Increase of Tospoviral Diversity in Brazil with the Identification of Two New Tospovirus Species, One from Chrysanthemum and One from Zucchini

During a survey conducted in several different regions of Brazil, two unique tospoviruses were isolated and characterized, one from chrysanthemum and the other from zucchini. The chrysanthemum virus displayed a broad host range, whereas the virus from zucchini was restricted mainly to the family Cucurbitaceae. Double-antibody sandwich-enzyme-linked immunosorbent assay and western immunoblot analyses demonstrated that both viruses were serologically distinct from all reported tospovirus species including the recently proposed peanut yellow spot virus and iris yellow spot virus (IYSV) species. The nucleotide sequences of the nucleocapsid (N) genes of both viruses contain 780 nucleotides encoding for deduced proteins of 260 amino acids. The N proteins of these two viruses displayed amino acid sequence similarities with the previously described tospovirus species ranging from 20 to 75%, but they were more closely related to each other (80%). Based on the biological and molecular features, these viruses are proposed as two new tospovirus species, designated chrysanthemum stem necrosis virus (CSNV) and zucchini lethal chlorosis virus (ZLCV). With the identification of CSNV and ZLCV, in addition to tomato spotted wilt virus, groundnut ring spot virus, tomato chlorotic spot virus, and IYSV, Brazil harbors the broadest spectrum of tospovirus species reported.

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Occurrence and Geographical Distribution of Tospovirus Species Infecting Tomato Crops in Argentina

Plant Disease ◽

10.1094/pdis.2001.85.12.1227 ◽

2001 ◽

Vol 85 (12) ◽

pp. 1227-1229 ◽

Cited By ~ 23

Author(s):

L. V. Williams ◽

P. M. López Lambertini ◽

K. Shohara ◽

E. B. Biderbost

Keyword(s):

Geographical Distribution ◽

Genetic Resistance ◽

Control Program ◽

Enzyme Linked Immunosorbent Assay ◽

Chlorotic Spot ◽

Tomato Spotted Wilt ◽

Tospovirus Species ◽

Polyclonal Antisera ◽

Rational Control ◽

Wilt Virus

“Peste negra” is a disease, caused by tospoviruses, that affects tomato crops in Argentina. Knowledge of the diversity, frequency, and distribution of different tospoviruses is essential for developing a rational control program based on genetic resistance sources. A study of the geographical distribution of tospoviruses affecting tomato crops in Argentina is presented in this paper. The areas surveyed were between the Tropic of Capricorn and 40°S and between longitude 58°W and 70°W. Tospovirus species were identified through double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), using polyclonal antisera against Tomato spotted wilt virus (TSWV), Groundnut ringspot virus (GRSV), and Tomato chlorotic spot virus (TCSV). From tomato samples that reacted positively with any of the used antisera, 63% were GRSV, 28.2% were TCSV, and 8.8% were TSWV. A differential geographical distribution of tospoviruses was determined. Every plant that tested positive for GRSV was from central and northwest Argentina, while every plant TCSV-positive was from the northeast. TSWV was found only in the Río Negro Valley region in the south of the country. The wide dispersion of GRSV may be related to the spread of Frankliniella shultzei, which transmits this virus more efficiently than other vectors.

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

Plant Disease ◽

10.1094/pdis.2004.88.11.1285b ◽

2004 ◽

Vol 88 (11) ◽

pp. 1285-1285 ◽

Cited By ~ 19

Author(s):

S. W. Mullis ◽

D. B. Langston ◽

R. D. Gitaitis ◽

J. L. Sherwood ◽

A. C. Csinos ◽

...

Keyword(s):

Tomato Spotted Wilt Virus ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Unknown Etiology ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Spot Virus ◽

Tomato Spotted Wilt ◽

Wilt Virus

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

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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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Occurrence of Tospoviruses in Ornamental and Weed Species in Markazi and Tehran Provinces in Iran

Plant Disease ◽

10.1094/pd-89-0425 ◽

2005 ◽

Vol 89 (4) ◽

pp. 425-429 ◽

Cited By ~ 51

Author(s):

T. Ghotbi ◽

N. Shahraeen ◽

S. Winter

Keyword(s):

Polyclonal Antibodies ◽

Sandwich Elisa ◽

Impatiens Necrotic Spot Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Weed Species ◽

