scholarly journals First Report of Beet virus Q in Spain

Plant Disease ◽  
2006 ◽  
Vol 90 (1) ◽  
pp. 110-110 ◽  
Author(s):  
C. Rubies Autonell ◽  
C. Ratti ◽  
R. Resca ◽  
M. De Biaggi ◽  
J. Ayala García

Beet virus Q (BVQ) is a member of the genus Pomovirus that is transmitted by Polymyxa betae Keskin. Initially described as the Wierthe serotype of Beet soilborne virus (BSBV), BVQ is now considered a distinct virus species based on its genomic properties (1). BVQ is commonly found in fields where BSBV and the causal agent of rhizomania disease, Beet necrotic yellow vein virus (BNYVV), are also present. Simultaneous infection of sugar beet plants with multiple virus species could affect disease symptom expression (4). For this reason, the pathogenicity of BVQ and its role in the epidemiology of rhizomania disease remain a subject of study. During 2004, six soil samples were collected from different sites in the Castilla-La Mancha Region in Spain (Albacete and Ciudad Real provinces) where rhizomania symptoms were observed in BNYVV-tolerant sugar beet cultivars. Soil from the Hainaut Region of Belgium, infected with BNYVV, BSBV, and BVQ and supplied by Prof. C. Bragard (Unité de Phytopathologie, Université Catholique de Louvain, Belgium) was used as a positive control. Sugar beet plants (cv. Asso) were grown in the soil samples for 45 days at 24°C and then root tissue was harvested. All samples were analyzed using enzyme-linked immunosorbent assay (ELISA) with commercial BNYVV antiserum (BIOREBA AG, Reinach, Switzerland) and BSBV/BVQ antisera (IC10 and 6G2) supplied by R. Koenig (Federal Biological Research Centre for Agriculture and Forestry, Braunschweig, Germany). Total RNA extracted from sugar beet roots as previously described (3) was tested using reverse transcription-polymerase chain reaction (RT-PCR). Primers BVQ3F (5′-GTT TTC AAA CTT GCC ATC CT-3′) and BVQ3R2 (5′-CCA CAA TGG GCC AAT AGA-3′), which amplify a 690-bp fragment of the triple gene block region of BVQ RNA 3, were designed based on the published sequence (GenBank Accession No. AJ223598). The presence of BSBV and BNYVV was assayed using RT-PCR with previously described primers (2,3). BVQ was detected from plants grown in soil collected from La Roda (Albacete) in Spain and from Hainaut in Belgium. The fragments amplified from Spanish sample with BVQ3F and BVQ3R2 (GenBank Accession No. AY849375) showed 95.9% nucleotide sequence identity with the previously published sequence of BVQ (1). The La Roda BVQ isolate was mechanically transmitted to Chenopodium quinoa from infected sugar beet root tissue. BVQ was detected using RT-PCR in local lesions that appeared approximately 5 days after inoculation and subsequently spread along veins. To our knowledge, this is the first report of BVQ in soil from Spain, although it has been previously reported in Belgium, Bulgaria, France, Germany, Hungary, and the Netherlands (2). BSBV and BNYVV (type A) were detected in all six Spanish samples, as well as in the Belgian soil. References: (1) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (2) A. Meunier et al. Appl. Environ Microbiol. 69:2356, 2003. (3) C. Ratti et al. J. Virol. Methods 124:41, 2005. (4) C. Rush Annu. Rev Phytopathol 41:567, 2003.

Plant Disease ◽  
2006 ◽  
Vol 90 (10) ◽  
pp. 1363-1363 ◽  
Author(s):  
N. Borodynko

