scholarly journals Coriander: A New Natural Host of Groundnut Ring Spot Virus in Brazil

Plant Disease ◽  
1999 ◽  
Vol 83 (9) ◽  
pp. 878-878 ◽  
Author(s):  
M. F. Lima ◽  
A. C. de Ávila ◽  
L. J. da G. Wanderley ◽  
T. Nagata ◽  
L. J. W. da Gama
Keyword(s):  
Polyclonal Antibodies ◽  
Citrullus Lanatus ◽  
Impatiens Necrotic Spot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Natural Occurrence ◽  
Leaf Extracts ◽  
Spot Virus ◽  
Chlorotic Spot ◽  
Ring Spot ◽  
Das Elisa

Coriander plants (Coriandrum sativum L. ‘Palmeira’), showing stunting, chlorotic ring spots, necrosis, and malformation of apical leaves were observed on 50-day-old-plants in July 1998 in one seed production field at Petrolina, State of Pernambuco, Brazil, but not in nearby fields. Leaf samples were collected and tested by double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) with a panel of polyclonal antibodies made against the nucleocapsid protein (N) of tomato spotted wilt virus (TSWV), tomato chlorotic spot virus (TCSV), groundnut ring spot virus (GRSV), and impatiens necrotic spot virus (INSV) (1). All symptomatic samples reacted only with the GRSV antisera. Coriander leaf extracts from infected plants were mechanically inoculated onto potential indicator hosts. The virus induced systemic infection with vein clearing, chlorotic and necrotic spots, necrotic ring spots, mosaic, top distortion, and stunting within 21 days after inoculation on Capsicum annuum cv. Ikeda, C. chinense PI 159236, Physalis floridana, Nicandra physaloides, Nicotiana tabacum cv. TNN, N. benthamiana, Lycopersicon esculentum cv. Rutgers, Phaseolus vulgaris cv. BT2, and Gomphrena globosa. The symptomatic indicator plants tested positive for GRSV by DAS-ELISA. P. vulgaris, Chenopodium amaranthicolor, C. quinoa, and Cucurbita pepo (zucchini) cv. Caserta showed only small, necrotic, local lesions on inoculated leaves. Citrullus lanatus cv. Charleston Gray was asymptomatic. This is the first report of natural occurrence of GRSV on coriander in Brazil. Reference: (1) A. C. de Ávila et al. J. Gen. Virol. 71:2801, 1990.

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

Plant Disease ◽  
2005 ◽  
Vol 89 (4) ◽  
pp. 425-429 ◽  
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.

scholarly journals First Report of a Natural Occurrence of Tomato yellow fruit ring virus on Pepper in Iran

Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1259-1259 ◽  
Author(s):  
A. R. Golnaraghi ◽  
A. Hamedi ◽  
S. Yazdani-Khameneh ◽  
T. S. Khosroshahi
Keyword(s):  
Thrips Tabaci ◽  
Impatiens Necrotic Spot Virus ◽  
Natural Occurrence ◽  
Weed Species ◽  
Leaf Sample ◽  
Spot Virus ◽  
First Report ◽  
Chlorotic Spot ◽  
Tehran Province ◽  
Pepper Plants

