First Report of Iris yellow spot virus Infecting Onion in Pennsylvania

Iris yellow spot virus (IYSV; genus Tospovirus; family Bunyaviridae) is an economically important pathogen of onion. It is vectored by onion thrips (Thrips tabaci Lindeman) and causes widespread disease of onion in all major onion growing states in the western United States (1). In the eastern United States, IYSV was first reported in Georgia in 2004 (4) and then in New York in 2006 (2). In mid-July of 2010, symptomatic onion (Allium cepa) plants (cv. Candy) were found in New Holland, Pennsylvania, in Lancaster County on a small, diversified commercial farm (40.06°N, 76.06°W). Bleached, elongated lesions with tapered ends occurred on middle-aged leaves on approximately 30% of the 13,760 plants in an area approximately one tenth of an acre. Leaf tissue from five symptomatic plants tested positive for IYSV in a double-antibody sandwich (DAS)-ELISA with IYSV-specific serological reagents from Agdia Inc. (Elkhart, IN). A reverse transcription (RT)-PCR assay was used to verify the presence of IYSV in a subset of symptomatic leaf samples that reacted to IYSV antibodies in DAS-ELISA. Primers specific to the nucleocapsid (N) gene of IYSV (5′-ACTCACCAATGTCTTCAAC-3′ and 5′-GGCTTCCTCTGGTAAGTGC-3′) were used to characterize a 402-bp fragment (3). The resulting amplicons were ligated in TOPO TA cloning vector (Invitrogen, Carlsbad, CA) and two clones of each isolate were sequenced in both directions. Sequence analysis showed a consensus sequence for the partial N gene of the five IYSV isolates from Pennsylvania (GenBank Accession No. JQ952568) and an 87 to 100% nucleotide sequence identity with other IYSV N gene sequences that are available in GenBank. The highest nucleotide sequence identity (100%) was with an IYSV isolate from Texas (GenBank Accession No. DQ658242) and the lowest was with an isolate from India (GenBank Accession No. EU310291). To our knowledge, this is the first report of IYSV infection of onion in Pennsylvania. This finding confirms further spread of the virus within North America. Further study is warranted to determine the impact of IYSV on the Pennsylvania onion industry and to determine viable management strategies, if necessary. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004 (2) C. A. Hoepting et al. Plant Dis. 91:327, 2007 (3) C. L. Hsu et al. Plant Dis. 95:735-743. (4) S. W. Mullis et al. Plant Dis. 88: 1285, 2004.

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First Report of Iris yellow spot virus on Onion in Canada

Plant Disease ◽

10.1094/pdis-92-2-0318a ◽

2008 ◽

Vol 92 (2) ◽

pp. 318-318 ◽

Cited By ~ 8

Author(s):

C. A. Hoepting ◽

J. K. Allen ◽

K. D. Vanderkooi ◽

M. Y. Hovius ◽

M. F. Fuchs ◽

...

Keyword(s):

United States ◽

Nucleotide Sequence ◽

Nucleotide Sequence Identity ◽

Yellow Spot ◽

Home Garden ◽

N Gene ◽

Spot Virus ◽

Sequence Identity ◽

Symptomatic Tissue ◽

Das Elisa

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of onion vectored by onion thrips (Thrips tabaci Lindeman). Rapid spread of IYSV has occurred in the western United States and Georgia, with recent reports of IYSV from New York in the northeastern United States (1). In June and mid-July of 2007, symptomatic plants were found in Ontario, Canada in onions grown from sets in a home garden in Grey County (44°27′N, 80°53′W) and on a small commercial farm in Ottawa-Carleton County (45°14′N, 75°28′W), respectively. In the home garden, bleached, elongated lesions with tapered ends occurred on middle-aged leaves of 30% of 100 plants. By August 2007, 91% of these plants were symptomatic. In Ottawa-Carleton, two lesions with green centers and yellow borders occurred on a single leaf of a single plant in a field of 1,120 plants. Symptomatic leaf tissue tested positive for IYSV by IYSV-specific antiserum from Agdia Inc. (Elkhart, IN) in a double-antibody sandwich (DAS)-ELISA. These two isolated and remote finds of IYSV in Ontario prompted a survey in early August of 2007 of the Holland Marsh (44°5′N, 79°35′W), the largest onion-producing region in Ontario. Nine onion fields separated geographically across the Holland Marsh Region were scouted and one to three samples of symptomatic tissue per field were analyzed by DAS-ELISA. IYSV was confirmed in seven of nine (78%) fields surveyed and in 13 of 16 (81%) of the individual samples. A reverse transcription (RT)-PCR assay was used to verify the presence of IYSV in one new symptomatic tissue sample per field collected from three of the fields where IYSV was confirmed by ELISA. Primers specific to the small (S) RNA of IYSV (5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′) were used (2). The resulting 1.2-kb amplicon, which included the 772-bp nucleocapsid (N) gene was cloned and sequenced. Sequence analysis showed that the N gene of the Ontario isolate (GenBank Accession No. EU287943) shared 92 to 98% nucleotide sequence identity with known IYSV N gene sequences. The highest nucleotide sequence identity (98%) was with Genbank Accession Nos. DQ233475 and DQ233472. To our knowledge, this is the first report of IYSV infection of onion in Ontario and Canada. This finding 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 production. The finding of IYSV in remote and isolated locations where onions were grown from sets implies that the spread of IYSV via infected bulbs warrants further investigation as a potentially important route of distribution of the virus. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006.

