First Report of Iris yellow spot virus in Onion in Hawaii

Onion (Allium spp.) production in Hawaii is mostly comprised of green onion and the locally prized sweet bulb onions (Allium cepa L.) that include short- and medium-day cultivars. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an important constraint to bulb and seed onion production in many onion-growing regions of the continental United States and the world (3). In June 2010, straw-colored, diamond-shaped lesions with occasional green islands were observed on leaves of sweet onion ‘Linda Vista’ in an insecticide trial on Maui for onion thrips (Thrips tabaci) control. Collapse and lodging occurred when lesions on leaves were severe. Seven bulbs with green leaves exhibiting lesions were collected from this onion field in the Pulehu Region of the lower Kula District on Maui. Leaf samples that included a lesion or were within 1 cm of a lesion were found to be positive in indirect ELISA with IYSV-specific polyclonal antisera (2). A405nm readings after 1 h ranged from 0.263 to 2.067 for positive samples and 0.055 to 0.073 for healthy onion controls. Four samples that were prepared from leaf tissue several centimeters away from a lesion tested negative in ELISA. Such uneven virus distribution in the plants has been previously reported (4). In July 2010, symptomatic sweet onion from a commercial farm in upper Kula, Maui at the 1,060 to 1,220 m (3,500 to 4,000 foot) elevation tested positive for IYSV by ELISA. Green onion samples collected from a commercial farm in Omaopio, Maui, located approximately 0.8 km (0.5 mile) north of Pulehu, have tested negative, suggesting distribution may be limited at this time. RNA was isolated from leaf tissue from the seven ‘Linda Vista’ sweet onions collected from the Maui insecticide trial. Reverse transcription (RT)-PCR with forward and complementary primers 5′-CTCTTAAACACATTTAACAAGCAC-3′ and 5′-TAAAACAAACATTCAAACAA-3′ flanking the nucleocapsid (N) gene encoded by the small RNA of IYSV was conducted as previously described (1). Amplicons approximately 1.1 kb long were obtained from all seven symptomatic onion samples but not from healthy samples or water controls. Sequencing of selected amplicons confirmed IYSV infection. Three sequence variants (GenBank Accession Nos. HM776014–HM776016) were identified from two RT-PCR reactions. Phylogenetic analyses of the three sequence variants with the neighbor-joining procedure available through NCBI-BLASTn Tree View showed that the highest nucleotide identities of 97 to 98% were shared with IYSV isolates from New Zealand (EU477515), Nevada (FJ713699), and northern California (FJ713700). Phylogenetic analyses with the N-gene showed the sequences from Hawaii are most closely related to isolates from the western United States, Texas, and New Zealand. To date, to our knowledge, IYSV has not been detected on the islands of Kauai, Oahu, Molokai, or Hawaii. The distribution and economic consequences of this disease to Hawaii's onion production are under investigation. References: (1) H. R. Pappu et al. Arch Virol. 151:1015, 2006. (2) H. R. Pappu et al. Plant Dis. 92:588, 2008. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) T. N. Smith et al. Plant Dis. 90:729, 2006.

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First Report of Iris yellow spot virus on Onion in New York

Plant Disease ◽

10.1094/pdis-91-3-0327a ◽

2007 ◽

Vol 91 (3) ◽

pp. 327-327 ◽

Cited By ~ 13

Author(s):

C. A. Hoepting ◽

H. F. Schwartz ◽

H. R. Pappu

Keyword(s):

United States ◽

New York ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Nucleotide Sequence Analysis ◽