The objective of this work was to determine whether Beet virus Q (BVQ), a member of the genus Pomovirus, is present in Poland. BVQ, like Beet necrotic yellow vein virus (BNYVV), is transmitted by Polymyxa betae Keskin. Earlier, BVQ was described as the Wierthe serotype of Beet soilborne virus (BSBV). Now, on the basis of its genomic properties (2), BVQ is recognized as a distinct virus species. BVQ is often found in fields where BSBV and BNYVV are present (4). During the fall of 2005, five plants of a cultivar susceptible to rhizomania (cv. Alyssa) and five resistant to rhizomania (cv. Henrietta) were collected from a field in the Wielkopolska Region of Poland, where BSBV and BNYVV had been previously identified, and tested for BVQ (1). All samples were analyzed by a double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) with antiserum against BNYVV (Bio-Rad, Hercules, CA). Rhizomania was identified only in sugar beet samples of the susceptible variety. The same samples were then tested using a triple antibody sandwich (TAS)-ELISA with commercial antisera against BSBV/BVQ (As-0576.2) and BSBV (As-0576.1) (DSMZ, Braunschweig, Germany). Nine sugar beet plants gave positive reactions with antiserum against BSBV/BVQ and negative reactions with antiserum specific to BSBV. Total RNA extracted from roots of 10 beet samples was then tested using a multiplex reverse transcription-polymerase chain reaction (mRT-PCR) and specific primers designed to amplify a fragment of the RNA2 for BNYVV and BVQ (3). The primers specifically amplified fragments of 545 bp and 291 bp of the BNYVV and BVQ, respectively. BNYVV was detected in all five samples from susceptible sugar beet plants. The presence of BVQ was confirmed in nine of the sugar beer plants, and the RT-PCR products were sequenced. Sequence analysis of the 206-nt amplicon sequence of the Polish isolate of BVQ (GenBank Accession No. DQ309444) indicated 97% nucleotide and 94% amino acid sequence identity with the previously published sequence of BVQ (GenBank Accession No. AJ223596) (2). To my knowledge, this is the first report of the natural occurrence of BVQ on sugar beet in Poland. In Europe, it has been previously reported in Belgium, Bulgaria, France, Germany, Hungary, Italy, the Netherlands, Spain, and Sweden (3,4). References: (1) N. Borodynko et al. Plant Dis. 90:112, 2006. (2) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (3) A. Meunier et al. Appl. Environ. Microbiol. 69:2356, 2003. (4) C. Rubies Autonell et al. Plant Dis. 90:110, 2006.


Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1288-1288 ◽  
Author(s):  
A. Stas ◽  
A. Meunier ◽  
J.-F. Schmit ◽  
C. Bragard

Beet necrotic yellow vein virus (BNYVV), the causal agent of rhizomania disease on sugar beet, has been reported in Belgium for more than 16 years. Other soilborne viruses belonging to the genus Pomovirus, such as Beet soilborne virus (BSBV) (3) and Beet virus Q (BVQ) (1), are suspected pathogens of sugar beets grown in Belgium. During the 2000 growing season, more than 20 fields showing rhizomania-like and yellowing symptoms on sugar beet leaves were investigated for the presence of BVQ, BNYVV, and BSBV. All samples were checked by enzyme-linked immunosorbent assay (ELISA) using commercial BNYVV (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) and BSBV/BVQ (DSMZ, Braunschweig, Germany - AS-0576 polyclonal, AS-0576/2 MAb) antisera. RNA was extracted from sugar beet rootlets using an RNeasy extraction kit (Qiagen, Hilden, Germany), before performing a reverse transcription-polymerase chain reaction (RT-PCR) using primers (5′-GCTGGAGTATATCACCGATGAC-3′ and 5′-AAAATC TCGGATAGCATCCAAC-3′) designed to specifically amplify a 510-bp region of BVQ RNA-1. The presence of BSBV and BNYVV was also checked by RT-PCR using previously described primers (1,2). The BVQ-derived PCR product was sequenced and proved to be more than 99% identical to the Wierthe BVQ isolate nucleotide sequence. Soil transmission of BVQ was demonstrated through a bioassay using soil dilutions with quartz and sugar beet cv. Cadyx as bait. After 6 weeks, BVQ was detected by RT-PCR in bait plants. The putative vector, Polymyxa betae, was identified by lactophenol-cotton blue staining of the roots followed by microscopic examination. BVQ produces irregularly shaped local lesions that appear ≈5 days after mechanical inoculation and tend to spread along veins. BVQ was detected in six fields located in the Polders Region and Brabant Province of Belgium. BVQ was always found in sugar beet samples coinfected with BNYVV and BSBV. The economic significance of BVQ and its interaction with other viruses is not known. References: (1) R. Koenig et al. J. Gen. Virol. 79:2027, 1998. (2) M. Saito et al. Arch. Virol. 141:2163, 1996. (3) M. Verhoyen and M. Van den Bossche. Parasitica. 44:71, 1987.