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First Report of Iris yellow spot virus Infecting Onion in Kenya and Uganda

Plant Disease ◽

10.1094/pdis-01-11-0057 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1195-1195 ◽

Cited By ~ 7

Author(s):

R. Birithia ◽

S. Subramanian ◽

H. R. Pappu ◽

P. Sseruwagi ◽

J. W. Muthomi ◽

...

Keyword(s):

Sri Lanka ◽

Nucleotide Sequence ◽

Nucleotide Sequence Identity ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Spot Virus ◽

Viral Pathogen ◽

First Report ◽

Sequence Identity ◽

Small Holder

Onion (Allium cepa L.) is one of the key vegetables produced by small-holder farmers for the domestic markets in Sub-Saharan Africa. Biotic factors, including infestation by thrips pests such as Thrips tabaci Lindeman, can inflict as much as 60% yield loss. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) transmitted by T. tabaci is an economically important viral pathogen of bulb and seed onion crops in many onion-growing areas of the world (2,4). In Africa, IYSV has been reported in Reunion (1) and South Africa (3). In September 2009, symptoms suspected to be caused by IYSV were observed on onions and leeks cultivated in Nairobi, Kenya. Symptoms consisted of spindle-shaped, straw-colored, irregular chlorotic lesions with occasional green islands on the leaves. The presence of the virus was confirmed with IYSV-specific Agdia Flash kits (Agdia Inc., Elkart, IN). Subsequently, surveys were undertaken in small-holder farms in onion production areas of Makueni (January 2010) and Mwea (August 2010) in Kenya and Kasese (January 2010) and Rwimi (January 2010) in Uganda. The incidence of disease in these locations ranged between 27 and 72%. Onion leaves showing symptoms of IYSV infection collected from both locations tested positive for the virus by double-antibody sandwich-ELISA with IYSV-specific antiserum (Agdia Inc). IYSV infection was confirmed by reverse transcription-PCR with primers IYSV-465c: 5′-AGCAAAGTGAGAGGACCACC-3′ and IYSV-239f: 5′-TGAGCCCCAATCAAGACG3′ (3) as forward and reverse primers, respectively. Amplicons of approximately 240 bp were obtained from all symptomatic test samples but not from healthy and water controls. The amplicons were cloned and sequenced from each of the sampled regions. Consensus sequence for each isolate was derived from at least three clones. The IYSV-Kenya isolate (GenBank Accession No. HQ711616) had the highest nucleotide sequence identity of 97% with the corresponding region of IYSV isolates from Sri Lanka (GenBank Accession No. GU901211), followed by the isolates from India (GenBank Accession Nos. EU310287 and EU310290). The IYSV-Uganda isolate (GenBank Accession No. HQ711615) showed the highest nucleotide sequence identity of 95% with the corresponding region of IYSV isolates from Sri Lanka (GenBank Accession No. GU901211) and India (95% with GenBank Accession Nos. EU310274 and EU310297). To our knowledge, this is the first report of IYSV infecting onion in Kenya and Uganda. Further surveys and monitoring of IYSV incidence and distribution in the region, along with its impact on the yield, are under investigation. References: (1) L. J. du Toit et al. Plant Dis. 91:1203, 2007. (2) D. H. Gent et al. Plant Dis. 88:446, 2004. (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 Iris yellow spot virus in Onion in Mauritius

Plant Disease ◽

10.1094/pdis-10-09-0645 ◽

2010 ◽

Vol 94 (11) ◽

pp. 1373-1373 ◽

Cited By ~ 5

Author(s):

K. Lobin ◽

A. Saison ◽

B. Hostachy ◽

S. P. Benimadhu ◽

H. R. Pappu

Keyword(s):