Rt Pcr ◽

Onion Thrips ◽

Spot Virus ◽

First Report

Iris yellow spot virus (IYSV [family Bunyaviridae, genus Tospovirus]), a potentially devastating disease of onion vectored by onion thrips (Thrips tabaci Lindeman), has been reported from most states in the western United States where significant onion production occurs, with the most recent report from Texas (1). In June 2006, volunteer onion (Allium cepa) plants in Orleans County, New York (Elba muckland) were found to have symptoms indicative of IYSV infection. The scapes (seed stalks) of the volunteer onions found at the edge of a cull pile from a 2005 onion crop exhibited diamond-shaped lesions, each with a distinct green center and a double yellow border. Approximately 25 of 100 plants of red and yellow onion cultivars exhibited characteristic IYSV lesions. The cull pile was composed primarily of locally grown onions, although a few of the bulbs were grown from imported bare-root transplants imported from Arizona. Symptomatic plants tested positive for IYSV using IYSV-specific antiserum from Agdia Inc. (Elkhart, IN) in a double-antibody sandwich-ELISA. The presence of IYSV was verified by reverse transcription (RT)-PCR using primers derived from the small RNA of IYSV (S-RNA). The primers flanked the IYSV nucleocapsid (N) gene (5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′ (3). RT-PCR assays produced a PCR amplicon of expected size (approximately 1.2 kb) and the product was cloned and sequenced. Nucleotide sequence analysis confirmed the identity of the amplicon as that of the IYSV S-RNA. Sequence comparisons showed 95 to 98% identity with known IYSV N gene sequences available in GenBank. The virus is poorly transmitted to onion by mechanical inoculation and we did not have access to a noninfested colony of the onion thrips vector to transfer the virus from these samples to noninfected onions. No asymptomatic plants were tested. Among the onion-growing states in the eastern United States, IYSV has previously only been reported from Georgia (2). To our knowledge, this is the first report of IYSV in New York and the greater northeastern United States. The finding of this disease in New York confirms further spread of the virus within North America and the need for research to develop more effective management options to reduce the impact of IYSV on onion crops. References: (1) M. Miller et al. Plant Dis. 90:1359, 2006. (2) S. W. Mullis et al. Plant Dis. 90:377, 2006. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006.

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First Report of Iris yellow spot virus in Commercial Leek (Allium porrum) in the United States

Plant Disease ◽

10.1094/pd-91-0113b ◽

2007 ◽

Vol 91 (1) ◽

pp. 113-113 ◽

Cited By ~ 5

Author(s):

H. F. Schwartz ◽

K. Otto ◽

H. R. Pappu

Keyword(s):

United States ◽

The United States ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Allium Porrum ◽

Rt Pcr ◽

Spot Virus ◽

Disease Symptoms ◽

First Report

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) has a wide host range, with onion (Allium cepa L.) being one of the most economically important hosts. IYSV has been widely reported from this species throughout most onion-production regions of the United States and many areas of the world in recent years. A relative of onion, leek (Allium porrum L.), has been reported to be a host of IYSV in countries such as the Netherlands, Reunion Island, and Australia (1,4). A related tospovirus, Tomato spotted wilt virus (TSWV), was recently reported causing necrotic lesions and extended bleaching of leaf tips of leek in Georgia (2). In September of 2006, disease symptoms suspected to be caused by IYSV were observed on central and outer leaves of plants in a 2.6-ha section of commercial leeks being grown from seed (cvs. Tadorna and King Richard). The leek plants were adjacent to a 3.1-ha section of seeded onion (cv. Exacta) that had been harvested 2 weeks earlier. Twenty-five to thirty percent of unharvested onion plants next to the leek section also exhibited IYSV-type disease symptoms generally on the central leaves. Both Allium spp. were seeded 5 months earlier and grown under certified organic, pivot-irrigated conditions in Larimer County in northern Colorado. Disease symptoms on leek and onion leaves appeared as dry, white-to-straw-colored, spindle- or diamond-shaped lesions that ranged in size from 5 to 10 × 25 to 50 mm or larger depending on lesion age. Lesion centers, especially on leek, often had green centers with concentric rings of alternating green and straw-colored tissue. Green tissue near necrotic lesions of a single symptomatic leaf from 10 plants each of leek and onion was sampled and analyzed using a double-antibody sandwich (DAS)-ELISA (Agdia, Inc., Elkhart, IN). Five of ten leek and nine of ten onion samples were positive for IYSV. Using reverse transcription (RT)-PCR and primers specific to the small RNA of IYSV (5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′), the complete nucleocapsid (N) gene was amplified from symptomatic leek plants and then sequenced (3). Comparisons with IYSV N gene sequences available in the GenBank confirmed the identity of the virus as IYSV. Leek samples were negative for TSWV when tested by RT-PCR with TSWV-specific primers. In addition, three specimens of the presumed thrips vector recovered from five IYSV-infected leek plants were identified as Thrips tabaci (L. A. Mahaffey and W. S. Cranshaw, personal communication). Earlier in the season, T. tabaci was observed in the nearby planting of onion that also exhibited IYSV in September. To our knowledge, this is the first report of natural infection of commercial leek with IYSV in the United States. The incidence of plants (25 to 30%) with foliar lesions on multiple leaves and stunting of 5% of infected plants in both leek cultivars suggests that IYSV could seriously reduce leek stem development and marketability. References: (1) I. Cortes et al. Phytopathology 88:1276, 1998. (2) C. Nischwitz et al. Plant Dis. 90:525, 2006. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (4) T. N. Smith et al. Plant Dis. 90:729, 2006.

