The first report of Tomato brown rugose fruit virus in tomato in Norway

Plant Disease ◽

10.1094/pdis-10-21-2208-pdn ◽

2021 ◽

Author(s):

Zhibo Hamborg ◽

Dag-Ragnar Blystad

Keyword(s):

Uranyl Acetate ◽

Test Plant ◽

Surveillance Program ◽

Rt Pcr ◽

Potential Threat ◽

Greenhouse Production ◽

First Report ◽

Solanum Lycopersicum L ◽

One Step ◽

Rigid Rod

In May 2021, tomato (Solanum lycopersicum L.) plants with necrosis and ringspot symptoms were observed in a farm greenhouse at Sundbyfoss, Norway. This greenhouse production focused on ecological growing of several tomato varieties (i.e., “Blush Tiger”, “Sailor’s Luck”, “Evil Oliver”, etc.) with compost for local customers. The presence of tomato brown rugose fruit virus (ToBRFV) was suspected due to the symptoms in the diseased plants. Sap inoculation was carried out with extracts from 100 mg symptomatic tomato fresh leaves in 0.03 M PBS (phosphate buffered saline, pH 7.0), and inoculation on three Nicotiana tabacum cv. Xanthi plants at the 4-6 leaf stage grown in an experimental greenhouse. The test plants showed local necrotic lesions after 3-5 days. Two symptomatic leaf samples, one from tomato and the other from a test plant, were analysed by transmission electron microscope (TEM) with the help of negative staining. The samples were treated with 0.1 M PBS (pH 7.3) first and placed on carbon-coated EM grids and stained with 2% uranyl acetate before TEM observation. Rigid rod-shaped viral particles, typical of tobamovirus particls (around 300 nm) were observed. Total RNA was isolated from two tomato leaves showing symptoms using a Norgen Plant/Fungi RNA kit (Norgen Biotek, Canada). One-step reverse-transcription (RT)-PCR with specific primers ToBRFV-F/ToBRFV-R for ToBRFV (Alkowni et al. 2019), which amplified a 560-bp fragment, was performed. The sequence obtained by Sanger sequencing from the amplicon (535 nt) showed 99.8% nt identity with ToBRFV isolate PV-1241 (NCBI accession no. MZ202349) from DSMZ (Leibniz Institute, German) and was deposited in the GenBank database under the accession number OK358628. The infection was also confirmed with duplex real-time RT-PCR test for ToBRFV using CaTa28 and CSP1325 primers and probe (ISF 2020; EPPO, PM7/146 2021). In addition, eight tomato samples from the same farm greenhouse were collected according to cultivars, location in the greenhouse and symptoms before eradication and disinfection: four of the tomato samples were confirmed positive for ToBRFV with one-step RT-PCR as described above. A surveillance program for ToBRFV in commercial greenhouse production has been carried out in 2021 in Norway. Around 4000 tomato plants from 18 commercial tomato growers were tested. No positive ToBRFV samples have been detected. However, unregulated tomato seeds from abroad and self-propagation of tomato by private gardeners and hobby growers is a potential threat to commercial production of tomato in Norway. To our knowledge, this is the first report of ToBRFV associated with tomato in Norway and in the Nordic countries.

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First Report of Beet necrotic yellow vein virus Infecting Spinach in California

Plant Disease ◽

10.1094/pdis-94-5-0640a ◽

2010 ◽

Vol 94 (5) ◽

pp. 640-640 ◽

Cited By ~ 4

Author(s):

H.-Y. Liu ◽

B. Mou ◽

K. Richardson ◽

S. T. Koike

Keyword(s):

