Detección de Iris yellow spot virus en cebolla cultivada en Tepalcingo, Morelos, México

En el estado de Morelos recientemente se han observado enfermedades virales en cebolla (Allium cepa); una de ellas es la mancha amarilla causada por el Iris yellow spot virus perteneciente a la familia Bunyaviridae del género Tospovirus, el cual se trasmite a la cebolla por Thrips tabaci Lindeman (Thysanoptera: Thripidae). En el 2012 la incidencia de esta enfermedad fue de 100 % en las 2,500 ha cultivadas en la entidad con una severidad superior a 90 %. El objetivo de este trabajo fue detectar mediante RT-PCR en tiempo real y secuenciar la presencia de IYSV. Para su identificación se tomaron muestras de hojas de cebolla con manchas amarillentas alargadas, desde el trasplante hasta la cosecha en Tepalcingo, Morelos. La extracción de ARN total se realizó utilizando TRIzol® Reagent. La detección del virus se realizó con primers específicos al gen de la nucleoproteína de IYSV dando como resultado la amplificación de un producto específico mediante RT-PCR en tiempo real y una banda esperada de 896 pb la cual mediante secuenciación confirmó 99 % de identidad con el gen de la nucleoproteína de este virus. El IYSV fue detectado en plantas de cebolla en Tepalcingo, Morelos y su secuencia se registró en la base de datos del GenBank (JX946658).

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Hospedantes alternos de Iris yellow spot virus y trips en cultivos de cebolla de Morelos y Michoacán, México

Revista Mexicana de Fitopatología Mexican Journal of Phytopathology ◽

10.18781/r.mex.fit.1701-1 ◽

2017 ◽

Vol 35 (2) ◽

Author(s):

Norma Ávila Alistac ◽

Sergio Ramírez Rojas ◽

Ángel Rebollar Alviter ◽

Remigio Anastacio Guzmán Plazola

Keyword(s):

Allium Cepa ◽

Ricinus Communis ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Rt Pcr ◽

Spot Virus ◽

Das Elisa

El objetivo de la investigación fue identificar hospedantes alternos de Iris yellow spot virus (IYSV) y establecer el rango de hospedantes del putativo vector(es) en regiones productoras de cebolla (Allium cepa) de Morelos y Michoacán, México. En 10 localidades de ambos estados se muestrearon cultivos de cebolla y arvenses, en presencia y ausencia del cultivo. Las plantas de cebolla se analizaron por RT-PCR y las arvenses por DAS-ELISA. Las arvenses se identificaron a nivel especie, los trips colectados de las mismas se establecieron colonias para su identificación por PCR, con iniciadores específicos que amplifican un segmento del gen de citocromo oxidasa I (COI). Se analizaron e identificaron 295 arvenses agrupadas en 56 especies (23 familias), todas resultaron negativas para IYSV. Se detectaron trips en 75 arvenses agrupadas en 17 especies. Se analizaron 33 poblaciones de trips (22 de Morelos y 11 de Michoacán). La secuenciación indicó identidad con Thrips tabaci con una homología superior a 97 %. Las arvenses Ricinus communis y Acalypha ostryifolia registraron el mayor número de T. tabaci. En cebolla se confirmó la presencia de IYSV con RT-PCR en las 10 parcelas muestreadas. Este es el primer reporte de la presencia de IYSV en el estado de Michoacán.

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Hospedantes alternos de Iris yellow spot virus y trips en cultivos de cebolla de Morelos y Michoacán, México

Revista Mexicana de Fitopatología Mexican Journal of Phytopathology ◽

10.18781/10.18781/r.mex.fit.1701-1 ◽

2017 ◽

Vol 35 (2) ◽

Author(s):

Norma Ávila Alistac ◽

Sergio Ramírez Rojas ◽

Ángel Rebollar Alviter ◽

Remigio Anastacio Guzmán Plazola

Keyword(s):

Allium Cepa ◽

Ricinus Communis ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Rt Pcr ◽

Spot Virus ◽

Das Elisa

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Potential factors contributing to the decline of Iris Yellow Spot Virus in organically grown onion crops in Canterbury and Hawke's Bay over three years

New Zealand Plant Protection ◽

10.30843/nzpp.2018.71.116 ◽

2018 ◽

Vol 71 ◽

pp. 39-44

Author(s):

Melanie M. Davidson ◽

Mette C. Nielson ◽

John D. Fletcher

Keyword(s):

New Zealand ◽

Allium Cepa ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Spot Virus ◽

Potential Factors ◽

Organically Grown ◽

Organically Managed

The Tospovirus Iris yellow spot virus (IYSV), transmitted by thrips (predominantly Thrips tabaci), was first recorded in New Zealand in 2007. In March 2015, symptoms of the virus were relatively widespread in an organically managed onion crop in Canterbury. Onion plants were sampled for the presence of T. tabaci adults and larvae and for IYSV symptoms on an organically managed farm in Canterbury in 2014-2015, 2015-16 and 2016-17, and on a similar farm in Hawke's Bay in 2014-2015 and 2015-16. An immunoassay was used to confirm the presence of IYSV in some symptomatic plants. In Canterbury, IYSV symptoms were less apparent in 2015-16 and no symptoms were observed in 2016-17. No IYSV symptoms were observed in the Hawke's Bay onion crop, despite relatively high T. tabaci numbers. The virus symptoms declined from when they were first observed in March 2015 to undetectable levels in 2016-17 in Canterbury, which may be attributed to crop location, fewer thrips and the absence of a disease reservoir in volunteer Allium cepa plants or other hosts.