Plant Disease ◽  
2005 ◽  
Vol 89 (12) ◽  
pp. 1359-1359 ◽  
Author(s):  
Sh. Farzadfar ◽  
R. Pourrahim ◽  
A. R. Golnaraghi ◽  
A. Ahoonmanesh

During the 2001 growing season, a survey was conducted to determine the incidence of Beet necrotic yellow vein virus (BNYVV), Beet soilborne virus (BSBV), and Beet virus Q (BVQ) in Iran. A total of 2,816 random and 76 samples with rhizomania were collected from 131 fields in the main sugar beet cultivation areas of 13 provinces in Iran. All samples were tested using a tissue-blot immunoassay (TBIA) with commercial BNYVV (As-0799.1/CG6-F4), BSBV (As-0576.1), and BSBV/BVQ (As-0576.2) antisera provided by S. Winter (DSMZ, Braunschweig, Germany). For randomly collected samples, the highest incidence of virus infection was found for BNYVV (52.3%), followed by BSBV (9.5%) and BVQ (1.5%). Co-infection of BNYVV with BSBV or BVQ was 6.6% and 0.9%, respectively. Infection with both BSBV and BVQ was found in 16 (0.6%) samples. In addition, 0.4% (12) of the samples was infected with all three viruses. Our results indicated the presence of BVQ in samples from 10 fields located in Azarbayejan-e-gharbi, Esfahan, Fars, Kermanshah, Khorasan, Lorestan, and Semnan provinces of Iran, with or without rhizomania-like symptoms. The presence of viruses was confirmed using reverse transcription-polymerase chain reaction (RT-PCR) of RNA from 81, 19, and 14 root samples with positive reaction in TBIA to BNYVV, BSBV, and BVQ, respectively, with previously described primers (3,4). The primers specifically amplified fragments of 501 bp, 602 bp, 399 bp, and 291 bp of the BNYVV RNAs 1 and 4, BSBV RNA-2, and BVQ RNA-1, respectively. Our results indicated that the samples tested were also positive using RT-PCR. The putative vector for BNYVV, BSBV, and BVQ, Polymyxa betae, was also detected in 161 samples (from 127 fields) by amplification of a 170-bp fragment of the P. betae repetitive EcoRI-like fragments using previously described primers (4). RT-PCR products from 72 BNYVV-positive sugar beet root samples from 58 fields that also gave positive reactions in TBIA were analyzed using single-strand conformation polymorphism (SSCP) as previously described with extracts from root beards of the susceptible sugar beet cvs. OPUS and IC1 grown in the soils infested with BNYVV types A and B (provided by A. Meunier, Unite de Phytopathologie-UCL-AGRO-BAPA, Louvain-la-Neuve, Belgium) as positive controls (3). The patterns obtained with SSCP were uniform and showed widespread occurrence of BNYVV type A in almost all provinces surveyed. The fragments obtained for BNYVV RNAs 1 and 4 of an isolate from Qazvin (BNQ1) were sequenced (GenBank Accession Nos. AY703452 and AY703455) and compared with other sequences available in GenBank using Clustal W, which revealed 99.3 and 99.6% identity with the Japanese S (D84410) and Italian type A (AF197552) isolates, respectively. The economic importance of BVQ and its interactions with other sugar beet soilborne viruses remains a matter of debate. BNYVV and BSBV have been previously reported from Iran (1,2). To our knowledge, this is the first report of the natural occurrence of BVQ in sugar beets in Iran. References: (1) Sh. Farzadfar et al. Plant Dis. 86:187, 2002. (2) K. Izadpanah et al. Iran. J. Plant Pathol. 32:155, 1996. (3) R. Koenig et al. J. Gen. Virol. 76:2051, 1995. (4) A. Meunier et al. Appl. Environ. Microbiol. 69:2356, 2003.


Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 423-423 ◽  
Author(s):  
J. A. M. Rezende ◽  
V. M. Camelo ◽  
D. Flôres ◽  
A. P. O. A. Mello ◽  
E. W. Kitajima ◽  
...  

