The First Report of Tomato spotted wilt virus on Gerbera and Chrysanthemun in Venezuela

Gerbera (Gerbera jamesonii) and Chrysanthemum (family Asteraceae) are two of the top 10 cut flowers of the world, with great commercial value. Since 1998, Venezuela began a floral industry to produce and export fresh cut gerbera and chrysanthemum, with 40% of nurseries concentrated in Altos Mirandinos (Miranda State, north central region of the country). For the past 2 years, greenhouse-grown gerbera and chrysanthemum have been observed displaying symptoms resembling those associated with tospoviruses. Symptomatic plants showed concentric rings, irregular chlorotic blotches, and deformation on leaves. Disease incidence was estimated at 30%. Mechanical inoculation with extracts of symptomatic leaves reproduced the typical concentric ring symptoms on indicator plants Arachis hypogaea L. cv. San Martín, Capsicum chinense, and G. jamesonii 6 to 15 days after inoculation. In initial tests, leaves from each 30 symptomatic gerbera and chrysanthemum species from several greenhouse facilities in Altos Mirandinos reacted positively when tested by DAS-ELISA with polyclonal antisera (ATCC, Rockville, MD) raised against Tomato spotted wilt virus (TSWV). Total RNA was extracted with the RNeasy Plant Mini kit (QIAGEN, Hilden, Germany) from two gerbera and two chrysanthemum ELISA-positive samples. The TSWV coat protein gene was amplified by conventional reverse transcription (RT)-PCR using primers CP1 TSWV (TTAACTTACAGCTGCTTT) and CP2 TSWV (CAAAGCATATAAGAACTT) (1). A single DNA product of ~823 bp was amplified from all samples. RT-PCR products were directly sequenced in both orientations and sequences were deposited in GenBank (Accession Nos. KF146700 and KF146701 derived from chrysanthemum, KF146702 and KF146703 derived from gerbera). The resulting sequences showed over 99% identity with each other. and were found to be closely related (over 99%) with TSWV isolates deposited in GenBank originating from different hosts from France (FR693058, FR693055), Montenegro (GU339506, GU339508, GU355940), Italy (HQ830187), New Zealand (KC494501), South Korea (KC261967), and the United States (AY744476). To our knowledge, this is the first confirmed report of TSWV infecting gerbera and chrysanthemum in Venezuela. The relatively widespread occurrence of TSWV in Miranda State underscores the need for systematic surveys to assess its incidence and impact on ornamental crops so that appropriate management tactics can be developed. Reference: (1) R. A. Mumford et al. J. Virol. Methods 57:109, 1996.

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Evaluation of Arachis Species for Resistance to Tomato Spotted Wilt Virus

Peanut Science ◽

10.3146/pnut.29.2.0001 ◽

2002 ◽

Vol 29 (2) ◽

pp. 79-84 ◽

Cited By ~ 14

Author(s):

J. H. Lyerly ◽

H. T. Stalker ◽

J. W. Moyer ◽

K. Hoffman

Keyword(s):

Tomato Spotted Wilt Virus ◽

Insect Pests ◽

Disease Incidence ◽

Diploid Species ◽

Breeding Programs ◽

Arachis Species ◽

Tomato Spotted Wilt ◽

Arachis Hypogaea L ◽

Cultivated Species ◽

Wilt Virus

Abstract Tomato spotted wilt virus (TSWV) is an important plant pathogen with a wide host range, including the domesticated peanut (Arachis hypogaea L.). After initial outbreaks on peanut during the 1980s, the virus has spread to all peanut-producing states in the U.S. TSWV is transmitted by several species of thrips which are difficult to control with insecticides; therefore, control of TSWV most likely will come from selecting resistant genotypes in breeding programs. Although moderate levels of resistance have been discovered in A. hypogaea, complete virus resistance has not been found. Several Arachis species have desirable genes for plant resistances and tolerate many disease and insect pests better than the cultivated species. The objectives of this study were to (a) evaluate TSWV disease incidence and severity in accessions of Arachis species, and (b) compare levels of TSWV resistance in diploid species to selected A. hypogaea genotypes. In this study, 46 diploid Arachis spp. accessions were evaluated in the greenhouse by artificial inoculation tests for resistance to TSWV. Nine Arachis accessions were observed with no disease symptoms when TSWV isolate 10 was used as opposed to A. hypogaea lines that ranged from moderately to highly susceptible. Additional testing with more virulent isolates identified A. diogoi accession GKP 10602 and A. correntina accession GKP 9530 as highly resistant to the virus. These two accessions are being used as parents in crossing programs to incorporate TSWV resistance genes into A. hypogaea.