Disease Incidence ◽

Consensus Sequence ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Spot Virus ◽

Viral Pathogen ◽

First Report ◽

Das Elisa

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) transmitted by thrips (Thrips tabaci Lindeman) is an economically important viral pathogen of bulb and seed onion (Allium cepa) crops in many onion-growing areas of the world (2,3). In Africa, IYSV has been reported in Reunion (4) and South Africa (1). In June 2008, diamond-shaped lesions that are typical of IYSV were observed on onion seed scapes in an onion plot of 0.25 ha at Reduit in the central part of Mauritius. Disease incidence was 80% with a severity of 50 to 75% of the scape surface area. Lodging was observed in 25% of the symptomatic plants. Twenty-two symptomatic plants were tested and found to be positive for IYSV when tested by double antibody sandwich (DAS)-ELISA with a commercially available kit (Agdia Inc., Elkhart, IN). The presence of the virus was confirmed by reverse transcription (RT)-PCR tests with primers 917L: 5′-TAAAACTTAACTAACACAAA-3′ and 56U: 5′-TCCTAAGTATTCACCAT-3′ as forward and reverse primers, respectively, for specific sequences flanking the CP gene. Another set of primers specific to the small (S) RNA of IYSV (5′-TAAAACAAACATTCAAACAA-3′ and 5′-CTCTTAAACACATTT AACAAGCAC-3′) produced an amplicon of approximately 1.2 kb that includes the 772-bp nucleocapsid (N) gene. The 1.2-kb amplicon was cloned and four clones were sequenced and consensus sequence was used for comparisons. Sequence analysis showed that the N gene of the IYSV isolate from Mauritius (GenBank Accession No. HM218822) shared the highest nucleotide sequence identity (99%) with several known IYSV N gene sequences (Accession Nos. FJ785835 and AM900393) available in the GenBank, confirming the presence of IYSV in the onion crops in Mauritius. A survey was subsequently carried out from July to November 2008 in major onion-growing localities at La Marie, Henrietta, Reduit, and Plaine Sophie (center); Bassin, La Ferme, and La Chaumiere (west); Grand Sable, Petit Sable, and Plaisance (south, southeast); and Belle Mare, Trou d'Eau Douce, and Palmar (east) to monitor the distribution of the disease on the island. Symptomatic samples with diamond-to-irregularly shaped lesions were observed and 155 symptomatic and 35 nonsymptomatic samples were collected and screened by DAS-ELISA for IYSV and Tomato spotted wilt virus (TSWV), another tospovirus reported to infect onion elsewhere. Sixty-six percent of the symptomatic samples screened (102 of 155) tested positive for IYSV. No IYSV was detected in the symptomless samples. There was no serological indication of TSWV infection in the samples. Samples that tested positive for IYSV were collected from Belle mare, Palmar, and Trou d'eau douce in the east and La Ferme in the west. Cultivars infected were Gandiole, Local Red, and Veronique. No IYSV was detected in the bulbs. The vector, T. tabaci, was observed in infected onion parcels surveyed and is known to occur in all onion-producing areas of the island. To our knowledge, this is the first report of IYSV in onion in Mauritius. Further surveys and monitoring of IYSV incidence, along with its impact on the yield, need to be established. References: (1) L. J. du Toit et al. Plant Dis. 91:1203, 2007. (2) D. H. Gent et al. Plant Dis. 88:446, 2004. (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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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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Green Foxtail (Setaria viridis), A Naturally Infected Grass Host of Iris yellow spot virus in Utah

Plant Disease ◽

10.1094/pdis-93-6-0670c ◽

2009 ◽

Vol 93 (6) ◽

pp. 670-670 ◽

Cited By ~ 11

Author(s):

C. K. Evans ◽

S. Bag ◽

E. Frank ◽

J. Reeve ◽

C. Ransom ◽

...

Keyword(s):

Nucleotide Sequence ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Grass Species ◽