Tospoviruses are considered one of the most devastating viruses in different crops and ornamentals worldwide. There have been increasing records of the natural occurrence of Tomato yellow fruit ring virus (TYFRV) in Iran (e.g., 1,2,3), a recently proposed species in the genus Tospovirus (4). During the growing seasons 2010 to 2011, surveys were conducted in pepper fields (Capsicum annum) in Tehran province, one of the main vegetable producing areas of Iran, to detect the presence of tospovirus species infecting this crop, including Groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV), Iris yellow spot virus (IYSV), Tomato chlorotic spot virus (TCSV), Tomato spotted wilt virus (TSWV), TYFRV, and Watermelon silver mottle virus (WSMoV). Overall, 14 fields were surveyed and 119 pepper leaf samples from plants showing tospovirus-like symptoms of yellow mosaic, chlorosis, and necrosis were collected. 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. Based on the results, TYFRV were found in 21 samples (17.6%) collected from five fields surveyed. None of the samples had a positive reaction in ELISA to GRSV, INSV, IYSV, TCSV, TSWV, and WSMoV. To confirm testing, six leaf samples that were found positive for TYFRV in ELISA tests were mechanically inoculated on Petunia × hybrid and Nicotiana rustica; for all the samples studied, 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. Four out of the six samples also were tested by reverse transcription (RT)-PCR technique using previously described specific primers (2). The PCR reaction, in agreement with ELISA tests, resulted in the specifically amplification of a ~1.2-kb fragment of TYFRV RNAs. Using the PCR amplification primers mentioned above, the nucleotide sequences of nucleoprotein (N) genes of two isolates, namely TY-PepT43 and TY-PepT74, were determined (GenBank Accession Nos. KC354692 and KC354693, respectively); BLAST search results confirmed the presence of TYFRV and showed high nucleotide identities (99.0%) to TY-PF36 isolate of the virus. The virus has been previously reported on potato, tomato, ornamental plants, and some weed species in Tehran Province (1,3,4). This coupled with the presence of TYFRV vector, i.e., Thrips tabaci, in the same region (1), may have resulted in the occurrence of the virus on pepper plants. To our knowledge, this is the first report of the natural occurrence of TYFRV from pepper plants in Iran. References: (1) T. Ghotbi et al. Plant Dis. 89:425, 2005. (2) A. R. Golnaraghi et al. Plant Dis. 92:1280, 2008. (3) R. Pourrahim et al. Plant Dis. 91:609, 2007. (4) S. Winter et al. Plant Pathol. 55:287, 2006.

scholarly journals First Report of Capsicum chlorosis virus Causing Yellow Stripes on Calla Lilies in Taiwan

Plant Disease ◽  
2007 ◽  
Vol 91 (9) ◽  
pp. 1201-1201 ◽  
Author(s):  
C. C. Chen ◽  
C. H. Huang ◽  
T. C. Chen ◽  
S. D. Yeh ◽  
Y. H. Cheng ◽  
...  
Keyword(s):  
Chenopodium Quinoa ◽  
Amino Acid Sequences ◽  
N Gene ◽  
Impatiens Necrotic Spot Virus ◽  
Leaf Extracts ◽  
Spot Virus ◽  
Chlorotic Spot ◽  
Calla Lily ◽  
Capsicum Chlorosis Virus ◽  
Virus Isolates

Tomato spotted wilt virus (TSWV) and Calla lily chlorotic spot virus (CCSV) are two recognized species of the Tospovirus genus in the family Bunyaviridae infecting calla lily (Zantedeschia spp.). During 2005, 15 virus isolates were collected from different calla lily plants exhibiting yellow stripes on their leaves in Ho-Li, a major calla lily-production township in Taiwan. After three successive local lesion passages on Chenopodium quinoa Willd., diseased leaf tissues individually infected by these isolates were preserved in liquid nitrogen and used for subsequent identification studies. Using the tospovirus genus-specific primers gL3637 and gL4435c designed from the L RNA, an 800-bp DNA fragment was amplified in reverse transcription-PCR from all 15 isolates. Moreover, leaf extracts of the diseased calla lilies and the C. quinoa plants inoculated with the 15 virus isolates reacted with antisera against the nucleocapsid proteins (NP) of Capsicum chlorosis virus (CaCV)-gloxinia and Watermelon silver mottle virus (WSMoV), but not to monoclonal antibodies against the NP of TSWV, CCSV, Peanut chlorotic fan-spot virus (PCFV), or Impatiens necrotic spot virus (INSV) in indirect ELISA. These results indicate that the 15 virus isolates are tospoviruses belonging to the WSMoV serogroup. Additionally, we amplified and sequenced the full-length N gene from these tospovirus isolates using primers WN2328 (5′-CCATTGGTTTGCCTCCG-3′) and WN3534 (5′-CGTCGACAGAGCAATCGAGGC-3′) designed from the S RNA of WSMoV. The deduced amino acid sequences of the N protein from these 15 tospovirus isolates showed a greater than 92% identity to that of CaCV (GenBank Accession No. NC-008301). Furthermore, results of phylogenetic analysis of the 15 isolates on the basis of amino acids sequences, both genetic distance and parsimony trees indicated that they were all genetically clustered within CaCV using INSV, TSWV, and WSMoV as outgroups. The results indicate that the virus causing yellow stripes in calla lilies is a strain of CaCV. To our knowledge, this is the first evidence that CaCV can naturally infect calla lilies and cause yellow stripe symptoms. Reference: (1) F.-H. Chu et al. Phytopathology 91:361, 2001.