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Outbreak of Iris yellow spot virus in Onion Seed Crops in Central Oregon

Plant Disease ◽

10.1094/pd-89-0105c ◽

2005 ◽

Vol 89 (1) ◽

pp. 105-105 ◽

Cited By ~ 14

Author(s):

F. J. Crowe ◽

H. R. Pappu

Keyword(s):

United States ◽

The United States ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Rt Pcr ◽

Spot Virus ◽

Seed Crop ◽

Onion Seed ◽

Seed Crops

Iris yellow spot virus (IYSV) of the genus Tospovirus, family Bunyaviridae is considered an emerging or reemerging pathogen affecting onions in the United States. The virus has been endemic to the Treasure Valley of southern Idaho for more than a decade (4). Reports of its further spread came from several states in the region, most recently from New Mexico and Washington (1,3). During the 2004 growing season, a few onion seed crops near Madras (Jefferson County) in central Oregon showed symptoms suggestive of IYSV infection, including characteristic diamond-shaped scape lesions (2). By July, scapes in one-half of a 4-ha field were 100% symptomatic and 95% lodged, leading to nearly total crop failure; in the other half, scapes were 30 to 40% symptomatic and 15% lodged, with symptoms and lodging increasing weekly at 8 weeks before harvest. The half of this crop with greater incidence was immediately adjacent to a field where very limited IYSV-like symptoms were noticed in a 2002–2003 onion seed crop that was harvested in mid-August 2003, after the highly symptomatic 2003–2004 onion seed crop was planted next to it in early July 2003. Both crops were planted from true seed. In another onion seed crop located 1,000 m away, IYSV-like symptoms were abundant around the field edges in July and through the field in August 2004, with approximately 5% lodging by mid-August. A small number of plants with IYSV-like symptoms were present in a few more distant fields, but not in most onion seed fields in central Oregon. Symptomatic plants were collected and tested in the laboratory for confirmation of IYSV infection. IYSV was confirmed using enzyme-linked immunosorbent assay (ELISA) with a commercially available antiserum (Agdia Inc., Elkhart, IN). Total nucleic acids were extracted, and using primers specific to the nucleocapsid (N) gene of IYSV (3), reverse transcription-polymerase chain reaction (RT-PCR) was done. RT-PCR gave DNA amplicons of the expected size. The DNA amplicons were cloned and sequenced. Nucleotide sequence comparisons with known IYSV N gene sequences confirmed virus identity. The rapid spread of IYSV in the Pacific Northwest and its severity of incidence often leading to 100% incidence is a cause for concern for onion growers and industry. Efforts to identify management practices to reduce its impact have to be undertaken on a regional basis because of its widespread occurrence across several states in the northwestern United States. References: (1) R. Creamer et al. Plant Dis. 88:1049, 2004 (2) L. J. du Toit et al. APSnet image of the week. On-line publication: http://apsnet.org/online/archive/ 2003/IW000030.asp , 2003. (3) L. J. du Toit et al. Plant Dis. 88:222, 2004. (4) J. M. Hall et al. Plant Dis. 77:952, 1993.