Setaria Viridis ◽

Onion Thrips ◽

Spot Virus ◽

First Report ◽

Green Foxtail

Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is a serious virus pathogen in onion bulb and seed crops in the United States and several parts of the world (1). The virus is exclusively transmitted by onion thrips (Thrips tabaci). Besides onion and other susceptible crops such as garlic, leek, chives, and several ornamentals, weeds could be serving as potential reservoir sources of virus inoculum. There are reports of several weeds found naturally infected with IYSV (1,2,4). However, there is no report of IYSV infection of a grass species. Leaves of green foxtail (Setaria viridis (L.) Beauv.) were collected from two naturally occurring plants approximately 30 m apart in a weed trial conducted in commercial onions grown in Box Elder County, UT on 24 September 2008. Notes of IYSV symptoms on green foxtail were made only on the two grass plants sampled. Density of green foxtail in the weed trial was low and was not recorded. Leaves on both plants displayed a range of symptoms that included streaking, purpling, and chlorotic and necrotic lesions along leaf margins oriented along the axis of longitudinal venation. Samples were positive for IYSV by double-antibody sandwich-ELISA with a commercially available kit (Agdia Inc., Elkhart, IN). ELISA values of the grass samples were 2.64 and 2.23 for each plant sampled. Negative and positive control readings were 0.24 and 4.33, respectively. All absorbance readings were made at 405 nm. To provide a contrast of the grass data in context to the onion field where the weed trial was located, final visual assessments of onions in the field were made on 4 September 2009. Approximately 300 onion plants were assessed for incidence and severity of disease. Incidence of the disease among onions was 100% and the severity of iris yellow spot on leaves was 20 lesions per leaf. The average ELISA value over 30 individual onions arbitrarily sampled from the field on the same day was 3.50, and the ELISA values among the samples ranged from 1.37 to 4.38. The negative and positive controls were 0.19 and 4.40, respectively. To further verify the presence of IYSV in the grass specimen, reverse transcription-PCR was performed on total nucleic acid extracts obtained from the symptomatic parts of the leaves. Primers specific to the nucleocapsid (N) gene coded by the small (S)-RNA of IYSV were used (3). The forward and reverse primer pairs, 5′-TCAGAAATCGAGAAACTT-3′ and 5′-CACCAATGTCTTCAACAATCTT-3′, respectively, amplify a 751-nt fragment of the N gene (3). An amplicon of expected size was obtained, cloned, and sequenced. The nucleotide sequence analysis and comparison with known IYSV S-RNA sequences showed that the amplicon from foxtail (GenBank Accession No. FJ652594) samples had the highest nucleotide sequence identity (98%) with the corresponding region of an IYSV isolate from Jefferson County, OR (GenBank Accession No. DQ233479). To our knowledge, this is the first report of natural infection of a grass species by IYSV and the first report of a Tospovirus infecting a grass species. The data suggests grasses may serve as a new host reservoir for IYSV. The increasing number of weed hosts of IYSV warrants further study on the role of these weeds as hosts for onion thrips and in IYSV epidemiology. References: (1) D. Gent et al. Plant Dis. 90:1468, 2006. (2) C. Nischwitz et al. Plant Dis. 91:1518, 2007. (3) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (4) R. Sampangi et al. Plant Dis. 91:1683, 2007.

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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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First Report of Onion (Allium cepa) Naturally Infected with Iris yellow spot virus in Peru

Plant Disease ◽

10.1094/pd-90-0377a ◽

2006 ◽

Vol 90 (3) ◽

pp. 377-377 ◽

Cited By ~ 8

Author(s):

S. W. Mullis ◽

R. D. Gitaitis ◽

C. Nischwitz ◽

A. S. Csinos ◽

Z. C. Rafael Mallaupoma ◽

...

Keyword(s):

The United States ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Enzyme Linked Immunosorbent Assay ◽

