scholarly journals First Report of Tomato spotted wilt virus and Impatiens necrotic spot virus in Slovenia

Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1288-1288 ◽  
Author(s):  
I. Mavrič ◽  
M. Ravnikar

In July 2000, concentric necrotic rings and patterns were observed on greenhouse-grown pepper (Capsicum anuum L. ‘Blondi’). Symptoms were present only on lower leaves, not on young leaves or fruits. Typical tospovirus particles using electron microscopy were observed in leaf-dip preparations of symptomatic leaves. Impatiens necrotic spot virus (INSV) was detected in symptomatic but not in asymptomatic tissues using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with polyclonal antiserum (Loewe Biochemica, Sauerlach, Germany). Nicotiana benthamiana, N. rustica, and Petunia sp. were mechanically inoculated with sap of symptomatic leaves. Local and systemic symptoms were observed only on N. benthamiana. Tomato spotted wilt virus (TSWV) infections were later confirmed in some pepper and tomato plants with distinct systemic symptoms and in greenhouse-grown chrysanthemums, calla lilies, cyclamen and spatiphylum using DAS-ELISA with polyclonal antiserum. Severe systemic symptoms were observed only on some chrysanthemum cultivars, where the infection rate of TSWV was between 80 and 100%. Such TSWV-infected plants were not marketable. Symptomless infections by the same virus were also found. At one location, calla lilies were heavily infected by TSWV, but only local symptoms on leaves were observed. INSV was also confirmed using DAS-ELISA on different chrysanthemum cultivars. Mixed infections of TSWV and INSV were detected using DAS-ELISA on calla lilies with local necrotic rings and patterns on leaves and on Impatiens walerana with systemic necrosis. A limited number of weeds from the vicinity of greenhouses containing symptomatic plants were tested for the presence of TSWV using DAS-ELISA, and only Artemisia vulgaris was infected. Both viruses, TSWV from chrysanthemum and INSV from pepper, were isolated on test plants, and their identity was confirmed using DAS-ELISA. For further verification of TSWV and INSV infection, immunocapture reverse-transcription polymerase chain reaction was performed using general tospovirus primers (1). Amplification products of the expected size were detected and sequenced. Comparison of nucleic acid sequences of amplification products with viral sequence databases confirmed the identities of both viruses. To our knowledge, this is the first report of TSWV and INSV infection in ornamental and vegetable plants in Slovenia. Reference: (1) R. J. Weekes et al. Acta Hortic. 431:159, 1996.

2003 ◽  
Vol 4 (1) ◽  
pp. 40 ◽  
Author(s):  
N. Martínez-Ochoa ◽  
A. S. Csinos ◽  
E. B. Whitty ◽  
A. W. Johnson ◽  
M. J. Parrish

Tomato spotted wilt caused by Tomato spotted wilt virus (TSWV) continues to be a serious disease problem on tobacco (Nicotiana tabacum L.), peanut (Arachis hypogaea L.), tomato (Lycopersicon esculentum Mill.), and pepper (Capsicum annum L.) in the southeastern United States. Impatiens necrotic spot virus (INSV, formerly known as TSWV-I) is an emerging virus found mostly in greenhouse production of ornamentals and is also vectored by thrips. A few years ago INSV was detected in peanut in Georgia and Texas and its occurrence appears to be increasing). Mixed infections of TSWV and INSV in tobacco have been observed within the last two years in North Carolina and Kentucky. Our objective was to sample several locations in Georgia, Florida, South Carolina and Virginia to confirm and report the presence of natural TSWV and INSV mixed infections in tobacco. Accepted for publication 14 March 2003. Published 17 April 2003.


Author(s):  
Fatma Şafak ◽  
Muharrem Arap Kamberoğlu

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.


Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1258-1258 ◽  
Author(s):  
B. Dikova ◽  
N. Petrov ◽  
A. Djourmanski ◽  
H. Lambev

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.


Plant Disease ◽  
2007 ◽  
Vol 91 (11) ◽  
pp. 1515-1515 ◽  
Author(s):  
C. A. Baker ◽  
D. Davison ◽  
L. Jones