scholarly journals Occurrence of Radish mosaic virus on Cauliflower and Turnip Crops in Iran

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 909-909 ◽  
Author(s):  
S. Farzadfar ◽  
R. Pourrahim ◽  
A. R. Golnaraghi ◽  
S. Jalali ◽  
A. Ahoonmanesh
Keyword(s):  
Mosaic Virus ◽  
Brassica Rapa ◽  
Polyclonal Antibodies ◽  
The United States ◽  
Plant Viruses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leaf Extracts ◽  
Chenopodium Amaranticolor ◽  
Local Lesions ◽  
Ring Spot

During the spring and summer of 2003, symptoms of mosaic, mottle, and crinkle were observed in cauliflower (Brassica oleracea) and turnip (Brassica rapa) fields in the Qazvin and Esfahan provinces of Iran, respectively. Leaf extracts of these plants, made infective by mechanical inoculation, caused necrotic local lesions on Chenopodium amaranticolor, chlorotic ring spot on Nicotiana tabacum cv. Samsun, and chlorotic local lesions followed by systemic mosaic on Brassica rapa (1). Using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and specific polyclonal antibodies (As-0120 and PV-0355) that were kindly prepared by S. Winter (DSMZ, Braunschweig, Germany), the samples were tested for the presence of Radish mosaic virus (RaMV) (family Comoviridae, genus Comovirus). ELISA results showed that the original leaf samples and inoculated indicator plants reacted positively to RaMV antibodies. RaMV has been reported in the United States, Japan, and Europe on turnip and other crucifers (1,2). To our knowledge, this is the first report of RaMV occurring in Iran. References: (1) R. N. Campbell. Radish mosaic virus. No. 121 in: Descriptions of Plant Viruses. CMI/AAB, Surrey, England, 1973. (2) D. D. Sutic et al. Handb. Plant Virus Diseases. CRC Press, Boca Raton, FL, 1999.

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

1999 ◽  
Vol 89 (9) ◽  
pp. 823-830 ◽  
Author(s):  
I. C. Bezerra ◽  
R. de O. Resende ◽  
L. Pozzer ◽  
T. Nagata ◽  
R. Kormelink ◽  
...  
Keyword(s):  
Yellow Spot ◽  
Enzyme Linked Immunosorbent Assay ◽  
Broad Host Range ◽  
Spot Virus ◽  
Chlorotic Spot ◽  
Molecular Features ◽  
Tomato Spotted Wilt ◽  
Tospovirus Species ◽  
Stem Necrosis ◽  
Wilt Virus

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.