Spot Virus ◽

Chlorotic Spot ◽

Ornamental Crops ◽

Plant Sampling ◽

Wilt Virus

Damage to agricultural crops by tospoviruses has occurred sporadically in Iran in the past; however, since 2000, outbreaks of tospoviruses have been recorded every year. The most affected ornamental crops were surveyed in two main cultivation areas in provinces of Markazi (Mahallat) and Tehran in 2000-01 and 2001-02. A few weed species also were collected. In all, 513 samples (with or without any conspicuous virus symptoms) were collected and analyzed by double- and triple-antibody sandwich enzyme-linked immunosorbent assay (ELISA) with polyclonal antibodies to Tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV), and Tomato Varamin virus (ToVV), a new Tospovirus sp. from Iran. These viruses frequently were detected in samples of many different ornamentals and often in mixed infections, whereas Iris yellow spot virus (IYSV) was detected in only four samples. ToVV also was found in weeds growing in Chrysanthemum fields and in a Cuscuta sp. Applying double-antibody sandwich ELISA, no positive reactions were found with Tomato chlorotic spot virus (TCSV). Of the total of 513 samples tested, 345 samples did not react with any Tospovirus antisera. In Tehran, INSV was identified in 21 samples (10%), IYSV in 4 samples (2%), TSWV in 16 samples (8%), and ToVV in 22 samples (11%). In Markazi province, INSV was identified in 24 samples (8%), IYSV in 1 sample (0.5%), TSWV in 40 samples (13%), and ToVV in 36 samples (12%). ToVV was found to prevail in Tehran province and TSWV in Markazi. Thrips spp. present at the plant sampling sites also were collected and identified.

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Lisianthus Leaf Necrosis: A New Disease of Lisianthus Caused by Iris yellow spot virus

Plant Disease ◽

10.1094/pdis.2000.84.11.1185 ◽

2000 ◽

Vol 84 (11) ◽

pp. 1185-1189 ◽

Cited By ~ 31

Author(s):

A. Kritzman ◽

H. Beckelman ◽

S. Alexandrov ◽

J. Cohen ◽

M. Lampel ◽

...

Keyword(s):

Polyclonal Antibodies ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Spot Virus ◽

Systemic Necrosis ◽

Nucleocapsid Gene ◽

Pcr Product ◽

Wilt Virus

Unusual viral symptoms were seen on lisianthus (Eustoma russellianum) grown in the Besor area in Israel. Symptoms included necrotic spots and rings on leaves and systemic necrosis. Preliminary analyses suggested that the disease was caused by a tospovirus. Virus particles typical of a tospovirus were observed with electron microscopy in samples taken only from symptomatic leaves. Double-antibody sandwich enzyme-linked immunosorbent assay tests of leaf sap, extracted from lisianthus and mechanically inoculated indicator plants, gave a strong positive reaction to Iris yellow spot virus (IYSV). Polyclonal antibodies prepared against IYSV enabled specific detection of the virus in crude sap from infected plants. Western blot analysis showed that IYSV was serologically distinct from Tomato spotted wilt virus (TSWV). Primers specific to the nucleocapsid gene of IYSV were used in a reverse transcription-polymerase chain reaction assay (RT-PCR) to verify the presence of IYSV. RT-PCR gave an expected PCR product of approximately 850 bp. The sequence of the cloned nucleocapsid gene confirmed the identity of IYSV, thus confirming IYSV infection of lisianthus. This is the first report of IYSV infection in dicotyledons.

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First Report of a Tospovirus on Sunflower (Helianthus annus L.) from India

Plant Disease ◽

10.1094/pdis.2000.84.12.1343b ◽

2000 ◽

Vol 84 (12) ◽

pp. 1343-1343 ◽

Cited By ~ 7

Author(s):

K. Venkata Subbaiah ◽

D. V. R. Sai Gopal ◽

M. Krishna Reddy

Keyword(s):

Coat Protein ◽

Gel Electrophoresis ◽

Tomato Spotted Wilt Virus ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Enzyme Linked Immunosorbent Assay ◽