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First Report of Iris yellow spot virus on Onion (Allium cepa) in Texas

Plant Disease ◽

10.1094/pd-90-1359b ◽

2006 ◽

Vol 90 (10) ◽

pp. 1359-1359 ◽

Cited By ~ 7

Author(s):

M. E. Miller ◽

R. R. Saldana ◽

M. C. Black ◽

H. R. Pappu

Keyword(s):

Disease Incidence ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Nucleotide Sequence Analysis ◽

Rt Pcr ◽

Onion Thrips ◽

Spot Virus ◽

Das Elisa ◽

Bulb Yield

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) has emerged as a potentially devastating and widespread virus of onion. IYSV was first reported in the United States from Idaho in 1993 and has since spread to many of the onion-producing areas (1). In South America, the most recent reports of the virus on onion were from Peru and Chile (2,4). In 2005, onion plants in Uvalde County, Texas exhibited necrotic lesions on leaves typical of IYSV and disease incidence approached 100% in some fields with yield loss and quality problems. Five of six plants tested were positive for IYSV with double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA; Agdia Inc., Elkhart, IN). In 2006, similar lesions were observed on onion plants in Uvalde County and approximately 400 km south in Hidalgo and Cameron counties. Infection points generally started as a single plant near the edge of fields and spread to plants in a 3- to 4-m area after 1 to 2 weeks. Early-season disease incidence was low in onions grown for bulbs and transplants, <10% in 2006. Disease incidence increased in some fields until the crop was harvested. Leaves of symptomatic plants were tested for IYSV and Tomato spotted wilt virus (TSWV) using DAS-ELISA, and 18 of 23 samples from the Hidalgo County area and 12 of 21 samples from the Uvalde County area were positive for IYSV. All samples tested for TSWV from these counties were negative. Virus infection in some ELISA-positive plants was verified by reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from the small RNA of IYSV. The primers flanked the IYSV nucleocapsid (N) gene (5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′ (3). RT-PCR gave a PCR product of expected size (approximately 1.2 kb). The DNA amplicon was cloned and sequenced (GenBank Accession No. DQ658242). Nucleotide sequence analysis confirmed the identity of the amplicon as that of IYSV N gene and sequence comparisons with known IYSV N gene sequences showed 95 to 98% sequence identity. The primary vector of IYSV, onion thrips (Thrips tabaci), is a widespread and destructive pest of onion in south Texas. The year-to-year incidence of IYSV and the severity of the disease will probably depend on the onion thrips population levels. Bulb yield reduction could be severe during years with high thrips populations. More research is needed to determine the impact of IYSV on bulb yield in Texas, the relationship between IYSV incidence and T. tabaci population levels, and oversummering hosts. To our knowledge, this is the first known report of IYSV in Texas. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004, (2) S. W. Mullis et al. Plant Dis. 90:377, 2006, (3) H. Pappu et al. Arch. Virol. 151:1015, 2006. (4) M. Rosales et al. Plant Dis. 89:1245, 2005.

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Susceptibility of Onion Relatives (Allium spp.) to Iris yellow spot virus

Plant Disease ◽

10.1094/pdis-11-10-0819 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1319-1319 ◽

Cited By ~ 1

Author(s):

C. S. Cramer ◽

S. Bag ◽

H. F. Schwartz ◽

H. R. Pappu

Keyword(s):