In October 2006 (Arcadia, FL) and January 2007 (Sorrento, FL), several white Phalaenopsis orchids with large chlorotic/necrotic ringspot symptoms were sent to the Division of Plant Industry, Gainesville, FL. Symptomatic leaf tissues were tested with the Agdia immunostick-comb (Agdia, Elkhart, IN) for Impatiens necrotic spot virus (INSV), Tomato spotted wilt virus (TSWV), Cucumber mosaic, and Tobacco mosaic virus. Plants from the nursery in Sorrento, FL tested positive for TSWV, while those from the nursery in Arcadia, FL tested positive for INSV. Symptomless leaves from the infected plants tested negative for the viruses with the immunostick-comb. The plants also were tested for TSWV and INSV by double-antibody (DAS)-ELISA (Agdia Inc.) with the same results. Total RNA was extracted from one symptomatic orchid leaf from each nursery. Reverse transcription (RT)-PCR was performed with the universal tospovirus primer set BR60and BR65 (1). PCR bands of the expected size were amplified from each leaf. PCR products were sequenced directly. The orchid leaf that tested positive for TSWV by ELISA produced a 495-bp sequence with 97% identity to several isolates of the TSWV nucleocapsid protein gene listed in GenBank (Accession Nos. AY744479, AY8770391, DQ376185, and AF02659). The orchid leaf that tested positive for INSV by ELISA produced a 396-bp sequence with 98 to 99% identity to several isolates of the INSV nucleocapsid protein gene (Accession Nos. D00914, DQ425096, X66972, and AD109100). Although these viruses have been reported a few times in orchids previously (2,3), to our knowledge, this is the first time they have been reported in this host in Florida. In addition, white Phalaenopsis spp. appears to be a local lesion host and not a systemic host for these viruses. References: (1) M. Eiras et al. Fitopatol. Bras. 26:170, 2001. (2) J. S. Hu et al. Plant Dis. 77:464, 1993. (3) S. T. Koike and D. E. Mayhew. Orchids 70:746, 2001.


Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 771-771 ◽  
Author(s):  
R. Pourrahim ◽  
A. R. Golnaraghi ◽  
Sh. Farzadfar

There have been an increasing number of records of the natural infection of various crops and ornamentals in Iran with Impatiens necrotic spot virus (INSV), Tomato spotted wilt virus (TSWV), and Tomato yellow fruit ring virus (TYFRV), a recently proposed species in the genus Topovirus (3). TYFRV, originally believed to be TSWV and named as such, has been previously reported to occur in Iranian potato fields (2). During the growing seasons of 2004 to 2006, surveys were conducted in potato fields in different potato-producing (Solanum tuberosum) provinces of Iran (Ardabil, Azarbayejan-e-sharqi, Chaharmahal-va-bakhtiyari, Esfahan, Hamedan, Kerman, Khorasan, Khuzestan, Lorestan, Tehran, Qazvin, and Zanjan) to detect the presence of Tospovirus spp. infecting this crop, including Groundnut ringspot virus (GRSV), INSV, Iris yellow spot virus (IYSV), Tomato chlorotic spot virus (TCSV), TSWV, TYFRV, and Watermelon silver mottle virus (WSMoV). Overall, 186 fields were surveyed, and 2,823 potato leaf samples from plants showing tospovirus-like symptoms of chlorotic or necrotic spots, chlorosis, and necrosis were collected before or through the flowering stage, approximately 50 to 90 days after planting. Each leaf sample was tested by double-antibody sandwich (DAS)-ELISA using specific antisera (Bioreba, Reinach, Switzerland; Loewe, Sauerlach, Germany; DSMZ, Braunschweig, Germany) for the presence of the aforementioned tospoviruses. TYFRV, TSWV, and INSV were found in 24.0, 4.1, and 0.4% of the samples collected from 133, 51, and 7 fields surveyed, respectively. None of the samples had a positive reaction in ELISA to GRSV, IYSV, TCSV, and WSMoV. To confirm this testing, a number of the leaf samples that were found to be positive for INSV, TSWV, and TYFRV in ELISA tests were mechanically inoculated on Petunia × hybrid and Nicotiana benthamiana; the inoculated plants showed typical necrotic local lesions of tospoviruses and chlorotic or necrotic spots followed by systemic infection, respectively; their infection was subsequently confirmed by ELISA. The samples also were tested by reverse transcription-PCR technique using previously described specific primers (1,4). The PCR reaction resulted in the specific amplification of a 0.59-, 0.71-, and 0.67-kb (or 1.2-kb) fragment of INSV, TSWV, and TYFRV RNAs, respectively. This study showed that tospoviruses, especially TYFRV, are widespread in Iranian potato fields. It is hoped that the results may help us to improve a seed potato certification program in the future. To our knowledge, this is the first report of INSV and TSWV from potatoes in Iran. References: (1) A. R. Golnaraghi et al. Plant Dis. 92:1280, 2008. (2) R. Pourrahim et al. Plant Dis. 84:442, 2001. (3) S. Winter et al. Plant Pathol. 55:287, 2006. (4) H. Uga and S. Tsuda. Phytopathology 95:166, 2005.


2002 ◽  
Vol 27 (3) ◽  
pp. 285-291 ◽  
Author(s):  
MARCELO EIRAS ◽  
ALEXANDRE L. R. CHAVES ◽  
ADDOLORATA COLARICCIO ◽  
RICARDO HARAKAVA ◽  
JANSEN DE ARAUJO ◽  
...  

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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