Necrosis Virus ◽

Spot Virus ◽

Tomato Spotted Wilt ◽

Wilt Virus

Virus-like symptoms were observed on sunflower in and around Tirupati during January 1998. Infected plants exhibited severe mosaic, systemic necrosis along the stem and floral heads, leaf distortion, and ringspots on leaves. The causal virus, mechanically transmissible from sunflower to sunflower cvs. Morden, MHSF8, MHSF18, KBHS1, and Cargil, developed symptoms like those in the original plant. The virus caused chlorotic and necrotic spots on Chenopodium amaranticolor, chlorotic and necrotic rings on cowpea cv. C-152, chlorotic spots on Datura metal and Petunia hybrida, chlorotic rings and systemic infection on Gomphrena globosa, tarlike symptoms on Catharanthus roseus, and local brown lesions on Cassia tora (1). Virus was isolated from infected sunflower leaves (2), and particles in negatively stained preparations were enveloped and 80 to 90 nm in diameter. Cytopathic effects included accumulation of virus particles in the endoplasmic reticulum, the formation of viroplasm, and aggregates consisting of nonenveloped viral nucleocapsids in the cytoplasm of ultrathin sections of infected sunflower leaves. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of capsid protein resolved as a single band of 31 kD. The dsRNA isolated from infected sunflower leaves resolved as three bands of 9, 4.9, and 3 kb in 2% agarose gel electrophoresis. In enzyme-linked immunosorbent assay, the purified virus reacted with homologous and Peanut bud necrosis virus (PBNV-ICRISAT, India) antisera and not to polyclonal antibodies to Iris yellow spot virus (Netherlands), Tomato spotted wilt virus-T (Georgia), Impatiens necrotic spot virus, and several isolates of Cucumber mosaic virus (CMV-B, CMV-C, CMV-To). In western blotting analysis, the virus coat protein reacted with homologous and PBNV antisera corresponding to coat protein band of 31 kD. In reverse transcription polymerase chain reaction, the viral RNA was amplified by using primers derived from NP gene sequence of PBNV and Watermelon silver mottle virus (WSMV). Based on these properties, the virus causing sunflower mosaic followed by necrosis in India was identified as a tospovirus, which may be as a distinct isolate of sero group IV. References: (1) A. A. Brunt et al. Viruses of Plants Online. 1996. Australian National University, Canberra, 1996. (2) D. V. R. Reddy et al. Bud necrosis virus: A disease of peanut caused by Tomato spotted wilt virus. ICRISAT Inf. Bull. No. 31, 1991.

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Application of Phage Display in Selecting Tomato spotted wilt virus-Specific Single-Chain Antibodies (scFvs) for Sensitive Diagnosis in ELISA

Phytopathology ◽

10.1094/phyto.2000.90.2.183 ◽

2000 ◽

Vol 90 (2) ◽

pp. 183-190 ◽

Cited By ~ 22

Author(s):

Remko A. Griep ◽

Marcel Prins ◽

Charlotte van Twisk ◽

Hans J. H. G. Keller ◽

Randolf J. Kerschbaumer ◽

...

Keyword(s):

Phage Display ◽

Dna Sequences ◽

Tomato Spotted Wilt Virus ◽

Mottle Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Single Chain ◽

Spot Virus ◽

Single Chain Antibodies ◽

Tomato Spotted Wilt ◽

Wilt Virus

A panel of recombinant single-chain antibodies (scFvs) against structural proteins of Tomato spotted wilt virus (TSWV) was retrieved from a human combinatorial scFv antibody library using the novel phage display technique. After subcloning the encoding DNA sequences in the expression vector pSKAP/S, which allowed the scFvs to be expressed as alkaline phosphatase fusion proteins, 17 different scFv antibodies were obtained. Of these, 12 scFvs were directed against the nucleoprotein (N) and 5, putatively, against the glycoproteins (G1 and G2). Five of the N-specific antibodies cross-reacted with two other tospoviruses (Tomato chlorotic spot virus and Groundnut ringspot virus), but none recognized the more distantly related tospoviruses Impatiens necrotic spot virus, Watermelon silverleaf mottle virus, Iris yellow spot virus, or Physalis severe mottle virus. The successful use of one of the antibodies as coating and detection reagent in a double-antibody sandwich enzyme-linked immunosorbent assay showed the potential of the phage display system in obtaining antibodies for routine TSWV diagnosis.

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

Plant Disease ◽

10.1094/pdis.2003.87.1.102c ◽

2003 ◽

Vol 87 (1) ◽

pp. 102-102 ◽

Cited By ~ 3

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.

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

Plant Disease ◽

10.1094/pd-89-0340c ◽

2005 ◽

Vol 89 (3) ◽

pp. 340-340 ◽

Cited By ~ 11

Author(s):

K. L. Perry ◽

L. Miller ◽

L. Williams

Keyword(s):

New York ◽

Tomato Spotted Wilt Virus ◽

New York State ◽

Impatiens Necrotic Spot Virus ◽

Spot Virus ◽

York State ◽

Tomato Spotted Wilt ◽

In Vitro Plantlets ◽

Wilt Virus

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

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