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Rt Pcr ◽

Sources Of Resistance ◽

Spot Virus ◽

Allium Species ◽

Elisa Kit ◽

Important Constraint ◽

Allium Cepa L

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is becoming an increasingly important constraint to the production of bulb and seed onions (Allium cepa L.) in many onion-growing regions of the continental United States and the world (4). During an evaluation of onion germplasm for susceptibility to IYSV, six other Allium species (A. altaicum, A. galanthum, A. roylei, A. schoenoprasum, A. tuberosum, and A. vavilovii) were also evaluated under natural field conditions. In July 2010, symptoms suggestive of IYSV infection (straw-colored necrotic lesions) were observed on leaves of these Allium spp. in experimental plots in Las Cruces, NM. IYSV was detected in symptomatic leaves of A. altaicum, A. vavilovii, A. tuberosum, A. schoenoprasum and A. roylei with a commercially available ELISA kit (Agdia Inc., Elkhart, IN). IYSV infection was confirmed by reverse transcription (RT)-PCR with forward and complementary primers 5′-CTCTTAAACACATTTAACAAGCAC-3′ and 5′-TAAAACAAACATTCAAACAA-3′ flanking the nucleocapsid (N) gene encoded by the small RNA of IYSV as previously described (1,3). Amplicons, approximately 1.1 kb long, were obtained from all symptomatic Allium spp. samples but not from healthy samples or water controls. Sequencing of selected amplicons confirmed IYSV infection. The highest nucleotide identity of 98% was shared with IYSV isolates from Japan (GenBank Accession No. AB180921). A. altaicum, A. vavilovii, and A. pskemense were previously reported from Washington to be susceptible to IYSV (2). Current findings expand the list of Allium spp. that are susceptible to IYSV and underscores the need for continued screening of other members of the genus to find sources of resistance to IYSV. References: (1) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (2) H. R. Pappu et al. Plant Dis. 90:378, 2006. (3) H. R. Pappu et al. Plant Dis. 92:588, 2008. (4) H. R. Pappu et al. Virus Res. 141:219, 2009.

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First Report of Natural Infection of Garlic with Iris yellow spot virus in the United States

Plant Disease ◽

10.1094/pdis-93-8-0839a ◽

2009 ◽

Vol 93 (8) ◽

pp. 839-839 ◽

Cited By ~ 5

Author(s):

S. Bag ◽

P. Rogers ◽

R. Watson ◽

H. R. Pappu

Keyword(s):

United States ◽

Natural Infection ◽

Sandwich Elisa ◽

The United States ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Spot Virus ◽

Sequence Comparisons ◽

First Report

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an important constraint to onion bulb and seed production in several onion-growing regions of the United States (1,3). While garlic (Allium sativum) was reported to be infected with IYSV in Réunion Island (4), there have been no confirmed reports of natural infection of garlic in the United States. Garlic plants showing near-diamond-shaped lesions were found in August of 2008 in Marion County, Oregon. The 0.4046-ha (1-acre) field plot consisted of various true-seeded garlic varieties and was adjacent to three onion fields that showed IYSV symptoms. Symptoms were observed on 5% of the garlic plants with most of the symptomatic plants displaying small and diffuse straw-colored spots. Seven of these symptomatic plants were selected for testing. Of these, two showed characteristic diamond-shaped, elongated, straw-colored lesions on garlic scapes. However, the lesions were more diffuse with less-defined edges compared with the characteristic diamond-shaped lesions that are often associated with IYSV infection (1). All symptomatic plants were positive for IYSV by double-antibody sandwich-ELISA with a commercially available kit (Agdia Inc., Elkhart, IN). To verify IYSV infection, total nucleic acid extracts from the symptomatic parts of the leaves were prepared and tested for the presence of IYSV by reverse transcription (RT)-PCR with primers 5′-TAAAACAAACATTCAAACAA-3′ and 5′-CTCTTAAACACATTTAACAAGCAC-3′, which flank the nucleocapsid (N) gene coded by the small RNA of IYSV (2). An approximate 1.1-kb amplicon was obtained from all symptomatic plants and cloned and sequenced. Nucleotide sequence comparisons using BLAST showed that a consensus of three clones derived from the amplicon from garlic (No. FJ514257) was 85 to 99% identical with IYSV sequences available in GenBank (Nos. AF001387, AB180918, and AB286063), confirming the identity of IYSV. To our knowledge, this is the first report of natural infection of IYSV infection of garlic in the United States. Additional surveys and testing are needed to obtain a better understanding of IYSV incidence in garlic to evaluate its impact on garlic production. References: (1) D. Gent et al. Plant Dis. 90:1468, 2006. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006.