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

2002 ◽  
Vol 27 (3) ◽  
pp. 285-291 ◽  
Author(s):  
MARCELO EIRAS ◽  
ALEXANDRE L. R. CHAVES ◽  
ADDOLORATA COLARICCIO ◽  
RICARDO HARAKAVA ◽  
JANSEN DE ARAUJO ◽  
...  
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Sao Paulo ◽  
Ringspot Virus ◽  
São Paulo ◽  
Rt Pcr ◽  
Spot Virus ◽  
Chlorotic Spot ◽  
Tomato Spotted Wilt ◽  
Wilt Virus ◽  
Das Elisa

Os tospovírus são responsáveis por perdas significativas em diversas culturas, principalmente solanáceas. No município de São José dos Campos (SP), plantas de jiló (Solanum gilo) apresentando sintomas de mosaico, bolhosidades, nanismo e queda acentuada da produção foram coletadas para análise. Visando a caracterização do agente causador dos sintomas, testes biológicos, elétrono microscópicos, sorológicos e moleculares foram realizados. Através de inoculação mecânica em plantas indicadoras das famílias Amaranthaceae, Chenopodiaceae e Solanaceae obtiveram-se resultados típicos aos esperados para tospovírus. Ao microscópio eletrônico de transmissão, observaram-se, em contrastação negativa, partículas pleomórficas com diâmetro entre 80 e 110 nm e em cortes ultra-finos partículas presentes em vesículas do retículo endoplasmático. Através de DAS-ELISA, identificou-se o Tomato chlorotic spot virus (TCSV). A partir de RNA total extraído de folhas infetadas, amplificaram-se, via RT-PCR, fragmentos correspondentes ao gene da proteína do capsídeo (cp) os quais foram seqüenciados e comparados com outros depositados no "GenBank". A homologia de nucleotídeos e aminoácidos deduzidos foi respectivamente de 99 e 95% quando comparada com seqüências de isolados de TCSV. A comparação com as outras espécies do gênero Tospovirus apresentou valores de homologia entre 72 e 84%. Estes resultados confirmam a identidade deste vírus como pertencente à espécie TCSV, que é predominante no Estado de São Paulo e importante patógeno de outras plantas cultivadas. Além disso, variedades de jiló quando inoculadas foram susceptíveis tanto ao TCSV como às espécies Tomato spotted wilt virus (TSWV) e Groundnut ringspot virus (GRSV).

scholarly journals Detection of a Severe Isolate of Impatiens Necrotic Spot Virus Infecting Lisianthus in Florida

Plant Disease ◽  
1997 ◽  
Vol 81 (11) ◽  
pp. 1334-1334 ◽  
Author(s):  
R. J. McGovern ◽  
J. E. Polston ◽  
B. K. Harbaugh
Keyword(s):  
Leaf Tissue ◽  
Impatiens Necrotic Spot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mechanical Transmission ◽  
Necrotic Spot ◽  
Spot Virus ◽  
Eustoma Grandiflorum ◽  
Systemic Necrosis ◽  
Low Incidence ◽  
Leaf Distortion

In May 1997, inclusions typical of a tospovirus were visualized by light microscopy in leaf tissue of lisianthus (Eustoma grandiflorum) exhibiting stunting, necrotic ringspots, leaf distortion, and systemic necrosis. Wilting and plant death were the final symptoms observed. Affected plants occurred at low incidence (<0.1%) in greenhouse-grown lisianthus in Manatee County, FL. Symptomatic tissue tested positive for impatiens necrotic spot virus (INSV) and negative for tomato spotted wilt virus (TSWV) with enzyme-linked immunosorbent assay (ELISA; Agdia, Elkhart, IN). Mechanical transmission of the virus to lisianthus and tomato was attempted by triturating 1 g of symptomatic leaf tissue in 7 ml of a buffer consisting of 0.01 M Tris and 0.01 M sodium sulfite, pH 7.3. Six plants of lisianthus cv. Maurine Blue and three of tomato (Lycopersicon esculentum) cv. Lanai at the second true-leaf stage were inoculated following abrasion of leaves with Carborundum. An equal number of controls were inoculated with buffer alone. Plants were maintained in a controlled environment chamber with a 12-h photoperiod, day/night temperatures of 21/16°C, and light intensity of 120 μE · s-l · m-2. Transmission rates were 100 and 0% to lisianthus and tomato, respectively. Chlorotic local lesions followed by chlorotic ringspots were observed in inoculated lisianthus leaves 4 days after inoculation. Stunting, leaf distortion, and necrotic ringspots appeared in noninoculated leaves of lisianthus plants within 3 to 4 weeks after inoculation. Buffer-inoculated lisianthus and all tomato plants remained symptomless and tested negative for INSV by ELISA. All symptomatic lisianthus tested positive for INSV by ELISA. The symptoms we observed in lisianthus due to infection by INSV were more severe than those previously reported in this host (1,2). The occurrence of such strains of INSV at high incidences could pose a significant threat for commercial lisianthus production. References: (1) M. K. Hausbeck et al. Plant Dis. 76:795, 1992. (2) H. T. Hsu and R. H. Lawson. Plant Dis. 75:292,1991.