Beet necrotic yellow vein virus (BNYVV) is an economically important pathogen of sugar beet (Beta vulgaris var. saccharifera) in several European, and Asian countries and in the United States (3). The virus is transmitted by the soil-inhabiting plasmodiophorid Polymyxa betae and causes the rhizomania disease of sugar beet. In November 2012, plants of B. vulgaris subsp. vulgaris cv. Boro (red table beet) exhibiting mainly severe characteristic root symptom of rhizomania were found in a commercial field located in the municipality of São José do Rio Pardo, State of São Paulo, Brazil. No characteristic virus-inducing foliar symptom was observed on diseased plants. The incidence of diseased plants was around 70% in the two visited crops. As the hairy root symptom is indicative of infection by BNYVV, the present study aimed to detect and identify this virus associated with the diseased plants. Preliminary leaf dip analysis by transmission electron microscopy revealed the presence of very few benyvirus-like particles. Total RNA was extracted from roots of three symptomatic plants and one asymptomatic plant according to Toth et al. (3). One-step reverse-transcription–polymerase chain reaction (RT-PCR) was performed as described by Morris et al. (2) with primers that amplify part of the coat protein gene at RNA2. The initial assumption that the hairy root symptom was associated with BNYVV infection was confirmed by the amplification of a fragment of ~500 bp from all three symptomatic samples. No amplicon was obtained from the asymptomatic control plant. Amplicons were directly sequenced, and the consensus nucleotide and deduced amino acid sequences showed 100% identity. The nucleotide sequence for one amplicon (Accession No. KM433683) was compared with other sequences deposited in GenBank. The nucleotide (468 nt) and deduced amino acid (156 aa) sequences shared 93 to 100 and 97 to 99% identity, respectively with the corresponding nucleotide and amino acid sequences for other isolates of type A of BNYVV. The virus was transmitted to three of 10 red table beet plants inoculated with contaminated soil, and infection was confirmed by nested RT-PCR, as described by Morris et al. (1), and nucleotide sequencing. This is the first report on the occurrence of BNYVV in Brazil, which certainly will affect the yield of red table beet in the producing region. Therefore, mapping of the occurrence of BNYVV in red table beet-producing areas in Brazil for containment of the spread of the virus is urgent. In the meantime, precautions should be taken to control the movement of contaminated soil and beet roots, carrots, or any vegetable grown on infested land that might introduce the virus to still virus-free regions. References: (1) J. Morris et al. J. Virol. Methods 95:163, 2001. (2) D. D. Sutic et al. Handbook of Plant Virus Diseases. CRC Press, Boca Raton, Florida, 1999. (3) I. K. Toth et al. Methods for the Detection and Quantification of Erwinia carotovora subsp. atroseptica (Pectobacterium carotovorum subsb. atrosepticum) on Potatoes: A Laboratory Manual. Scottish Crop Research Institute, Dundee, Scotland, 2002.


2003 ◽  
Vol 69 (4) ◽  
pp. 2356-2360 ◽  
Author(s):  
Alexandre Meunier ◽  
Jean-François Schmit ◽  
Arnaud Stas ◽  
Nazli Kutluk ◽  
Claude Bragard

ABSTRACT Three soilborne viruses transmitted by Polymyxa betae KESKIN in sugar beet have been described: Beet necrotic yellow vein virus (BNYVV), the agent of rhizomania, Beet soilborne virus (BSBV), and Beet virus Q (BVQ). A multiplex reverse transcription-PCR technique was developed to simultaneously detect BNYVV, BSBV, and BVQ, together with their vector, P. betae. The detection threshold of the test was up to 128 times greater than that of an enzyme-linked immunosorbent assay. Systematic association of BNYVV with one or two different pomoviruses was observed. BVQ was detected in samples from Belgium, Bulgaria, France, Germany, Hungary, Italy, Sweden, and The Netherlands but not in samples from Turkey.


Plant Disease ◽  
2007 ◽  
Vol 91 (3) ◽  
pp. 326-326 ◽  
Author(s):  
A. Kozlowska-Makulska ◽  
M. S. Szyndel ◽  
J. Syller ◽  
S. Bouzoubaa ◽  
M. Beuve ◽  
...  