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Iris yellow spot virus in Onion in Nevada and Northern California

Plant Disease ◽

10.1094/pdis-93-6-0674c ◽

2009 ◽

Vol 93 (6) ◽

pp. 674-674 ◽

Cited By ~ 3

Author(s):

S. Bag ◽

J. Singh ◽

R. M. Davis ◽

W. Chounet ◽

H. R. Pappu

Keyword(s):

United States ◽

Online Publication ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Plant Health ◽

N Gene ◽

Northern California ◽

Spot Virus ◽

Health Progress ◽

Seed Crops

The disease caused by thrips-transmitted Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) has become a major constraint to bulb and seed onion crops in several parts of the country and the world (1,3). As part of an ongoing survey for IYSV incidence in onion in the western United States, commercial fields in Lyon County, Nevada and several commercial fields in the northern Californian counties of Colusa, San Benito, Sutter, and Yolo were surveyed during the summer of 2008. Symptomatic plants were found widespread in northern California, especially in seed-production fields. In Lyon County, NV, symptoms were observed only on volunteer onions in one commercial field. Symptoms on leaves and scapes included characteristic diamond-shaped lesions with or without green islands. Four samples from Nevada and fourteen from northern California were tested by double-antibody sandwich (DAS)-ELISA using a commercially available kit (Agdia Inc., Elkhart, IN). All tested samples were found positive in ELISA. IYSV infection was verified by reverse transcription (RT)-PCR. Total nucleic acids were prepared from symptomatic tissue, and primers specific to the small (S) RNA of IYSV were used to amplify an approximate 1.2-kb region of the S-RNA. This region included the complete nucleoprotein (N) gene (2). The amplicons from one sample each from Nevada and northern California were sequenced (GenBank Accession Nos. FJ713699 and FJ713700, respectively). Sequence analysis showed that the amplicons contained a single open reading frame of 822 bp, coding for a 273-amino acid N protein, and the gene shared 96 to 98% identity with known IYSV N gene sequences. To our knowledge, this is the first report of IYSV in onion in Nevada. In California, outbreaks of IYSV had been reported earlier in Imperial Valley and Antelope Valley in southern California (4), and the disease has been increasing in incidence in bulb and seed crops in northern California, as well. California and Nevada are major onion-producing states in the United States and regular surveys to determine the incidence and impact on yield are needed to develop an integrated disease management program. References: (1) D. H. Gent et al. Plant Dis. 90:1468, 2006. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu and M. E. Matheron. Online publication. doi:10.1094/PHP-2008-0711-01-BR. Plant Health Progress, 2008. (4) G. J. Poole et al. Online publication. doi:10.1094/PHP-2007-0508-01-BR. Plant Health Progress, 2007.

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First Report of Iris yellow spot virus in Commercial Leek (Allium porrum) in Spain

Plant Disease ◽

10.1094/pdis-91-10-1365b ◽

2007 ◽

Vol 91 (10) ◽

pp. 1365-1365 ◽

Cited By ~ 5

Author(s):

C. Córdoba-Sellés ◽

C. Cebrián-Mico ◽

A. Alfaro-Fernández ◽

M. J. Muñoz-Yerbes ◽

C. Jordá-Gutiérrez

Keyword(s):

Natural Infection ◽

The United States ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Nucleotide Sequence Analysis ◽

