PCR Purification Kit (Qiagen), cloned into the pGEM-T Easy Vector (Promega, Madison, WI) and sequenced in both directions using the same primer pair as in RT-PCR. The sequences amplified with the two primer pairs specific to the NSs gene were obtained by direct sequencing (Bio-Fab Research Srl, Pomezia, Italy) and joined using MEGA4 software. Sequence analysis of the complete N gene (777 bp; GenBank Accession No. GU369717) revealed that the TSWV isolate originating from Montenegro shared 98.2 to 99.7% nucleotide identity (98.1 to 100% amino acid identities) with corresponding TSWV sequences deposited in GenBank. The Montenegrin isolate Is-344 was most closely related to Italian isolates from tomato (GU369725) and eggplant (GU369720). The partial (1,257 bp) nucleotide sequence of NSs gene (GU369737) showed 96 to 99.8% nucleotide identity (96.9 to 100% amino acid identity) with previously reported TSWV sequences, and in this case the highest identity was with French isolates from tomato (FR692835) and lettuce (FR692831). To our knowledge, this is the first report on the occurrence of TSWV in Montenegro. Data of this study sheds light on the importance of further survey studies and inspections of TSWV-susceptible crops cultivated in Montenegro. References: (1) Anonymous. OEPP/EPPO Bull. 29:465, 1999. (2) W. P. Qiu et al. Virology 244:186, 1998. (3) M. Tsompana et al. Mol. Ecol. 14:53, 2005.

scholarly journals First Report of Tomato spotted wilt virus on Gloxinia in Bosnia and Herzegovina

Plant Disease ◽  
2013 ◽  
Vol 97 (3) ◽  
pp. 429-429 ◽  
Author(s):  
V. Trkulja ◽  
J. Mihić Salapura ◽  
B. Ćurković ◽  
I. Stanković ◽  
A. Bulajić ◽  
...  
Keyword(s):  
Bosnia And Herzegovina ◽  
Tomato Spotted Wilt Virus ◽  
Amino Acid Identity ◽  
N Gene ◽  
Impatiens Necrotic Spot Virus ◽  
Post Inoculation ◽  
Rt Pcr ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Wilt Virus

In June and July 2012, symptoms resembling those caused by a tospovirus infection were observed on the greenhouse-grown gloxinia (Sinningia speciosa Benth. and Hook.) in the Lijevče polje, in the vicinity of Banja Luka (Bosnia and Herzegovina). Infected plants exhibited chlorotic ring spots and chlorotic and necrotic patterns followed by necrosis and distortion of leaves. Disease symptom incidence was estimated at 30% out of 400 inspected plants. Symptomatic leaves were collected and tested by double-antibody sandwich (DAS)-ELISA test using commercial polyclonal antisera (Bioreba AG, Reinach, Switzerland) for two of the most important tospoviruses in the greenhouse production of ornamentals: Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV) (2). TSWV was detected serologically in 27 out of 30 tested gloxinia samples, and all were negative for INSV. Symptomatic leaves of five selected ELISA-positive gloxinia plants were separately ground in chilled 0.01 M phosphate buffer (pH 7) containing 0.1% w/v sodium sulphite and were mechanically inoculated on five plants of Petunia × hybrida. All inoculated plants produced typical symptoms of TSWV (1), necrotic spots on inoculated leaves in 2 to 5 days post-inoculation. For further confirmation of TSWV infection, total RNAs were extracted using the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) from all 27 infected gloxinia plants and tested by reverse transcription (RT)-PCR assay. A 738-bp fragment of TSWV nucleocapsid (N) gene was amplified with One-Step RT-PCR Kit (Qiagen) using primer pairs TSWV CP-f and TSWV CP-r (4). Total RNAs from Serbian tobacco TSWV isolate (GenBank Accession No. GQ373173) and RNA extract from healthy gloxinia plants were used as positive and negative controls, respectively. Amplicons of the expected size were obtained from all 27 naturally infected gloxinia plants, while no amplification products were obtained from the healthy control. After the purification with QIAquick PCR Purification Kit (Qiagen), the RT-PCR product obtained from one selected isolate 160-12 was sequenced directly in both directions and submitted to GenBank (JX468079). Sequence analysis of the partial N gene, conducted by MEGA5 software (3), from isolate 160-12 showed the highest nucleotide identity of 99.7% (100% amino acid identity) with eight pepper isolates of TSWV from Spain (FR693229, FR693231, FR693152-153, FR693078, FR693081, FR693089, and FR693092). To our knowledge, this is the first report on the occurrence of TSWV in Bosnia and Herzegovina. The presence of this harmful pathogen into a new area could have a serious threat to intensive and increasing production of ornamentals and numerous other TSWV susceptible species in Bosnia and Herzegovina. The discovery of TSWV on gloxinia should prompt more surveys, thorough inspections, and subsequent testing of other TSWV susceptible plants cultivated in Bosnia and Herzegovina. References: (1) Anonymous. OEPP/EPPO Bull. 34:271, 2004. (2) Daughtrey et al. Plant Dis. 81:1220, 1997. (3) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011. (4) A. Vučurović et al. Eur. J. Plant Pathol. 133:935, 2012.

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.

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.

scholarly journals First Report of Tomato spotted wilt virus on Soybean in Iran

Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1290-1290 ◽  
Author(s):  
A. R. Golnaraghi ◽  
N. Shahraeen ◽  
R. Pourrahim ◽  
Sh. Ghorbani ◽  
Sh. Farzadfar
Keyword(s):  
Glycine Max ◽  
Tomato Spotted Wilt Virus ◽  
Chenopodium Quinoa ◽  
Datura Stramonium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mechanical Inoculation ◽  
First Report ◽  
Tomato Spotted Wilt ◽  
Soybean Leaves ◽  
Wilt Virus

During the summers of 1999 and 2000, 3,110 soybean (Glycine max) leaf samples were randomly collected from soybean fields in the Ardebil, Goletan, Khuzestan, Lorestan, and Mazandaran provinces of Iran. Tomato spotted wilt virus (TSWV) was detected in leaf samples by TSWV-specific polyclonal antibody (As-0526 and As-0580, DSMZ, Braunschweig, Germany) in double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Mechanical inoculation of 26 plant species (10 plants per species) and cultivars with extracts of positive leaf samples produced necrotic local lesions in Beta vulgaris, Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Phaseolus vulgaris cv. Talash, Vicia faba, and Vigna unguiculata cv. Mashad; produced systemic chlorosis followed by necrosis in Datura stramonium, D. metel, Nicotiana rustica, N. tabacum cv. Samsun, N. glutinosa, N. bentamiana, and Glycine max cv. Hill; and produced chlorosis, stunting, and bud necrosis in Arachis hypogaea (peanut). Plants developing these symptoms following mechanical inoculation with extracts from original soybean leaves were positive in ELISA for TSWV. ELISA results indicate that the overall incidence of TSWV on soybean in the five provinces was 5.4%. TSWV has been reported in potato (2) and tomato (1) from Iran, but to our knowledge, this is the first report of the occurrence of TSWV on soybean in Iran. References: (1) K. Bananej et al. Iran. J. Plant Pathol. 34:30, 1998. (2) R. Pourrahim et al. Plant Dis. 85:442, 2001.

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.

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