Spinacia Oleracea ◽

Sandwich Elisa ◽

Chenopodium Quinoa ◽

Yellow Vein ◽

Mechanical Transmission ◽

Polymyxa Betae ◽

Rt Pcr ◽

First Report ◽

Vein Clearing ◽

Rigid Rod

In 2009, plants from two spinach (Spinacia oleracea) experimental fields in Monterey County and one commercial spinach field in Ventura County of California exhibited vein-clearing, mottling, interveinal yellowing, and stunting symptoms. For experimental fields, up to 44% of spinach plants have symptoms. With a transmission electron microscope, rigid rod-shaped particles with central canals were observed from plant sap of the symptomatic spinach. Analysis with a double-antibody sandwich-ELISA assay for Beet necrotic yellow vein virus (BNYVV) showed that all 10 symptomatic plants we tested were positive and 5 asymptomatic plants were negative. Symptomatic spinach from both counties was used for mechanical transmission experiments. Chenopodium quinoa, Tetragonia expansa, and Beta vulgaris (sugar beet) showed chlorotic local lesions and B. macrocarpa and spinach showed vein-clearing, mottling, and systemic infections. To further confirm the presence of BNYVV, reverse transcription (RT)-PCR was conducted. Total RNA was extracted from field- and mechanically inoculated symptomatic spinach plants using an RNeasy Plant Kit (Qiagen Inc., Valencia, CA) and used as a template in RT-PCR. Forward and reverse primers specific to the BNYVV RNA-3 P25 protein gene from the beet isolate were used (2). Amplicons of the expected size (approximately 860 bp) were obtained. Four RT-PCR products were sequenced and the sequences were identical (GenBank Accession No. GU135626). Sequences from the spinach plants had 97 to 99% nucleotide and 94 to 100% amino acid identity with BNYVV RNA-3 P25 protein sequences available in the GenBank. On the basis of the data from electron microscopy, indicator plants, serology, and cDNA sequencing, the virus was identified as BNYVV. BNYVV has been reported from spinach fields in Italy (1). To our knowledge, this is the first report of BNYVV occurring naturally on spinach in California. Since BNYVV is transmitted by the zoospores of the soil-inhabiting plasmodiophorid Polymyxa betae, it could be a new threat to spinach production in the state. References: (1) C. R. Autonell et al. Inf. Fitopatol. 45:43, 1995. (2) H.-Y. Liu and R. T. Lewellen, Plant Dis. 91:847, 2007.

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First Report of Tobacco rattle virus in Spinach in California

Plant Disease ◽

10.1094/pdis-94-1-0125b ◽

2010 ◽

Vol 94 (1) ◽

pp. 125-125 ◽

Cited By ~ 3

Author(s):

S. T. Koike ◽

T. Tian ◽

H.-Y. Liu

Keyword(s):

Sugar Beet ◽

Spinacia Oleracea ◽

Sandwich Elisa ◽

Tobacco Rattle Virus ◽

Chenopodium Quinoa ◽

Mechanical Transmission ◽

Rt Pcr ◽

First Report ◽

Local Lesions ◽

Rigid Rod

In 2009 in coastal California (Santa Barbara County), commercially grown spinach (Spinacia oleracea) in two nearby fields exhibited symptoms of a previously unrecognized virus-like disease. Symptoms consisted of general chlorosis and bright yellow blotches and spots. Necrotic spots were also associated with the disease. In affected fields, disease occurred in limited, irregularly shaped patches that ranged from one to several meters in diameter. Symptomatic plants were unmarketable and these small patches of spinach were not harvested. With a transmission electron microscope, rigid, rod-shaped particles with a clear central canal were observed from plant sap of the symptomatic spinach. Analysis by a double-antibody sandwich-ELISA assay (Agdia Inc., Elkhart, IN) for Tobacco rattle virus (TRV) showed that the symptomatic plants were positive. Symptomatic spinach from the field was used for mechanical transmission to Chenopodium quinoa, C. murale, C. capitatum, spinach, and sugar beet (Beta vulgaris). All inoculated plants showed chlorotic local lesions and sugar beet showed chlorotic local lesions with rings. To further confirm the presence of TRV, reverse transcription (RT)-PCR was conducted. Total RNA was extracted from the mechanically inoculated symptomatic spinach plants using an RNeasy Plant Kit (Qiagen Inc., Valencia, CA) and used as a template in RT-PCR with forward (5′-TACATCACATCTGCCTGC-3′) and reverse (5′-CTTCATTCACACAACCCTTG-3′) primers specific to the movement protein gene from the spinach isolate of TRV (GenBank Accession No. AJ007294). Amplicons of the expected size (approximately 562 bp) were obtained. The RT-PCR products were sequenced (GenBank Accession No. GU002156) and compared with TRV sequences in GenBank to confirm the identity of the products. Sequences obtained had 96% nucleotide identity and 97% amino acid identity with TRV sequences available under the GenBank Accession Nos. FJ357571 and AJ007294. On the basis of the data from electron microscopy and serological and molecular analyses, the virus was identified as TRV. Soil samples collected from one of the fields were assayed for nematodes; however, Paratrichodorus or Trichodorus species were not recovered. To our knowledge, this is the first report of TRV in spinach in California. TRV has also been reported in spinach in England (1) and Germany (2). References: (1) A. Kurppa et al. Ann. Appl. Biol. 98:243, 1981. (2) K. Schmidt and R. Koenig. Arch. Virol. 144:503, 1999.