Allium Porrum ◽

Rt Pcr ◽

Spot Virus ◽

First Report

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) has a wide host range, with onion (Allium cepa L.) being one of the most economically important hosts. The first report of IYSV in Spain was from Albacete in 2003 (1) followed by the Canary Islands in 2005. In November of 2006, disease symptoms suspected to be caused by IYSV were observed on the central and outer leaves of commercial leeks plants (cvs. Asthow, Edison, and Shelton) from Alicante, Spain. Symptoms consisted of dry, white-to-straw-colored, spindle-shaped, irregular chlorotic and necrotic lesions on the leaves. Tissue from symptomatic leaves was sampled and analyzed by a double-antibody sandwich (DAS)-ELISA with specific polyclonal antibodies against Onion yellow dwarf virus (OYDV), Leek yellow stripe virus (LYSV) (Biorad Phyto-Diagnostics, Marnes-La Coquette, France), IYSV, and Tomato spotted wilt virus (TSWV) (Loewe Biochemica, Sauerlach, Germany). Five of seven leek samples belonging to the three cultivars tested were positive for IYSV. All samples were negative for the other viruses tested. The presence of IYSV was verified in the positive samples by reverse transcription (RT)-PCR using primers derived from the nucleocapsid (N) gene of IYSV (1). RT-PCR gave a PCR amplicon of expected size (approximately 790 bp) from symptomatic leek plants. The product of one of the positive leek samples was purified and sequenced (GenBank Accession No. EF427447). Nucleotide sequence analysis confirmed the identity of the amplicon as that of the IYSV N gene. Sequence comparisons showed 99% identity with the sequence of the IYSV Spanish isolate available in GenBank (Accession No. EF419888). Thrips tabaci is the primary vector of IYSV. Although the vector is present in Spain, the efficiency of the Mediterranean ecotype in transmitting the virus is not known. Leek has been reported to be a host of IYSV in countries such as the Netherlands, Reunion Island, Australia, and the United States (2). To our knowledge, this is the first report of natural infection of leek with IYSV in Spain. References: (1) C. Córdoba-Sellés et al. Plant Dis. 89:1243, 2005. (2) H. F. Schwartz et al. Plant Dis. 91:113, 2007.

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Identification of New Alternative Weed Hosts for Iris yellow spot virus in the Pacific Northwest

Plant Disease ◽

10.1094/pdis-91-12-1683b ◽

2007 ◽

Vol 91 (12) ◽

pp. 1683-1683 ◽

Cited By ~ 14

Author(s):

R. K. Sampangi ◽

S. K. Mohan ◽

H. R. Pappu

Keyword(s):

Plant Species ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Onion Bulb ◽

N Gene ◽

Rt Pcr ◽

Weed Species ◽

Spot Virus ◽

Viral Pathogen ◽

Redroot Pigweed

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of onion bulb and seed crops in several parts of the United States and the world (1). IYSV is primarily transmitted by onion thrips (Thrips tabaci) and there is no evidence of seed transmission (1). However, susceptible cultivated and weed species could serve as reservoirs of inoculum from which thrips could acquire the virus to introduce and spread it in onion fields. Samples from asymptomatic and symptomatic volunteer onion plants in some of the commonly cultivated crops in the region (corn, wheat, grapes, mint, carrot, alfalfa, and sugar beets) and several common weeds in and around onion bulb and seed fields with a history of IYSV in Idaho and Washington were collected during the months of July, August, September and October of 2006. More than 175 samples from 35 plant species were analyzed for IYSV by a commercially available ELISA kit (Agdia Inc., Elkhart, IN). With the exception of a few volunteer onions, none of the other plant species had any symptoms of virus infection. Symptoms on volunteer onions included characteristic diamond-shaped lesions. To confirm the presence of IYSV in the ELISA-positive samples, total nucleic acids were extracted (2) and used in a reverse transcription (RT)-PCR assay (3). The primer pair consisted of 5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′. This primer pair flanks the nucleocapsid (N) gene of IYSV and generates an approximate 1.2-kb amplicon (3) that includes the complete N gene. An amplicon of expected size was obtained from each IYSV-positive sample. The amplicons were cloned and sequenced. There was a 95% sequence identity with known IYSV sequences. While several weed species gave ELISA values that suggested the presence of IYSV, results of RT-PCR assays failed to confirm the presence of the virus. This discrepancy between ELISA and RT-PCR results could be due to nonspecific reaction in ELISA (4) or difficulty associated with obtaining RT-PCR-quality templates for amplification. Only volunteer onions and the following weeds tested positive for IYSV by ELISA and RT-PCR: redroot pigweed (Amaranthus retroflexus), puncturevine (Tribulus terrestris), kochia (Kochia scoparia), prickly lettuce (Lactuca serriola), and common lambsquarters (Chenopodium album). Of these, redroot pigweed was recently reported to be ELISA-positive for IYSV (1). This information on the wider natural host range of IYSV, including potential alternative hosts that could serve as virus reservoirs, is useful for a better understanding of the disease epidemiology and in developing an integrated management strategy for reducing the impact of this disease. References: (1) D. Gent et al. Plant Dis. 90:1468, 2006. (2) H. R. Pappu et al. HortScience 40:697, 2005. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (4) T. N. Smith et al. Plant Dis. 90:729, 2006.