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

Plant Disease ◽

10.1094/pdis.2004.88.11.1285b ◽

2004 ◽

Vol 88 (11) ◽

pp. 1285-1285 ◽

Cited By ~ 19

Author(s):

S. W. Mullis ◽

D. B. Langston ◽

R. D. Gitaitis ◽

J. L. Sherwood ◽

A. C. Csinos ◽

...

Keyword(s):

Tomato Spotted Wilt Virus ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Unknown Etiology ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Spot Virus ◽

Tomato Spotted Wilt ◽

Wilt Virus

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

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DETECCIÓN DEL IRIS YELLOW SPOT VIRUS EN EL CULTIVO DE CEBOLLA EN ZACATECAS, MÉXICO

Revista Mexicana de Ciencias Agrícolas ◽

10.29312/remexca.v2i6.1598 ◽

2018 ◽

Vol 2 (6) ◽

pp. 971-978

Author(s):

Rodolfo Velásquez-Valle ◽

Manuel Reveles-Hernández

Keyword(s):

Frankliniella Occidentalis ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Spot Virus ◽

La Mancha ◽

Das Elisa

El virus de la mancha amarilla del iris (IYSV) es la enfermedad viral de mayor importancia para los cultivos de ajo y cebolla en Zacatecas, México. A finales de mayo de 2010 se encontraron lesiones amarillentas en forma de diamante en hojas y tallo floral de plantas de cebolla en parcelas comerciales, localizados en los municipios de Loreto, Villa de Cos, Sain Alto, Villanueva, Calera de V. R. y Enrique Estrada pertenecientes al estado de Zacatecas. La prueba DAS-ELISA mostró resultados positivos para este virus en las muestras procedentes de diferentes municipios. Thrips tabaci el único vector conocido del IYSV se encontró en las parcelas comerciales muestreadas, aunque la presencia de Frankliniella occidentalis también fue reconocida en una parcela comercial de cebolla, cuyas muestras resultaron positivas a IYSV. No se encontró diferencia entre la altura, número de hojas, peso y diámetro de bulbo de plantas sin lesiones y aquellas con diferente número de lesiones de IYSV.

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Wild Allium spp. as Natural Hosts of Iris yellow spot virus

Plant Disease ◽

10.1094/pd-90-0378a ◽

2006 ◽

Vol 90 (3) ◽

pp. 378-378 ◽

Cited By ~ 7

Author(s):

H. R. Pappu ◽

B. C. Hellier ◽

F. M. Dugan

Keyword(s):

Natural Infection ◽

The State ◽

Thrips Tabaci ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

N Gene ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Spot Virus ◽

Sequence Comparisons

The incidence of Iris yellow spot virus (IYSV) of genus Tospovirus, family Bunyaviridae in a commercial onion crop was first confirmed in Washington state during 2003 (1). First found in Adams County, IYSV has rapidly spread to all onion-producing counties in the state, affecting seed and bulb crops. The USDA-ARS Western Regional Plant Introduction Station (WRPIS) collects, maintains, and distributes various Allium (garlic and onion) accessions. As part of the regeneration process, accessions are grown under field conditions at the WRPIS farms in two locations: Pullman and Central Ferry, WA. Symptoms indicative of viral infection, now known to be caused by IYSV, first appeared in field-grown accessions in 1999. In June 2005, leaf and scape tissues were collected from WRPIS accessions of wild onions (Allium pskemense, A. vavilovii, and A. altaicum) in Central Ferry that had symptoms indicative of IYSV infection (2). IYSV infection was confirmed using enzyme-linked immunosorbent assay with a commercially available kit (Agdia Inc., Elkhart, IN). Virus infection was further verified using reverse transcription-polymerase chain reaction (RT-PCR) with primers derived from the small (S) RNA of IYSV. The primers flanked the IYSV N gene (5′-TAA AAC AAA CAT TCA AAC AA-3′ and 5′-CTC TTA AAC ACA TTT AAC AAG CAC-3′). RT-PCR gave a PCR product of expected size (≈1.2 kb). The DNA amplicon was cloned and sequenced. Nucleotide sequence comparisons with known IYSV N gene sequences showed 95 to 98% sequence identity. The prevalence of the vector, onion thrips (Thrips tabaci), combined with the widespread incidence of IYSV in seed and bulb production areas of the state may have resulted in natural infection of wild relatives of cultivated onion. The potential role of wild Allium spp. in IYSV epidemiology remains to be determined. Information on the extent of IYSV infection of onion germplasm would be useful in identifying potential sources of host plant resistance to IYSV. References: (1) L. J. du Toit et al. Plant Dis. 88:222, 2004. (2) B. Hellier et al. APSnet Image of the Week. Online publication, iw000049.asp, 2004.