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First Report of Ranunculus mild mosaic virus on Ranunculus asiaticus in Yunnan Province, China

Plant Disease ◽

10.1094/pdis-92-11-1585a ◽

2008 ◽

Vol 92 (11) ◽

pp. 1585-1585 ◽

Cited By ~ 1

Author(s):

J. H. Wang ◽

S. Zhao ◽

X. M. Yang

Keyword(s):

Mosaic Virus ◽

Yunnan Province ◽

Disease Incidence ◽

Viral Disease ◽

Mild Mosaic ◽

Rt Pcr ◽

Potential Threat ◽

First Report ◽

Ranunculus Asiaticus ◽

Mild Mosaic Virus

In June 2007, a new viral disease occurred in commercial fields of Ranunculus asiaticus in the Yunnan Province of China. Infected plants exhibited mosaic symptoms and growth abnormalities. Viral disease incidence for this ornamental crop host in the Yunnan Province was estimated to range from 10 to 20%. Electron microscopic examination of negatively stained leaf-dip preparations from symptomatic plants identified long, flexuous linear particles (approximately 800 nm). The samples were tested using indirect antigen-coated plate (ACP)-ELISA. ACP-ELISA results showed that the leaf samples from symptomatic plants reacted positively to the potyvirus group antibody (Agdia Inc., Eklhart, IN). Total nucleic acid extracted from symptomatic plants was tested using reverse transcription (RT)-PCR with primers (S 5′-GGNAAAAYAGYGGNCARCC-3′; M4: 5′-GTTTTCCCAGTCACGAC-3′ [N = A, G, C, or T; Y = C or T; and R = A or G]) designed to amplify the 3′ terminal region of genomic RNA of the genus Potyvirus (1). RT-PCR produced a 1,650-bp amplification product that was cloned and sequenced (GenBank Accession No. EU684747). The sequenced portion showed 90 and 99% identity with the Ranunculus mild mosaic virus (RMMV) isolates (GenBank Accession Nos. DQ152191 and EF445546) from Italy and Israel, respectively (2). To our knowledge, this is the first report of RMMV in China. Infection from this virus may cause losses for cut-flower production of Ranunculus asiaticu and it is also a potential threat for international trade of Ranunculus germplasm. References: (1) J. Chen and J. P. Chen. Chin. J. Virol. 18:371, 2002. (2) M. Turina et al. Phytopathology 96:560, 2006.

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First Report of Cucumber green mottle mosaic virus on Melon in the United States

Plant Disease ◽

10.1094/pdis-02-14-0176-pdn ◽

2014 ◽

Vol 98 (8) ◽

pp. 1163-1163 ◽

Cited By ~ 35

Author(s):

T. Tian ◽

K. Posis ◽

C. J. Maroon-Lango ◽

V. Mavrodieva ◽

S. Haymes ◽

...

Keyword(s):