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First Report of Iris yellow spot virus Infecting Onion in Bosnia and Herzegovina

Plant Disease ◽

10.1094/pdis-09-12-0893-pdn ◽

2013 ◽

Vol 97 (3) ◽

pp. 430-430 ◽

Cited By ~ 1

Author(s):

V. Trkulja ◽

J. Mihić Salapura ◽

D. Kovačić ◽

I. Stanković ◽

A. Bulajić ◽

...

Keyword(s):

Bosnia And Herzegovina ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Post Inoculation ◽

Rt Pcr ◽

Spot Virus ◽

Susceptible Host ◽

First Report ◽

Negative Controls

In July 2012, a survey was conducted to determine the presence of tospoviruses in Bosnia and Herzegovina, symptoms resembling those caused by Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) were observed in an onion (Allium cepa) seed crop in the Gornji Karajzovci locality (Region of Banja Luka). Symptoms included chlorotic to necrotic, straw-colored, spindle- and diamond-shaped lesions, variable in size and randomly distributed on the leaves and particularly on the scapes. Later the lesions enlarged and coalesced, causing scape breakage. Affected plants occurred throughout the field and disease incidence was estimated at 20%. Symptomatic plants were sampled and assayed by double-antibody sandwich (DAS)-ELISA test using commercial polyclonal antisera (Bioreba AG, Reinach, Switzerland) against IYSV and two other tospoviruses, Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV). Commercial positive and negative controls were included in each test. IYSV was detected serologically in 19 of 20 screened samples and none of the samples tested positive for TSWV or INSV. The virus was mechanically transmitted from an ELISA-positive sample (302-12) to five of each Petunia × hybrida and Nicotiana benthamiana using chilled 0.01 M phosphate buffer (pH 7) containing 0.1% sodium sulfite (1). All inoculated P. × hybrida showed local necrotic spots, while N. benthamiana developed mild mosaic 4 and 10 days post-inoculation, respectively. However, difficulties were encountered in reproducing the disease symptoms on mechanically inoculated onion plants corroborating a previous study (2). Serological findings were verified with reverse transcription (RT)-PCR. Total RNAs from all naturally infected onion plants as well as mechanically infected N. benthamiana plants were extracted with the RNease Plant Mini Kit (Qiagen, Hilden, Germany). RT-PCR was performed with One-Step RT-PCR Kit (Qiagen) using IYSV-specific primers IYSV56U/IYSV917L (3), designed to amplify an 896-bp fragment of the S RNA which includes whole nucleocapsid (N) gene. Total RNAs from Serbian IYSV isolate from onion (GenBank Accession No. EU586203) and from healthy onion plants were used as positive and negative controls, respectively. An amplicon of the expected size was obtained from each of the plants assayed as well as from positive control, but not from the negative control. The amplified products derived from onion isolate 302-12 was purified (QIAquick PCR Purification Kit, Qiagen), sequenced directly (JX861126), and compared with known IYSV isolates. Sequence analysis of the complete N gene, conducted with MEGA5 software (4), revealed the highest nucleotide identity of 99.5% (100% amino acid identity) with IYSV onion isolate (DQ658242) from Texas. To our knowledge, this is the first report of IYSV in Bosnia and Herzegovina. Onion is an important and traditionally grown vegetable crop in Bosnia and Herzegovina and the presence of IYSV could represent an important constraint to onion and other susceptible host production. The discovery of IYSV on onion should prompt more detailed surveys, thorough inspections and subsequent testing to establish the distribution and incidence of IYSV in Bosnia and Herzegovina. References: (1) A. Kritzman et al. Plant Dis. 85:838, 2001. (2) L. Pozzer et al. Plant Dis. 83:345, 1999. (3) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

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