scholarly journals First Report of Yellow Nutsedge (Cyperus esculentus) and Purple Nutsedge (C. rotundus) in Georgia Naturally Infected with Impatiens necrotic spot virus

Plant Disease ◽  
2004 ◽  
Vol 88 (7) ◽  
pp. 771-771 ◽  
Author(s):  
N. Martínez-Ochoa ◽  
S. W. Mullis ◽  
A. S. Csinos ◽  
T. M. Webster
Keyword(s):  
Southeastern United States ◽  
Impatiens Necrotic Spot Virus ◽  
Cyperus Esculentus ◽  
Necrotic Spot ◽  
Rt Pcr ◽  
Spot Virus ◽  
Black Shank ◽  
Purple Nutsedge ◽  
Yellow Nutsedge ◽  
Das Elisa

Impatiens necrotic spot virus (INSV), family Bunyaviridae, genus Tospovirus, is an emerging virus found mostly in ornamentals under greenhouse production. INSV has been detected in peanut (Arachis hypogaea L.) in Georgia and Texas (3) and recently in tobacco (Nicotiana tabacum L.) in the southeastern United States (2) but little is known about INSV distribution and impact on these crops. Noncrop plant hosts are likely to contribute to disease spread by serving as reservoirs for the virus and reproductive hosts for thrips (Frankliniella occidentalis Pergande), which transmit the virus. Yellow nutsedge, a native of North America, and purple nutsedge introduced from Eurasia, are considered serious weed problems in the southeastern United States. To date, there are no reports of natural INSV infections in these weeds. A survey was conducted at two research farms in Tift County, Georgia to determine if yellow and purple nutsedge plants were naturally infected with Tomato spotted wilt virus (TSWV) and INSV. The first field at the Black Shank Farm had been planted with flue-cured tobacco K-326 earlier in the year and fallow at the time of sampling. The second field at the Ponder Farm was planted at the time of sampling with yellow squash (Cucurbita pepo L.) and cabbage (Brassica oleracea L.). In early October 2002, 90 nutsedge plants were taken at random from each site. Leaf and root tissues of each of the nutsedge plants were tested for TSWV and INSV using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) alkaline phosphatase antisera kits (Agdia Inc., Elkhart, IN). No visible symptoms of INSV or TSWV were observed. Samples from the field at the Black Shank Farm resulted in 2 of 26 positive for INSV in purple nutsedge plants and 6 of 64 in yellow nutsedge plants. At the Ponder Farm, 3 of 12 were positive for INSV in purple nutsedge plants and 14 of 78 in yellow nutsedge plants. None of the samples in either site tested positive for TSWV. The DAS-ELISA positive samples were verified for INSV using reverse transcription-polymerase chain reaction (RT-PCR) as previously described by Dewey et al. (1). Total RNA extracts were obtained from the DAS-ELISA positive nutsedge samples using RNeasy extraction kits (Qiagen Inc., Valencia, CA). The RT-PCR was carried out with primer 1F: 5′-TCAAG(C/T) CTTC(G/T)GAA(A/G)GTGAT 3′ (1) and primer 2R: 5′-ATGAACAAAGCAAAGATTACC 3′ specific to the 3′ end of the INSV N gene open reading frame (GenBank Accession No. NC003624). DAS-ELISA negative tissues of Cyperus esculentus L. and Emilia sonchifolia (L.) DC and an E. sonchifolia DAS-ELISA positive for INSV were included in the reactions as controls. All of the DAS-ELISA positive nutsedge samples yielded an amplification product with the expected size of 298 bp when PCR products were resolved by agarose (0.7%) gel electrophoresis. The relatively high occurrence of INSV found in the sampled fields may explain the recent increase in incidence of INSV in susceptible field crops. Although yellow nutsedge is more common than purple nutsedge in North America, the potential for dispersal of INSV in both species could be significant because of the nature of nutsedge tuber survival and spreading capabilities. References: (1) R. A. Dewey et al. J. Virol. Methods 56:19, 1996. (2) N. Martínez-Ochoa et al. On-line publication. doi:10.1094/PHP-2003-0417-01-HN. Plant Health Progress, 2003. (3) S. S. Pappu et al. Plant Dis. 83:966,1999.

