scholarly journals First Report of Potato mop-top virus on Potato from the United States

Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 872-872 ◽  
Author(s):  
D. H. Lambert ◽  
L. Levy ◽  
V. A. Mavrodieva ◽  
S. B. Johnson ◽  
M. J. Babcock ◽  
...  
Keyword(s):  
Triple Gene Block ◽  
The United States ◽  
Russet Burbank ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coding Region ◽  
Rt Pcr ◽  
Protein Coding ◽  
Gene Block ◽  
Elisa Testing ◽  
Potato Mop Top Virus

Potato mop-top virus (PMTV) is a tripartite pomovirus vectored by the powdery scab plasmodiophoromycete Spongospora subterranea pv. subterranea (1). PMTV occurs on potato (Solanum tuberosum) in Europe, the Andes, Asia, and Canada. Internal necrotic arc and fleck tuber symptoms (“spraing”) may reduce commercial acceptance of some cultivars (3). PMTV symptoms were discovered in ‘Shepody’ tubers at the Aroostook Research Farm, Presque Isle, ME in May 2002 and subsequently in ‘Russet Burbank’ tubers in commercial storage from the 2001 Maine crop. Symptomatic tubers exhibited single or multiple concentric necrotic arcs that were partial or complete, but exhibited no distinct external symptoms. The presence of PMTV in eight ‘Shepody’ tubers was indicated by positive enzyme-linked immunosorbent assay (ELISA; Adgen, Ltd., Auchincruive, Ayr, Scotland) and confirmed by reverse transcription polymerase chain reaction (RT-PCR). ‘Russet Burbank’ potatoes were visually diagnosed, and the corresponding halves of 128 symptomatic tubers were forwarded to the University of Maine and APHIS (Beltsville, MD). Of these, ELISA readings in Maine were strongly positive (>3 × background) for 88, ambiguous (1.5-3 × background) for 13, and negative for 27. Subsamples from these three categories were positive by PCR in 17 of 17, 9 of 9, and 12 of 14 cases, respectively. A similar rating, positive or ambiguous, in ELISA testing was identical for all but one case at Beltsville. Confirmation of PMTV required PCR testing, resulting in a characteristic PCR product of 401 bp that was generated from the coat protein coding region on RNA 2 (2) using the primer pair PMTV 1 5′-GCAGCCGTCGAGAATAGATA-3′ (RNA nucleotides 316–335) and PMTV 4 5′-GCGAGTTGATGTGCC ACATT-3′ (complementary to RNA 2 nucleotides 716–697). An immunocapture RT-PCR using this primer set and the coating antibody from the Adgen ELISA kit was also successful in detecting PMTV. In separate reactions, a second product of 646 bp was generated from the triple gene block on RNA 3 (4) using the primer pair PMTV 5 5′-GGTGAACACGAGGACAAGGT-3′ (RNA 3 nucleotides 1417–1436) and PMTV 7 5′-AACAGTCCGGTCTTGTGAAC-3′ (complementary to RNA 3 nucleotides 2063–2044). The sequence of these products was 98 to 100% identical to PMTV published sequences. The discovery of this virus will result in adjustments to U.S. and Canadian seed potato certification standards and symptom characterization for common North American cultivars. References: (1) R. A. C. Jones and B. D. Harrison. Ann. Appl. Biol 63:1, 1969. (2) S. Kashiwazak et al. Virology 206:701, 1995. (3) M. Sandgren et al. Am. J. Potato Res. 79:205, 2002. (4) K. P. Scott et al. J. Gen. Virol.75:3561, 1994.

scholarly journals Detection and Confirmation of Potato mop-top virus in Potatoes Produced in the United States and Canada

Plant Disease ◽  
2004 ◽  
Vol 88 (4) ◽  
pp. 363-367 ◽  
Author(s):  
H. Xu ◽  
T.-L. DeHaan ◽  
S. H. De Boer
Keyword(s):  
United States ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
The North ◽  
Potato Mop Top Virus ◽  
Pcr Targeting

Potato mop-top virus (PMTV) was detected in potatoes grown in the United States and Canada during surveillance testing by a reverse transcription-polymerase chain reaction (RT-PCR) targeting the coat protein gene in RNA3. Out of 3,221 lots of seed and ware potatoes that were tested, 4.3% were positive for PMTV. The reliability of the survey results was confirmed by reextraction of selected samples and additional RT-PCR tests using two primer sets targeting gene segments in RNA2 and RNA3. Amplicons generated from RNA2 and RNA3 were identified by analysis of fragment length polymorphisms after digestion with BamHI and HindIII, respectively. PMTV was further identified by enzyme-linked immunosorbent assay, bioassay on Nicotiana debneyi, and transmission electron microscopy. Sequencing of a portion of the coat protein gene revealed near 100% identity among isolates from the United States and Canada and >97% homology of the North American isolates with European isolates.