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First Report of Tomato spotted wilt virus on Chrysanthemum in Serbia

Plant Disease ◽

10.1094/pdis-08-12-0778-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 150-150 ◽

Cited By ~ 7

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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Tomato spotted wilt virus Identified in Desert Rose in Florida

Plant Disease ◽

10.1094/pd-89-0526c ◽

2005 ◽

Vol 89 (5) ◽

pp. 526-526 ◽

Cited By ~ 5

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.

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Peanut plants in Queensland infected with tomato spotted wilt virus

Australian Journal of Agricultural Research ◽

10.1071/ar9610239 ◽

1961 ◽

Vol 12 (2) ◽

pp. 239 ◽

Cited By ~ 12

Author(s):

K Helms ◽

NE Grylls ◽

GS Purss

Keyword(s):

Arachis Hypogaea ◽

Tomato Spotted Wilt Virus ◽

Disease Incidence ◽

Infective Virus ◽

Tomato Spotted Wilt ◽

Arachis Hypogaea L ◽

Tagetes Minuta ◽

New Hosts ◽

The Mean ◽

Wilt Virus

A disease of peanut (Arachis hypogaea L.) in Queensland, previously referred to as "chlorosis", was identified as being caused by the tomato spotted wilt virus. Symptoms of the disease on peanut are described. Erigeron bonariensis L., Tagetes minuta L., and Trifolium subterraneurn L. are recorded as new hosts. The disease was not seed-transmitted, although seeds developed lesions and were malformed. Infective virus was found in tissues of the integuments of immature seed. The maximum disease incidence recorded in an individual crop was 5.5%. The mean seed yield of diseased plants was 12.7 g per plant, whereas that of healthy plants was 128.2 g.

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Transgressive Segregation and Long-Term Consistency for High TSWV Field Resistance in the ‘Georgia-06G’ Peanut Cultivar

Plant Health Progress ◽

10.1094/php-04-18-0018-rs ◽

2018 ◽

Vol 19 (3) ◽

pp. 201-206 ◽

Cited By ~ 1

Author(s):

W. D. Branch ◽

A. K. Culbreath

Keyword(s):

Disease Incidence ◽

The United States ◽

Transgressive Segregation ◽

Field Resistance ◽

Breeding Lines ◽

Tomato Spotted Wilt ◽

Arachis Hypogaea L ◽

Production Area ◽

Wilt Virus

Tomato spotted wilt disease caused by tomato spotted wilt virus (TSWV) is a major peanut (Arachis hypogaea L.) production problem in the United States. TSWV has become endemic since the mid-1980s in the southeastern U.S. peanut production area. ‘Georgia-06G’ is a large-seeded, TSWV-resistant, runner-type peanut cultivar, whereas ‘Georgia Greener’ is a sister line with a smaller seed size than Georgia-06G. Both Georgia-06G and Georgia Greener have greater TSWV general field resistance with higher yields, total sound mature kernels grade, and dollar values than the parents (‘Georgia Green’ and ‘C-99R’), which shows transgressive segregation for these desirable quantitative traits. Therefore, the objective of this 12-year study was to evaluate the higher TSWV host-plant resistance found in the Georgia-06G over time compared with many other runner- and virginia-type peanut cultivars and advanced breeding lines. Despite yearly variability in TSWV and total disease incidence, the data indicate that Georgia-06G exhibited long-term high TSWV and total disease field resistance at midseason and late season, respectively. Georgia-06G was also found to have among the highest pod yield and dollar value every year. The higher general TSWV field resistance of Georgia-06G appears to be consistent across many years and locations, similar to the moderate TSWV resistance in Georgia Green.

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Genotype × Seeding Rate Interaction among TSWV-Resistant, Runner-Type Peanut Cultivars1

Peanut Science ◽

10.3146/pnut.30.2.0009 ◽

2003 ◽

Vol 30 (2) ◽

pp. 108-111 ◽

Cited By ~ 19

Author(s):

W. D. Branch ◽

J. A. Baldwin ◽

A. K. Culbreath

Keyword(s):

Coastal Plain ◽

Tomato Spotted Wilt Virus ◽

Disease Incidence ◽

Seeding Rate ◽

Green It ◽

Tomato Spotted Wilt ◽

Arachis Hypogaea L ◽

Peanut Production ◽

Rate Interaction ◽

Wilt Virus

Abstract Tomato spotted wilt virus (TSWV) resistant, runner-type peanut (Arachis hypogaea L.) cultivars are the most important defense to control spotted wilt disease in southeast U.S. peanut production. The objective of this 3-yr (1999–01) study was to evaluate six TSWV-resistant, runner-type cultivars (Southern Runner, Florida MDR 98, C-99R, ViruGard, Georgia Green, and Georgia-OIR) at three different seeding rates (3, 5, and 7 seed/30.5 cm) in single conventional row patterns for possible genotype (GE) × seeding rate (SR) interaction at the Univ. of Georgia, Coastal Plain Experiment Station. The combined split-plot analyses of variance resulted in highly significant (P ≤ 0.01) GE × SR interaction, which indicates that not all six runner-type cultivars performed the same at each of these three seeding rates. A good example was the TSWV-resistant, runner-type peanut cultivar Georgia Green. It performed subpar at the below normal or lowest seeding rate; whereas at the highest seeding rate, Georgia Green and Georgia-OIR produced the highest pod yields and dollar value returns per hectare among all of these runner-type cultivars. TSWV disease incidence was also significantly lower for the TSWV-resistant Georgia Green cultivar at each of the two higher seeding rates compared to the lowest seeding rate.