scholarly journals First Report of Cucurbit aphid-borne yellows virus in Iran Causing Yellows on Four Cucurbit Crops

Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 526-526 ◽  
Author(s):  
K. Bananej ◽  
C. Desbiez ◽  
C. Wipf-Scheibel ◽  
I. Vahdat ◽  
A. Kheyr-Pour ◽  
...  
Keyword(s):  
Citrullus Lanatus ◽  
Healthy Plant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Chain Reactions ◽  
Reference Isolate ◽  
Sigma Chemical ◽  
International Mycological Institute ◽  
Das Elisa

A survey was conducted from 2001 to 2004 in the major cucurbit-growing areas in Iran to reassess the relative incidence of cucurbit viruses. Severe yellowing symptoms were observed frequently on older leaves of cucurbit plants in various regions in outdoor crops, suggesting the presence of Cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus, family Luteoviridae) (1,2). Leaf samples (n = 1019) were collected from plants of melon (Cucumis melo L.), cucumber (C. sativus L.), squash (Cucurbita sp.), and watermelon (Citrullus lanatus L.) showing various virus-like symptoms (mosaic, leaf deformation, yellowing). All samples, collected from 15 provinces, were screened for the presence of CABYV by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with IgGs and alkaline phosphatase-conjugated IgGs against a CABYV reference isolate (1). Of the 1,019 samples tested, 471 were positive for CABYV using DAS-ELISA. Some of the positive samples had typical severe yellowing symptoms while symptoms in other samples were masked by mosaic or leaf deformations caused by other viruses frequently found in mixed infections (data not shown). During the entire survey, CABYV was detected by DAS-ELISA in 201 of 503 melon samples, 72 of 129 cucumber samples, 158 of 249 squash samples, and 40 of 138 watermelon samples. These results indicate that CABYV is widely distributed on four cucurbit species in the major growing areas of Iran. In order to confirm CABYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St Louis, MO) were obtained from 25 samples that were positive using DAS-ELISA originating from Khorasan (n = 4), Esfahan (n = 6), Teheran (n = 3), Hormozgan (n = 4), Azerbaiejan-E-Sharqi (n = 4), and Kerman (n = 4). Reverse transcription-polymerase chain reactions (RT-PCR) were carried out using forward (5′-CGCGTGGTTGTGG-TCAACCC-3′) and reverse (5′-CCYGCAACCGAGGAAGATCC-3′) primers designed in conserved regions of the coat protein gene according to the sequence of a CABYV reference isolate (3) and three other unpublished CABYV sequences. RT-PCR experiments yielded an expected 479-bp product similar to the fragment amplified with extracts from the reference isolate. No amplification of the product occurred from healthy plant extracts. To our knowledge, this is the first report of the occurrence of CABYV in Iran on various cucurbit species. The high frequency (46.2%) with which CABYV was detected in the samples assayed indicates that this virus is one of the most common virus infecting cucurbits in Iran. References: (1) H. Lecoq et al. Plant Pathol. 41:749, 1992 (2) M. A. Mayo and C. J. D'Arcy. Page 15 in: The Luteoviridae. H. G. Smith and H. Barker, eds. CAB International Mycological Institute, Wallingford, UK, 1999. (3) H. Guilley et al. Virology 202:1012, 1994.