Yellowing symptoms on sugar beet (Beta vulgaris L.) are caused by several viruses, especially those belonging to the genus Polerovirus of the family Luteoviridae, including Beet mild yellowing virus (BMYV) and Beet western yellows virus (BWYV), and recently, a new species, Beet chlorosis virus (BChV), was reported (2). To identify Polerovirus species occurring in beet crops in Poland and determine their molecular variability, field surveys were performed in the summer and autumn of 2005. Leaves from symptomatic beet plants were collected at 26 localities in the main commercial sugar-beet-growing areas in Poland that included the Bydgoszcz, Kutno, Lublin, Poznań, Olsztyn, and Warszawa regions. Enzyme-linked immunosorbent assay (ELISA) tests (Loewe Biochemica GmbH, Sauerlach, Germany) detected poleroviruses in 23 of 160 samples (approximately 20 samples from each field). Multiplex reverse-transcription polymerase chain reaction (RT-PCR) (1) (GE Healthcare S.A.-Amersham Velizy, France) confirmed the presence of poleroviruses in 13 of 23 samples. Nine of twenty sugar beet plants gave positive reactions with BChV-specific primers and three with primers specific to the BMYV P0 protein. Two isolates reacted only with primer sets CP+/CP, sequences that are highly conserved for all beet poleroviruses. Leaf samples collected from three plants infected with BChV were used as inoculum sources for Myzus persicae in transmission tests to suitable indicator plants including sugar beet, red beet (Beta vulgaris L. var. conditiva Alef.), and Chenopodium capitatum. All C. capitatum and beet plants were successfully infected with BChV after a 48-h acquisition access period and an inoculation access period of 3 days. Transmission was confirmed by the presence of characteristic symptoms and by ELISA. Amino acid sequences obtained from each of four purified (QIAquick PCR Purification kit, Qiagen S.A., Courtaboeuf, France) RT-PCR products (550 and 750 bp for CP and P0, respectively) were 100% identical with the CP region (GenBank Accession No. AAF89621) and 98% identical with the P0 region (GenBank Accession No. NP114360) of the French isolate of BChV. To our knowledge, this is the first report of BChV in Poland. References: (1) S. Hauser et al. J. Virol. Methods 89:11, 2000. (2) M. Stevens et al. Mol. Plant Pathol. 6:1, 2005.


Plant Disease ◽  
2003 ◽  
Vol 87 (2) ◽  
pp. 201-201 ◽  
Author(s):  
William M. Wintermantel ◽  
Teresa Crook ◽  
Ralph Fogg

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV) and vectored by the soilborne fungus Polymyxa betae Keskin, is one of the most economically damaging diseases affecting sugar beet (Beta vulgaris L.). The virus likely originated in Europe and was first identified in California in 1983 (1). It has since spread among American sugar beet production regions in spite of vigorous sanitation efforts, quarantine, and disease monitoring (3). In the fall of 2002, mature sugar beet plants exhibiting typical rhizomania root symptoms, including proliferation of hairy roots, vascular discoloration, and some root constriction (2) were found in several fields scattered throughout central and eastern Michigan. Symptomatic beets were from numerous cultivars, all susceptible to rhizomania. Two to five sugar beet root samples were collected from each field and sent to the USDA-ARS in Salinas, CA for analysis. Hairy root tissue from symptomatic plants was used for mechanical inoculation of indicator plants. Mechanical inoculation produced necrotic lesions on Chenopodium quinoa and systemic infection of Beta vulgaris ssp. macrocarpa, both typical of BNYVV and identical to control inoculations with BNYVV. Symptomatic sugar beet roots were washed and tested using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for the presence of BNYVV using standard procedures and antiserum specific for BNYVV (3). Sugar beet roots were tested individually, and samples were considered positive when absorbance values were at least three times those of greenhouse-grown healthy sugar beet controls. Samples were tested from 16 fields, with 10 confirmed positive for BNYVV. Positive samples had mean absorbance values ranging from 0.341 to 1.631 (A405nm) after 30 min. The mean healthy control value was 0.097. Fields were considered positive if one beet tested positive for BNYVV, but in most cases, all beets tested from a field were uniformly positive or uniformly negative. In addition, soil-baiting experiments were conducted on seven of the fields. Sugar beet seedlings were grown in soil mixed with equal parts of sand for 6 weeks and were subsequently tested using DAS-ELISA for BNYVV. Results matched those of the root sampling. Fields testing positive for BNYVV were widely dispersed within a 100 square mile (160 km2) area including portions of Gratiot, Saginaw, Tuscola, and Sanilac counties in the central and eastern portions of the Lower Peninsula of Michigan. The confirmation of rhizomania in sugar beet from the Great Lakes Region marks the last major American sugar beet production region to be diagnosed with rhizomania disease, nearly 20 years after its discovery in California (1). In 2002, there were approximately 185,000 acres (approximately 75,00 ha) of sugar beet grown in the Great Lakes Region, (Michigan, Ohio, and southern Ontario, Canada). The wide geographic distribution of infested fields within the Michigan growing area suggests the entire region should monitor for symptoms, increase rotation to nonhost crops, and consider planting rhizomania resistant sugar beet cultivars to infested fields. References:(1) J. E. Duffus et al. Plant Dis. 68:251, 1984. (2) J. E. Duffus. Rhizomania. Pages 29–30 in: Compendium of Beet Diseases and Insects, E. D. Whitney and J. E. Duffus eds. The American Phytopathological Society, St. Paul, MN, 1986. (3) G. C. Wisler et al. Plant Dis. 83:864, 1999.