Spot Virus ◽

First Report ◽

The Impact ◽

Das Elisa

Onions have become an important export crop for Peru during the last few years. The onions produced for export are primarily short-day onions and include Grano- or Granex-type sweet onions. The first of two growing seasons for onion in Peru occurs from February/March until September/October and the second occurs from September/October to December/January. Iris yellow spot virus (IYSV [family Bunyaviridae, genus Tospovirus]), primarily transmitted by onion thrips (Thrips tabaci), has been reported in many countries during recent years, including the United States (1,2). In South America, the virus was reported in Brazil during 1999 (3) and most recently in Chile during 2005 (4). During 2003, an investigation of necrotic lesions and dieback in onions grown near the towns of Supe and Ica, Peru led to the discovery of IYSV in this region. Of 25 samples of symptomatic plants collected from five different fields near Supe, 19 tested strongly positive and an additional three tested weakly positive for IYSV using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) (Agdia Inc., Elkhart, IN). None of the samples tested positive for Tomato spotted wilt virus (TSWV). A number of onions with necrosis and dieback symptoms were also observed during 2004 and 2005. During September 2005, 25 plants with symptoms suspected to be caused by IYSV or TSWV in the Supe and Casma valleys were collected and screened for both viruses using DAS-ELISA. All plants screened were positive for IYSV. There was no serological indication of TSWV infection in these samples. The positive samples were blotted onto FTA cards (Whatman Inc., U.K.) to bind the viral RNA for preservation and processed according to the manufacturer's protocols. The presence of IYSV was verified by reverse transcription-polymerase chain reaction (RTPCR) using (5′-TCAGAAATCGAGAAACTT-3′) and (5′-TAATTATATCTATCTTTCTTGG-3′) as forward and reverse primers (1), respectively. The primers amplify the nucleocapsid (N) gene of IYSV, and the RT-PCR products from this reaction were analyzed with gel electrophoresis with an ethidium bromide stain in 0.8% agarose to verify the presence of this amplicon in the samples. Subsequent to the September 2005 sampling, 72 additional samples from regions in northern and southern Peru were analyzed in the same manner. The amplicons obtained were cloned, sequenced, and compared with known IYSV isolates for further verification. Onions have become a significant export crop for Peru, and more research is needed to determine the impact of IYSV on the Peruvian onion export crop. To our knowledge, this is the first report of IYSV in onion in Peru. References: (1) L. du Toit et al. Plant Dis. 88:222, 2004. (2) S. W. Mullis et al. Plant Dis. 88:1285, 2004. (3) L. Pozzer et al. Plant Dis. 83:345, 1999. (4) M. Rosales et al. Plant Dis. 89:1245, 2005.

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First Report of Iris yellow spot virus Infection of Garlic and Egyptian Leek in Egypt

Plant Disease ◽

10.1094/pdis-10-11-0902 ◽

2012 ◽

Vol 96 (4) ◽

pp. 594-594 ◽

Cited By ~ 6

Author(s):

E. E. Hafez ◽

A. A. Abdelkhalek ◽

A. A. El-Morsi ◽

O. A. El-Shahaby

Keyword(s):

Plant Species ◽

Natural Infection ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Sri Lankan ◽

Spot Virus ◽

Viral Pathogen ◽

First Report ◽

Sequence Identity ◽

Das Elisa

Egyptian leek (Allium ampeloprasum), garlic (A. sativum), and onion (A. cepa) are key vegetables produced by small- and large-scale farmers in Egypt for national and international markets. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of bulb and seed onion crops in many onion-growing areas of the world (1,3). During February and March of 2011, symptoms of spindle-shaped, straw-colored, irregular lesions with occasional green islands were observed on onion, garlic, and Egyptian leek cultivated on large and small farms in Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiut governorates in Egypt. The presence of IYSV was confirmed by specific double antibody sandwich (DAS)-ELISA Flash Kits (Agdia Inc., Elkhart, IN) (2). A survey was carried out by collecting 100 plant samples (10 asymptomatic and 90 symptomatic) of each plant species from fields in the governorates of Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiute and testing the plants using DAS-ELISA. For onion and garlic, 45% of the symptomatic samples and 0% of the asymptomatic plants tested positive. For leek, 34% of the symptomatic samples tested positive and 0% of the asymptomatic samples. ELISA-positive samples were tested using a reverse transcription (RT)-PCR assay with primers specific to the S RNA of IYSV (forward primer 5′-TAAAACAAACATTCAAACAA-3′ and reverse primer 5′-CTCTTAAACACATTTAACAAGCAC-3′) (2). Amplicons of approximately 1,100 bp were obtained from all symptomatic samples that were ELISA positive, but none of the asymptomatic plants nor the sterile water control sample produced PCR amplicons. The amplicons were cloned (at least three clones per plant species) using the TOPO TA Cloning Kit (Invitrogen, Grand Island, NY), and sequenced. The Egyptian onion IYSV isolate (GenBank No. JN541273) had the greatest nucleotide sequence identity (86%) with the corresponding S RNA region of IYSV isolates from India (GenBank Nos. EU310290, EU310284, and EU310276). The Egyptian garlic IYSV isolate (GenBank No. JN541275) showed the strongest identity (93%) with that of a Sri Lankan IYSV isolate (GenBank No. GU901211). The Egyptian leek IYSV isolate (GenBank No. JN541274) exhibited 91% sequence identity with that of the same Sri Lankan isolate (No. GU901211). To our knowledge, this is the first report of IYSV infecting garlic and Egyptian leek in Egypt. IYSV infection of onion was reported previously from the agricultural farm of the Faculty of Agriculture, Cairo University, Giza (4), but to our knowledge, this is the first report of natural infection by the virus in commercial onion production in Egypt. Further surveys and monitoring of IYSV incidence and distribution in the entire Egyptian governorate are under investigation. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) A. Manal et al. Egypt. J. Virol. 3:49, 2006.

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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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