scholarly journals Lamium maculatum is a Natural Host for Cucumber mosaic virus

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 150-150 ◽  
Author(s):  
R. Bešta-Gajević ◽  
A. Jerković-Mujkić ◽  
S. Pilić ◽  
I. Stanković ◽  
A. Vučurović ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Impatiens Necrotic Spot Virus ◽  
Natural Occurrence ◽  
Rt Pcr ◽  
Natural Ecosystems ◽  
New Host ◽  
Total Rna ◽  
Negative Controls ◽  
Das Elisa

Lamium maculatum L. (spotted dead-nettle) is a flowering perennial ornamental that is commonly grown as a landscape plant for an effective ground cover. In June 2010, severe mosaic accompanied by reddish brown necrosis and leaf deformation was noticed on 80% of L. maculatum growing in shade under trees and shrubs in Sarajevo (Bosnia and Herzegovina). Leaves from 10 symptomatic L. maculatum plants were sampled and analyzed by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kits (Bioreba AG, Reinach, Switzerland) against Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), and Impatiens necrotic spot virus (INSV), the most important viral pathogens of ornamental plants (1,2). Commercial positive and negative controls and extracts from healthy L. maculatum leaves were included in each assay. All samples tested negative for TSWV and INSV and positive for CMV. The virus was mechanically transmitted to test plants and young virus-free plants of L. maculatum using 0.01 M phosphate buffer (pH 7). The virus caused chlorotic local lesions on Chenopodium quinoa, while systemic mosaic was observed on Capsicum annuum ‘Rotund,’ Nicotiana rustica, N. glutinosa, N. tabacum ‘White Burley,’ and Phaseolus vulgaris ‘Top Crop.’ The virus was transmitted mechanically to L. maculatum and induced symptoms resembling those observed on the source plants. Inoculated plants were assayed by DAS-ELISA and all five inoculated plants of each species tested positive for CMV. The presence of CMV in L. maculatum as well as mechanically infected N. glutinosa plants was further confirmed by RT-PCR. Total RNA from symptomatic leaves was isolated using RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was performed with the One-Step RT-PCR Kit (Qiagen) following the manufacturer's instructions. The primer pair, CMVAu1u/CMVAu2d, that amplifies the entire coat protein (CP) gene and part of 3′- and 5′-UTRs was used for both amplification and sequencing (4). Total RNA obtained from the Serbian CMV isolate from pumpkin (GenBank Accession No. HM065510) and a healthy L. maculatum plant were used as positive and negative controls, respectively. All naturally and mechanically infected plants as well as the positive control yielded an amplicon of the expected size (850 bp). No amplicon was observed in the healthy control. The amplified product derived from isolate 3-Lam was purified (QIAquick PCR Purification Kit, Qiagen), directly sequenced in both directions and deposited in GenBank (JX436358). Sequence analysis of the CP open reading frame (657 nt), conducted with MEGA5 software, revealed that the isolate 3-Lam showed the highest nucleotide identity of 99.4% (99.1% amino acid identity) with CMV isolates from Serbia, Australia, and the USA (GQ340670, U22821, and U20668, respectively). To our knowledge, this is the first report of the natural occurrence of CMV on L. maculatum worldwide and it adds a new host to over 1,241 species (101 plant families) infected by this virus (3). This is also an important discovery for the ornamental industry since L. maculatum is commonly grown together with other ornamental hosts of CMV in nurseries and the urban environment as well as in natural ecosystems. References: (1) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (2) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (3) M. Jacquemond. Adv. Virus Res. 84:439, 2012. (4) I. Stankovic et al. Acta Virol. 55:337, 2011.

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