Plant Disease ◽  
2003 ◽  
Vol 87 (1) ◽  
pp. 102-102 ◽  
Author(s):  
S. Adkins ◽  
L. Breman ◽  
C. A. Baker ◽  
S. Wilson

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.


Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1461-1461 ◽  
Author(s):  
M. J. Soule ◽  
K. C. Eastwell ◽  
R. A. Naidu

Washington State is the largest producer of juice grapes (Vitis labruscana ‘Concord’ and Vitis labrusca ‘Niagara’) and ranks second in wine grape production in the United States. Grapevine leafroll disease (GLD) is the most wide spread and economically significant virus disease in wine grapes in the state. Previous studies (2) have shown that Grapevine leafroll associated virus-3 (GLRaV-3) is the predominant virus associated with GLD. However, little is known about the incidence and economic impact of GLD on juice and table grapes. Because typical GLD symptoms may not be obvious among these cultivars, the prevalence and economic impact of GLD in Concord and Niagara, the most widely planted cultivars in Washington State, has received little attention from the grape and nursery industries. During the 2005 growing season, 32 samples from three vineyards and one nursery of ‘Concord’ and three samples from one nursery of ‘Niagara’ were collected randomly. Petiole extracts were tested by single-tube reverse transcription-polymerase chain reaction (RT-PCR; 3) with primers LC 1 (5′-CGC TAG GGC TGT GGA AGT ATT-3′) and LC 2 (5′-GTT GTC CCG GGT ACC AGA TAT-3′), specific for the heat shock protein 70 homologue (Hsp70h gene) of GLRaV-3 (GenBank Accession No. AF037268). One ‘Niagara’ nursery sample and eleven ‘Concord’ samples from the three vineyards tested positive for GLRaV-3, producing a single band of the expected size of 546 bp. The ‘Niagara’ and six of the ‘Concord’ RT-PCR products were cloned in pCR2.1 (Invitrogen Corp, Carlsbad, CA) and the sequences (GenBank Accession Nos. DQ780885, DQ780886, DQ780887, DQ780888, DQ780889, DQ780890, and DQ780891) compared with the respective sequence of a New York isolate of GLRaV-3 (GenBank Accession No. AF037268). The analysis revealed that GLRaV-3 isolates from ‘Concord’ and ‘Niagara’ share nucleotide identities of 94 to 98% and amino acid identities and similarities of 97 to 98% with the Hsp70h gene homologue of the New York isolate of GLRaV-3. Additional testing by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using antibodies specific to GLRaV-3 (BIOREBA AG, Reinach, Switzerland) further confirmed these results in the ‘Niagara’ and two of the ‘Concord’ isolates. GLRaV-3 has previously been reported in labrusca cvs. Concord and Niagara in western New York (4) and Canada (1), but to our knowledge, this is the first report of GLRaV-3 in American grapevine species in the Pacific Northwest. Because wine and juice grapes are widely grown in proximity to each other in Washington State and grape mealybug (Pseudococcus maritimus), the putative vector of GLRaV-3, is present in the state vineyards, further studies will focus on the role of American grapevine species in the epidemiology of GLD. References: (1) D. J. MacKenzie et al. Plant Dis. 80:955, 1996. (2) R. R. Martin et al. Plant Dis. 89:763, 2005. (3) A. Rowhani et al. ICGV, Extended Abstracts, 13:148, 2000. (4) W. F. Wilcox et al. Plant Dis. 82:1062, 1998.


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