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

Plant Disease ◽

10.1094/pdis-92-8-1247a ◽

2008 ◽

Vol 92 (8) ◽

pp. 1247-1247 ◽

Cited By ~ 8

Author(s):

A. Bulajić ◽

J. Jović ◽

S. Krnjajić ◽

M. Petrov ◽

I. Djekić ◽

...

Keyword(s):

Allium Cepa ◽

Disease Incidence ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Mechanical Transmission ◽

Rt Pcr ◽

Spot Virus ◽

First Report ◽

Seed Crop ◽

Das Elisa

Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) is established in several European countries (France, Italy, The Netherlands, Poland, Slovenia, Spain, and the UK) and its distribution in the EU region has increased since 2002 (3). In July 2007, symptoms resembling those of IYSV were observed in an onion (Allium cepa) seed crop in the Sirig locality in Serbia. Onion plants exhibited characteristic symptoms of chlorotic or necrotic spindle and diamond-shaped lesions on the leaves and scapes. Symptomatic plants were found throughout the field and disease incidence was estimated at 80%. Leaf and scape samples were tested for the presence of IYSV and two other tospoviruses, Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV), using commercial double-antibody sandwich (DAS)-ELISA diagnostic kits (Loewe Biochemica, Sauerlach, Germany). All samples tested negative for TSWV and INSV. IYSV was detected serologically in 26 of 34 onion samples. To determine an experimental host range, samples of IYSV-infected onion plants were homogenized in chilled 0.05 M phosphate buffer pH 7 containing 1 mM Na-EDTA, 5 mM Na-DIECA, and 5 mM Na-thioglycolate (2), and host plants were inoculated with the sap. Mechanical transmission of the virus occurred rarely. All inoculated test plants were assayed by DAS-ELISA and only four species tested positive for IYSV, but not in all replications. Inoculated Chenopodium quinoa developed local chlorotic lesions, Nicotiana tabacum cvs. Samsun and Prilep showed mild mosaic, while infected N. benthamiana were symptomless. For further confirmation of IYSV, conventional reverse transcription (RT)-PCR was performed on extracts made from symptomatic onion leaf material and from the ELISA-positive symptomless leaves of N. benthamiana. Total RNAs were extracted with an RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was carried out with the OneStep RT PCR Kit (Qiagen) following the manufacturer's instructions. The primer pair, IYSV56U/IYSV917L, covering the entire nucleocapsid (NC) gene was used for both amplification and sequencing (1). A product of the correct predicted size (896 bp) was obtained from each of the plants assayed, and that derived from isolate 605-SRB was purified (QUIAqick PCR Purification Kit, Qiagen) and sequenced (GenBank Accession No. EU586203). BLAST analyses revealed 86 to 97% sequence identity with the NC gene from all other IYSV. The highest identity (97%) was with leek and onion isolates (GenBank Accession Nos. EF427447 and EF19888) from Spain. To our knowledge, this is the first report of IYSV infection of onion seed crop in Serbia. Thorough inspections and subsequent testing would be needed to establish the distribution and incidence of IYSV in Serbia. References: (1) I. Robène-Soustrade et al. Plant Pathol. 55:288, 2006. (2) P. Roggero et al. Plant Dis. 86:950, 2002. (3) C. Sansford and J. Woodhall. Pest Risk Analysis for Iris Yellow Spot Virus. Online publication. Central Science Laboratory, Sand Hutton, UK, 2007.

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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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Variabilidad y síntomas causados por Iris yellow spot virus en Nicotiana benthamiana

Revista Mexicana de Fitopatología Mexican Journal of Phytopathology ◽

10.18781/r.mex.fit.1707-2 ◽

2017 ◽

Vol 35 (4) ◽

Author(s):

Katya Ornelas-Ocampo ◽

Daniel Leobardo Ochoa-Martínez ◽

Sergio Aranda-Ocampo ◽

Sergio Ramírez-Rojas ◽

Hernán García-Ruíz

Keyword(s):

Allium Cepa ◽

Nicotiana Benthamiana ◽

Iris Yellow Spot Virus ◽

Yellow Spot ◽

Spot Virus

En cultivos de cebolla Allium cepa del estado de Morelos, México, se observan síntomas típicos y severos asociados a Iris yellow spot virus (IYSV). En esta investigación se estudiaron las alteraciones que ocasionan los aislamientos de IYSV procedentes de síntomas típicos y severos en Nicotiana benthamiana, las diferencias en el gen N y su filogenia. Cuatro aislamientos típicos y cinco severos inoculados mecánicamente causaron infección sistémica. En cámara bioclimática los aislamientos severos ocasionaron mayor severidad de síntomas. La secuencia del gen N de ambos aislamientos tuvo 98-99% de identidad con la nucleoproteína de IYSV y no se observaron cambios en la secuencia de nucleótidos entre ellos. Ambos aislamientos se agruparon con el genotipo IYSVBR y tuvieron mayor similitud con los reportados en Canadá y Estados Unidos.

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