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First Report of Tomato spotted wilt virus Infecting Onion and Garlic in Serbia

Plant Disease ◽

10.1094/pdis-02-12-0157-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 918-918 ◽

Cited By ~ 5

Author(s):

I. Stanković ◽

A. Bulajić ◽

A. Vučurović ◽

D. Ristić ◽

K. Milojević ◽

...

Keyword(s):

Tomato Spotted Wilt Virus ◽

Disease Incidence ◽

Sandwich Elisa ◽

Thrips Tabaci ◽

N Gene ◽

Pcr Analysis ◽

Rt Pcr ◽

Tomato Spotted Wilt ◽

Negative Controls ◽

Wilt Virus

In June 2011, extensive bleaching and numerous small whitish spots on leaves were observed in an onion (Allium cepa) seed crop as well as chlorotic spots and streaks in the neighboring garlic (A. sativum) bulb crop in the Aleksandrovo locality (Central Banat District, Serbia). Affected plants occurred throughout the field and disease incidence was estimated at 60% in the onion and 40% in the garlic crop. A high population of Thrips tabaci that was found in both crops, and local necrotic spots on Petunia × hybrida mechanically inoculated with infected onion or garlic sap by a chilled 0.01 M phosphate buffer, pH 7.0, containing 0.1% sodium sulfite (1), suggested the presence of a Tospovirus. For these reasons, sampled symptomatic onion and garlic plants were tested for the presence of Tomato spotted wilt virus (TSWV) and Iris yellow spot virus (IYSV) using commercial double-antibody sandwich-ELISA diagnostic kits (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls and extracts from healthy onion and garlic tissue were included in each ELISA. Of the 18 onion and 10 garlic plants tested, 16 and 7 samples, respectively, were positive for TSWV, and all were negative for IYSV. The identity of TSWV was further confirmed by conventional reverse transcription (RT)-PCR analysis. Total RNAs were extracted with an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was performed with the One-Step RT-PCR Kit (Qiagen) using TSWV-specific forward (5′-GGTTAAGCTCACTAAGAAARCA-3′) and reverse primers (5′-TTTAACYCCRAACATTTCATAGA-3′), designed to amplify a 738-bp fragment of the nucleocapsid protein (N) gene. Total RNAs obtained from plants infected with a Serbian isolate of TSWV (GenBank Accession No. GQ373173) and healthy onion garlic plants were used as positive and negative controls, respectively. An amplicon of the expected size was produced from the 16 onion and 7 garlic ELISA-positive plants, but not from healthy controls. The amplified products derived from the two selected isolates, 114-11 from onion and 115-11 from garlic, were sequenced directly after purification with the QIAquick PCR Purification kit (Qiagen); the sequences obtained were allocated GenBank Accession Nos. JQ619234 and JQ619235, respectively. Sequence analysis of the partial N gene, conducted with MEGA5 software (4), revealed 99.9% nucleotide identity (100% amino acid identity) between the two Serbian Allium isolates. Serbian onion and garlic isolates showed the highest nucleotide identities of 100% and 99.9% with Serbian summer squash isolate (JF303081) and tobacco isolate from Montenegro (GU369729), respectively. Well-established in many European countries, TSWV has been reported as an important constraint to the production of tomato, pepper, tobacco, and ornamentals (2), but the information on TSWV naturally infecting Allium spp. is limited. The presence of TSWV on onion and garlic in Serbia revealed that its known host range has expanded in Europe. To our knowledge, other than Marchoux's unpublished data (3), there are no other reports of garlic as a natural host of TSWV. The TSWV presence on Allium spp. represents a serious threat for these crops in Serbia, considering that it is prevalent in other crops in the area and its vectors are widespread. References: (1) Anonymous. OEPP/EPPO Bull. 34:271, 2004. (2) H. R. Pappu et al. Virus Res. 141:219, 2009. (3) G. Parrella et al. J. Plant Pathol. 85:227, 2003. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

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

Fitopatologia Brasileira ◽

10.1590/s0100-41582002000300008 ◽

2002 ◽

Vol 27 (3) ◽

pp. 285-291 ◽

Cited By ~ 9

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).

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

Peanut Science ◽

10.3146/ps08-024.1 ◽

2009 ◽

Vol 36 (2) ◽

pp. 133-137 ◽

Cited By ~ 1

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.

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