United States ◽

Mosaic Virus ◽

Seed Production ◽

Citrullus Lanatus ◽

The United States ◽

Rt Pcr ◽

First Report ◽

Rigid Rod ◽

Yolo County ◽

Das Elisa

In July 2013, a melon (Cucumis melo var. Saski) field in Yolo County, California, was inspected as part of a phytosanitary inspection for seed production. The leaves of the plants showed mosaic, green mottle, and blotches. When plant sap was examined using a transmission electron microscope, rigid rod-shaped particles were observed. Melon plant samples were analyzed by both CDFA and USDA APHIS PPQ laboratories and tested positive using DAS-ELISA against Cucumber green mottle mosaic virus (CGMMV) (Agdia, Elkhart, IN). To confirm the presence of CGMMV, total RNA was analyzed by RT-PCR using primers CGMMV-F5370 5′-CTAATTATTCTGTCGTGGCTGCGGATGC-3′ and CGMMV-R6390 5′-CTTGCAGAATTACTGCCCATA-3′ designed by PPQ based on 21 genomic sequences of CGMMV found worldwide. The 976-bp amplicon was sequenced (GenBank Accession No. KJ453559) and BLAST analysis showed the sequence was 95% identical to MP and CP region of CGMMV isolates reported from Russia (GQ495274, FJ848666), Spain (GQ411361), and Israel (KF155231), and 92% to the isolates from China (KC852074), Korea (AF417243), India (DQ767631), and Japan (D12505). These analyses confirm the virus was CGMMV. To our knowledge, this is the first report of CGMMV in the United States. Based on our sequence data, a second set of primers (CGMMV-F5796 5′-TTGCGTTTAGTGCTTCTTATGT-3′ and CGMMV-R6237 5′-GAGGTGGTAGCCTCTGACCAGA-3′), which amplified a 440-bp amplicon from CGMMV CP region, was designed and used for testing all the subsequent field and seed samples. Thirty-seven out of 40 randomly collected Saski melon samples tested positive for CGMMV, suggesting the virus was widespread in the field. All the melon samples also tested positive for Squash mosaic virus (SqMV) using DAS-ELISA (Agdia). Therefore, the symptoms observed likely resulted from a mixed infection. The melon field affected by CGMMV was immediately adjacent to fields of cucumber (Cucumis sativus var. Marketmore 76) and watermelon (Citrullus lanatus var. Sugar Baby) crops, both for seed production with no barrier between the crops. CGMMV was also detected from symptomatic plants from both fields. Seed lots used for planting all three crops were tested and only the melon seed was positive for CGMMV, suggesting the seed as the source of infection. The sequenced 440-bp RT-PCR amplicons from CGMMV-infected cucumber and watermelon plants and melon seeds were 99% identical to the CGMMV from the field melon. A cucumber plant infected with CGMMV but not SqMV was used for mechanical inoculation at the Contained Research Facility at University of California, Davis. Inoculated cucumber, melon, and watermelon plants showed green mottle and mosaic similar to that observed in the field. CGMMV is a highly contagious virus and damage by this virus on cucurbit crops has been reported in regions where CGMMV is present (2). CGMMV was detected on cucumber grown in greenhouses in Canada with 10 to 15% yield losses reported due to this virus (1). The three cucurbit crops in Yolo County were planted in an isolated area with no other cucurbits nearby. Measures, including destroying all the cucurbit plant material, have been taken to eradicate the virus. Use of CGMMV free cucurbit seed is necessary for prevention of this disease. References: (1) K.-S. Ling et al. Plant Dis. 98:701, 2014. (2) J. Y. Yoon et al. J. Phytopathol. 156:408, 2008.

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First Report of Tomato apical stunt viroid in Tomato in Tunisia

Plant Disease ◽

10.1094/pd-90-0528a ◽

2006 ◽

Vol 90 (4) ◽

pp. 528-528 ◽

Cited By ~ 9

Author(s):

J. Th. J. Verhoeven ◽

C. C. C. Jansen ◽

J. W. Roenhorst

Keyword(s):

Crop Production ◽

Plant Protection ◽

Chenopodium Quinoa ◽

Test Plant ◽

Production Facility ◽

Rt Pcr ◽

First Report ◽

Stunt Viroid ◽

Sequence Identity ◽

Two Samples

In May 2005, the Plant Protection Service in the Netherlands received two tomato (Lycopersicon esculentum) plant specimens for diagnosis from a protected crop production facility of 2.5 ha near Kebili in Tunisia. Growth of the plants was reduced and leaves were chlorotic and brittle. Ripening of the fruits was delayed and their storage life was reduced from 3 weeks to 1 week. The grower reported that initially only 5% of plants showed symptoms; however, the number of symptomatic plants increased quickly to 100% as a result of increasing temperatures in the production facility. Test plant species Chenopodium quinoa, Datura stramonium, Nicotiana glutinosa, N. hesperis-67A, N. occidentalis-P1, and L. esculentum ‘Money-maker’ were mechanically inoculated with sap from the affected plants. Symptoms including chlorosis and stunting were observed only on L. esculentum. Reverse transcriptase-polymerase chain reaction (RT-PCR) with universal pospiviroid primers Pospi1-RE/FW (2) yielded amplicons of the expected size (196 bp) for each of the two samples. One of these amplicons was sequenced and showed the highest identity to the four isolates of Tomato apical stunt viroid (TASVd) in the NCBI Gen-Bank. Subsequently, the complete sequence of the Tunisian isolate (Gen-Bank Accession No. DQ144506) was determined by sequencing the am-plicon obtained after RT-PCR using primers developed for the detection of Citrus exocortis viroid (CEVd) (1). The isolate consisted of 363 nucleotides and showed the highest sequence identity (96.7%) to tomato isolates of TASVd from Indonesia and Israel (GenBank Accession Nos. X06390 and AY062121, respectively), 92.6% to a tomato isolate from the Ivory Coast (GenBank Accession No. K00818), and 87.7% to an isolate from Solanum pseudocapsicum (GenBank Accession No. X95293). The next highest sequence identity was 81.5% to an isolate of CEVd (GenBank Accession No. X53716). On the basis of these results, the viroid was identified as TASVd. To our knowledge, this is the first report of TASVd in Tunisia. Reference: (1) N. Önelge. Turkish J. Agric. For. 21:419, 1997. (2) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004.