scholarly journals First Report of Grapevine leafroll-associated virus 2 in a Hybrid Grape ‘Vidal Blanc’ in Missouri

Plant Disease ◽  
2012 ◽  
Vol 96 (3) ◽  
pp. 462-462 ◽  
Author(s):  
S. Lunden ◽  
W. Qiu
Keyword(s):  
New York ◽  
Leaf Roll ◽  
Late Summer ◽  
The United States ◽  
Coding Region ◽  
Rt Pcr ◽  
Protein Coding ◽  
First Report ◽  
Stem Lesions ◽  
Grape Cultivars

Grapevine leaf roll disease (GLRD) is one of the most prevalent viral diseases in vineyards worldwide. At least 10 Grapevine leafroll-associated viruses (GLRaV-1 to -7, and -9, -10, and -11) are associated with GLRD. GLRaV-2 has a number of distinct isolates that are associated not only with GLRD, but also with graft incompatibility, young vine decline, and rootstock stem lesions. In the United States, GLRaV-2 isolates have been reported in the states of California (4), New York (1), Oregon, and Washington (3). A survey of grapevine viruses was conducted on seven grape cultivars, Norton, Chambourcin, Chardonel, Vignoles, Vidal Blanc, Traminette, and Cayuga White, in a vineyard located at Mountain Grove, MO during the falls of 2009 and 2010 and late summer of 2011. Vines from each cultivar were assigned into three blocks. From each block, six individual vines were randomly selected. Phloem scrapings were collected for detecting viruses by ELISA and reverse-transcription (RT)-PCR. To test for GLRaV-2, a set of primers, GLRaV-2-Forward (5′-GGTGATAACCGACGCCTCTA, nt 6745 to 6764) and GLRaV-2-Reverse (5′-CCTAGCTGACGCAGATTGCT, nt 7268 to 7287), were designed from the coat protein coding region as reported previously (2). GRLaV-2 was detected only in the samples of ‘Vidal Blanc’. The detection of GLRaV-2 was further verified by ELISA using GRLaV-2-specific antisera (Bioreba AG, Reinach, Switzerland). The GLRaV-2-infected ‘Vidal Blanc’ vines did not exhibit visible symptoms. The RT-PCR amplified 543-bp cDNA fragments were isolated from agarose gel and cloned into the pCR2.1 vector (Invitrogen, Carlsbad, CA). Plasmid DNA was purified from three individual clones and sequenced from both directions. Comparison of the 543-bp sequences showed that the sequences shared 99% nt identity with the corresponding regions of 21 GLRaV-2 isolates, including PMC-083 isolate from Croatia (GenBank Accession No. HM185277), LN isolate from China (GenBank No. FJ786017), SE isolate from Brazil (GenBank No. EU204909), and four isolates from Washington (GenBank Nos. EU760836, 760838, 760843, and 760848). ‘Vidal Blanc’ is a white interspecific hybrid grape. Absence of visible symptoms on this cultivar emphasizes the importance of planting certified grapevines that have been indexed for viruses. Discovery of GLRaV-2 in asymptomatic white grape cultivars will warn growers on the potential damages and epidemics of GLRD in Midwest vineyards and promote planting clean grapevines. To our knowledge, this is the first report of GLRaV-2 in grapevines in Missouri. References: (1) M. Fuchs et al. Plant Dis. 93:395, 2009. (2) G. Gambino and I. Gribaudo. Phytopathology 96:1223, 2006. (3) R. R. Martin et al. Plant Dis. 89:763, 2005. (4) A. Rowhani et al. Phytopathology (Abstr.) 92:(suppl.):S71, 2002.

scholarly journals Detection of Potato mop top virus (Furovirus) on potato in southeast Idaho

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 149-149 ◽  
Author(s):  
J. L. Whitworth ◽  
J. M. Crosslin
Keyword(s):  
North Dakota ◽  
Tobacco Rattle Virus ◽  
The United States ◽  
Positive Control ◽  
Russet Burbank ◽  
Rt Pcr ◽  
Water Control ◽  
Total Rna ◽  
Potato Mop Top Virus ◽  
Multiple Samples