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First Report of Citrus bark cracking viroid and Citrus viroid V Infecting Citrus in China

Plant Disease ◽

10.1094/pdis-94-7-0922c ◽

2010 ◽

Vol 94 (7) ◽

pp. 922-922 ◽

Cited By ~ 6

Author(s):

M. J. Cao ◽

Y. Q. Liu ◽

X. F. Wang ◽

F. Y. Yang ◽

C. Y. Zhou

Keyword(s):

Poncirus Trifoliata ◽

Trifoliate Orange ◽

Rt Pcr ◽

First Report ◽

Local Cultivar ◽

Chongqing Municipality ◽

Rough Lemon ◽

Hop Stunt Viroid ◽

Production And Distribution ◽

One Step

Citrus is the most cultivated and highest value crop in the 15 southern provinces and municipalities in South China. Trifoliate orange (Poncirus trifoliata) is the main rootstock for citrus cultivars and is known to be susceptible to citrus viroids. Surveys conducted from 1995 to 2007 revealed 42 symptomatic samples from 33 cultivars (21 from sweet oranges (Citrus sinensis), 6 from mandarins (C. reticulata), 2 from satauma mandarins (C. unshiu), 6 from lemons (C. jambhiri), and 7 from mandarin hybrids). Symptoms included stunting, bark scaling, and cracking on the Trifoliate orange rootstock collected from citrus orchards in the Chongqing municipality, Sichuan, Zhejiang, Jiangxi, Hunan, and Yunnan provinces. Of the 42 samples, 27 were cultivars imported from abroad and 15 were local cultivars. Budwood from infected trees were grafted onto Arizona 861-S1 ‘Etrog citron’ (C. medica) on rough lemon (C. jambhiri) rootstock. After more than 12 months, 39 of 42 samples revealed typical viroid symptoms of stunting, epinasty. and leaf rolling on the Etrog indicator plants. In September 2009, total RNA was extracted with TRIZOL Reagent and a one-step multiplex reverse transcription (RT)-PCR assay (3) was used to detect simultaneously Citrus exocortis viroid (CEVd), Citrus bent leaf viroid (CBLVd), Hop stunt viroid (HSVd), and Citrus dwarfing viroid (CDVd). Also, a one-step RT-PCR protocol using two primer pairs targeting the complete genome sequences was used to detect Citrus bark cracking viroid (CBCVd) (1) and Citrus viroid V (CVd-V) (2). Of the 42 samples, 37 and 35 were positive for HSVd and CDVd, respectively. CEVd and CBLVd were found, respectively, in 14 and 13 of 42 samples. CBCVd was detected in cv. Meishan No. 9 (C. sinensis) from Sichuan Province and cvs. Akemi (C. reticulata) and Nishirokaori (C. reticulata) from Zhejiang Province. CVd-V was detected in cvs. Nishirokaori, Haruka (C. tamuranua), and Kiyomi (C. unshiu × C. sinensis) from Zhejiang, Hunan, and Chongqing Province, respectively. Only Meishan No. 9 is a local cultivar, whereas Akemi, Nishirokaori, Haruka, and Kiyomi are cultivars imported from Japan. Of 42 samples, 3 without typical symptoms on Etrog citrons were infected with HSVd only. Of 42 infected citrus plants, 36 harbored more than one viroid species. RT-PCR products of CBCVd and CVd-V were cloned by standard methods. Eight clones for CBCVd (one from Meishan No. 9 [Accession No. HM042742, 284 bp], three from Akemi [Accession Nos. HM042743-HM042745, 283 to 284 bp], and four from Nishirokaori [Accession Nos. HM042746-HM042749, 286 bp]) and six clones for CVd-V (four from Nishirokaori [Accession Nos. HM042750-HM042753, 294 bp], one from Kiyomi [Accession No. HM042754, 294 bp], and one from Haruka [Accession No. HM042755, 294 bp]) were sequenced and deposited in GenBank. BLAST analysis of the CBCVd (Accession No. HM042742) and CVd-V (Accession No. HM042751) sequences revealed highest nucleotide sequence identity (100 and 96%) to a CBCVd isolate from Cuba (Accession No. AJ630360) and a CVd-V isolate from Spain (Accession No. EF617306), respectively. To our knowledge, this is the first report of CBCVd and CVd-V in China. Our finding emphasizes the need for CBCVd and CVd-V indexing in production and distribution of pathogen-free citrus plants in China. References: (1) L. Bernard and N. Duran-Vila. Mol. Cell. Probes 20:105, 2006. (2) P. Serra et al. Phytopathology 98:1199, 2008. (3) X. F. Wang et al. Eur. J. Plant. Pathol. 124:175, 2009.

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First Report of Potato virus M and Chrysanthemum stunt viroid in Solanum jasminoides

Plant Disease ◽

10.1094/pd-90-1359a ◽

2006 ◽

Vol 90 (10) ◽

pp. 1359-1359 ◽

Cited By ~ 16

Author(s):

J. Th. J. Verhoeven ◽

C. C. C. Jansen ◽

J. W. Roenhorst

Keyword(s):

Plant Species ◽

Potato Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Test Plant ◽

Rt Pcr ◽

First Report ◽

Chrysanthemum Stunt Viroid ◽

Stunt Viroid ◽

Local Lesions ◽

Potato Virus M

In 2005, a plant of the ornamental crop Solanum jasminoides from the Netherlands was submitted for testing on viruses and viroids because of its intended use for propagation. Sap from this plant was mechanically inoculated to the test plant species Chenopodium quinoa, Nicotiana benthamiana, N. hesperis-67A, and N. occidentalis-P1 (3). N. hesperis-67A showed chlorotic local lesions and rugosity followed by vein necrosis, N. occidentalis-P1 showed necrotic local lesions and systemic leaf distortion, and the two other test plant species remained symptomless. Potato virus M (PVM) was identified by double antibody sandwich enzyme-linked immunosorbent assay using leaves from S. jasminoides and N. hesperis-67A. The plant of S. jasminoides was also tested for the presence of viroids by reverse transcriptase-polymerase chain reaction (RT-PCR) with universal pospiviroid primers Pospi1-RE/FW (2). This reaction yielded an amplicon of the expected size of 198 bp. The sequence showed 100% identity to an isolate of Chrysanthemum stunt viroid (CSVd; NCBI GenBank Accession No. AF394453). Subsequently, the complete sequence of our viroid isolate (GenBank Accession No. DQ406591) was determined from the amplicon obtained after RT-PCR using specific primers for the detection of CSVd (1). The viroid isolate from S. jasminoides consisted of 354 nucleotides and showed the highest identity (98.6%) to a chrysanthemum isolate of CSVd (GenBank Accession No. AB055974). Therefore, the viroid was identified as CSVd. To our knowledge, this is the first report of PVM and CSVd in S. jasminoides. Reference: (1) R. Hooftman et al. Acta Hortic. 432:120, 1996. (2) J. Th. J. Verhoeven et al. Eur. J. Plant Pathol. 110:823, 2004. (3) J. Th. J. Verhoeven and J. W. Roenhorst, EPPO Bull. 33:305, 2003.

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First Report of Tobacco rattle virus in Sedum in Minnesota

Plant Disease ◽

10.1094/pdis-94-3-0374a ◽

2010 ◽

Vol 94 (3) ◽

pp. 374-374 ◽

Cited By ~ 2

Author(s):

B. E. Lockhart ◽

S. L. Mason

Keyword(s):

Nucleotide Sequence ◽

Tobacco Rattle Virus ◽

Leaf Tissue ◽

Garden Soil ◽

Rt Pcr ◽

Electron Microscopic ◽

Arabis Mosaic Virus ◽

First Report ◽

Virus Like Particles ◽

Rigid Rod

Sedums (Sedum spp.; Crassulaceae) are perennial landscape plants that are grown widely because they are drought tolerant and winter hardy. Plants of Sedum ‘Matrona’ showing faint foliar ringspot symptoms were collected at a nursery retail outlet in St. Paul, MN in July 2008 and tested for possible viral infection by transmission electron microscopic (TEM) examination of negatively stained, partially purified leaf tissue extracts (1). The only virus-like particles observed were rigid, rod-shaped particles similar to those of Tobacco rattle virus (TRV) and other tobraviruses. A random sample of 100 measurements showed particles 20 nm in diameter with two modal lengths of 115 nm and 175 nm. These virus-like particles were confirmed to be those of TRV by immunosorbent electron microscopy (1) using antiserum to TRV (ATCC PVAS 75) and by reverse transcription (RT)-PCR using total RNA extracted with the RNeasy Kit (Qiagen, Valencia, CA) and primers that yield a 462-bp amplicon from TRV RNA 1 (4). An amplicon of the expected size was obtained by RT-PCR and its nucleotide sequence (GenBank Accession No. GQ268817) had 95 to 99% identity to published TRV sequences (AAW13192 and AAB48382). Two additional amplicons generated by RT-PCR from separate plants were identical in size and nucleotide sequence to the first. On the basis of virion morphology, serological relatedness, and sequence identity, the virus associated with mild ringspot symptoms in sedum was identified as an isolate of TRV. To our knowledge, this represents the first report of TRV incidence in sedum. Although Arabis mosaic virus is the only other virus reported to occur in sedum (2), we have observed numerous, flexuous filamentous 750 to 800 nm virus-like particles in partially purified extracts of a range of sedums showing mild mosaic and/or vein-clearing symptoms in Minnesota. Similar virus-like particles were not observed by TEM in partially purified extracts from TRV-infected ‘Matrona’ plants, suggesting that they did not contribute to the symptoms observed. We have reported previously (3) the occurrence of TRV in a variety of widely grown perennial ornamentals that provide potential sources of inoculum for spread of this virus by nematode vectors (Trichodorus and Paratrichodorus spp.) that occur commonly in garden soil, and Sedum is now added to the list of potential TRV reservoir plants. References: (1) Y. S. Ahlawat et al. Plant Dis. 80:590, 1996. (2) A. Gera et al. Acta Hortic. 722:175, 2006. (3) B. E. Lockhart et al. Plant Dis. 79:1249, 1995. (4) D. J. Robinson. J. Virol. Methods 40:57, 1992.

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Confirmation of First Report of Orchid fleck virus in Phalaenopsis Hybrid Orchids in the USA

Plant Health Progress ◽

10.1094/php-br-15-0018 ◽

2015 ◽

Vol 16 (4) ◽

pp. 146-148 ◽

Cited By ~ 3

Author(s):

S. A. Bratsch ◽

B. E. Lockhart ◽

C. Ishimaru

Keyword(s):

United States ◽

The United States ◽

Detection Methods ◽

Rt Pcr ◽

False Negatives ◽

Orchid Fleck Virus ◽

First Report ◽

The Usa ◽

One Step

The results also suggest one-step RT-PCR from Phalaenopsis plants and Orchid fleck virus (OFV) isolates can give false negatives for asymptomatic plants. Further work on detection methods for OFV is under way. This is the first confirmation of OFV infecting orchids in the United States. Accepted for publication 24 September 2015. Published 30 September 2015.

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First report of watermelon crinkle leaf-associated virus 1 (WCLaV-1) and WCLaV-2 in watermelon (Citrullus lanatus L.) plants co-infected with Cucurbit chlorotic yellows virus in Florida

Plant Disease ◽

10.1094/pdis-06-21-1141-pdn ◽

2021 ◽

Author(s):

Katherine Hendricks ◽

Regina Nicole Hernandez ◽

Pamela Roberts ◽

Thomas Isakeit ◽

Olufemi Joseph Alabi

Keyword(s):

Citrullus Lanatus ◽

South Florida ◽

Rt Pcr ◽

First Report ◽

Three Samples ◽

One Step ◽

Complete Coat Protein ◽

Species Specific ◽

Cucurbit Leaf Crumple Virus ◽

Cucurbit Chlorotic Yellows Virus

Watermelon (Citrullus lanatus L.) and other cucurbits are major crops in Florida. During the 2020 and 2021 seasons, watermelon plants with foliar virus-like symptoms of yellow mottling and chlorosis, mild leaf wrinkling and thickened leaves were observed in commercial fields (40 to 150 ha) in five counties (Desoto, Glades, Osceola, Seminole, and Charlotte) at >50% field incidence. Initial screening of 13 field-collected samples (2 to 4/County) for potyviruses with the Agdia POTY Immunostrip (Agdia, Inc. Elkhart, IN) were negative. Total nucleic acid extracts from each sample (RNeasy Plant Mini Kit, Qiagen, Germantown, MD) were used in one-step RT-PCR (Qiagen OneStep RT-PCR kit) with species-specific primer targeting squash vein yellowing virus (SqVYV), papaya ringspot virus-W (PRSV-W) (Adkins et al., 2008), cucurbit yellow stunting disorder virus (CYSDV) (Polston et al. 2008), cucurbit chlorotic yellows virus (CCYV) (Hernandez et al., 2021a), watermelon crinkle leaf-associated virus 1 (WCLaV-1), and WCLaV-2 (Hernandez et al., 2021b). The samples were also tested for cucurbit leaf crumple virus (CuLCrV) as per Hagen et al. (2008). All 13 samples were negative for SqVYV, PRSV-W, CuLCrV, and CYSDV, but 7 samples (53.8%) from 4 counties tested positive for CCYV, 12 (92.3%) from 5 counties were positive for WCLaV-1, and WCLaV-2 was detected in 8 samples (61.5%) from 5 counties. Three samples were singly infected with WCLaV-1 while the remaining 10 were mixed infected with different combination of 2 or 3 viruses. Notably, symptoms on all 13 plants were visually indistinguishable. To verify the results, two randomly chosen gene-specific fragments per virus, obtained with primers CCYV-v1330/c2369, CCYV-v4881/c5736, WCLaV-1vRP/1cRP, WCLaV-1vMP/1cMP, WCLaV-2vRP/2cRP, and WLaV 2vMP/2cMP, were excised from the gel, cloned, and Sanger-sequenced as described (Hernandez et al., 2021a, 2021b). In pairwise comparisons, the ~1 kb partial ORF1a (GenBank accession nos. MZ325846 to MZ325847) and 753 bp complete coat protein cistron (MZ325848 to MZ325849) of CCYV from Florida shared 98.9-99.5%/98.4-99.6% nucleotide (nt)/amino acid (aa) and 99.4-99.8%/99.6-100% nt/aa identities, respectively with the corresponding sequences of global CCYV isolates. The partial RNA1 (MZ325850 to MZ325851) and RNA2 (MZ325852 to MZ325853) sequences of WCLaV-1 from Florida shared 99.2-99.8%/100% nt/aa and 98.9-100%/99.3-100% nt/aa identities, respectively with the corresponding global sequences of WCLaV-1 isolates. Lastly, the partial RNA1 (MZ325854 to MZ325857) and RNA2 (MZ325858 to MZ325861) sequences of WCLaV-2 from Florida shared 96.4-99.8%/97-100% nt/aa and 96.5-100%/95.9-100% nt/aa identities, respectively with the corresponding global sequences of WCLaV-2 isolates. This is the first report of WCLaV-1 and WCLaV-2 from Florida and the first documentation of the occurrence of CCYV in South Florida. CCYV has been reported previously from California (Wintermantel et al. 2019), Georgia (Kavalappara et al. 2021), and recently from North Florida (M. Paret, pers. comm) but WCLaV-1 and WCLaV-2 have only been reported from Texas (Hernandez et al., 2021b), after their discovery in China (Xin et al. 2017). The results indicate further expansion of the geographical range of these cucurbit-infecting viruses, although their longer but undetected presence in Florida is plausible due to the resemblance of their associated symptoms with those attributed to known viruses. References Adkins, et al., 2008. Plant Dis. 92:1119-1123. https://apsjournals.apsnet.org/doi/10.1094/PDIS-92-7-1119. Hagen, et al. 2008. Plant Dis. 92:781-793. https://apsjournals.apsnet.org/doi/pdfplus/10.1094/PDIS-92-5-0781. Hernandez, et al. 2021a. Plant Dis. https://doi.org/10.1094/PDIS-02-21-0378-PDN. Hernandez, et al., 2021b. Plant Dis. https://doi.org/10.1094/PDIS-02-21-0249-PDN Kavalappara, et al., 2021. Plant Dis. https://doi.org/10.1094/PDIS-11-20-2429-PDN. Polston, et al. 2008. Plant Dis. 92(8):1251. https://apsjournals.apsnet.org/doi/10.1094/PDIS-92-8-1251B. Wintermantel, et al., 2019. Plant Dis. 103(4):778. https://doi.org/10.1094/PDIS-08-18-1390-PDN. Xin, et al., 2017. Front. Microbiol. 8:1514, doi: 10.3389/fmicb.2017.01514.

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