In February 2012, commercially produced potato (Solanum tuberosum) tubers, cv. Modoc, grown in southeast Idaho, were observed with internal necrotic arcs and lines. Samples were obtained from potatoes that had been washed and packaged. No external symptoms were evident. Multiple samples were collected from the packing line and cut to check for internal defects as part of the normal grading procedure. The incidence of symptomatic tubers from these samples was determined by personnel at the packaging facility to be approximately 3%. Initially, one symptomatic and one asymptomatic tuber were tested. Total RNA extracted from these tubers were tested by RT-PCR at Aberdeen, Idaho, with primers specific for Potato mop top virus (PMTV) (1) and Tobacco rattle virus (TRV) (4). RT-PCR results showed that the symptomatic tuber produced a band at 416 bp with the PMTV primers, which was also present in the PMTV-positive control. No amplification was observed with the TRV primers. The asymptomatic tuber was negative for both PMTV and TRV. Subsequently, total RNA from four additional symptomatic tubers from the same lot were tested at USDA-ARS in Prosser, WA, by RT-PCR for TRV (4) and with a different set primers for PMTV (2). The tests included two PMTV-positive controls from cv. Alturas tubers (1), a healthy cv. Russet Burbank control, and a water control. Results showed that amplified products of 460 bp were obtained with the PMTV primers for the four symptomatic tubers and the same tubers were negative for TRV. In addition, symptomatic tissue from the four tubers tested positive for PMTV by ELISA using a commercially available kit (Adgen, Ayr, Scotland). Symptomless Russet Burbank tubers and water controls were negative in RT-PCR and ELISA tests. The 460 bp PMTV amplicon from two symptomatic Modoc tubers were cloned and sequenced. The sequences were identical and the sequence (GenBank Accession No. JX239990) was 100% identical to the corresponding sequences of PMTV isolates from North Dakota (HM776172) and Finland (AM503632). There was one nucleotide difference from the corresponding sequence of a PMTV isolate from Washington (JN132117). To our knowledge, this is the first published report of PMTV in Idaho and confirms that PMTV exists in southeast Idaho. A previous report made by Canada in 2004 (Plant Dis. 88:363) indicates that PMTV was found in multiple states and provinces, but no specific locations were identified. This report follows reports of PMTV in commercial potatoes in Washington (1), North Dakota (2), and Maine (3). In 2011, 129,000 hectares of potatoes were grown in Idaho, representing 29% of the fall grown potatoes in the United States. PMTV can cause quality problems and as evidenced by these samples with no external symptoms, problems may be compounded because of internal symptoms that may go undetected. The confirmation of PMTV alerts growers and processors to the presence of this virus in this important potato-producing state. References: (1) J. M. Crosslin. Plant Dis. 95:1483, 2011. (2) N. David et al.Plant Dis. 94:1506, 2010. (3) D. H. Lambert et al.Plant Dis. 87:872, 2003. (4) D. J. Robinson. J. Virol. Methods 40:57, 1992.

scholarly journals Epidemiological Survey and Screening Strategy for Dengue Virus in Blood Donors from Yunnan Province

2020 ◽  
Author(s):  
LING LI ◽  
YING LI ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Prevalence Rate ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Igm Antibodies ◽  
Blood Center ◽  
Nonstructural Protein 1 ◽  
Elisa Testing

Abstract BACKGROUND Dengue virus (DENV) can be transmitted through blood transfusion. DENV was not screened regularly in Xishuangbanna Blood Center. This study was conducted in Xishuangbanna Blood Center with an attempt to develop DENV screening strategies in one of China’s high-incidence areas.METHODS Blood samples were collected randomly between June 2019 and August 2019. These samples were first screened for dengue IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). All reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real time polymerase-chain-reaction (RT-PCR) assay. After RT-PCR assay, these samples were further tested for soluble nonstructural protein 1 (NS1) using colloidal gold method. The demographic data of DENV positive donors were collected.RESULTS A total of 2,254 donor samples were collected and tested for dengue IgG and IgM antibodies by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-IgG and/or anti-IgM reactive, with a serological prevalence rate of 26.53%. Among all the donor samples, 26 were RT-PCR positive and/or NS1 positive. Moreover, there were significant differences in the prevalence rate of DENV in terms of occupation (P=0.001), education(P<0.001) and ethnicity (P=0.026). CONCLUSION The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides the first-hand data about the prevalence of DENV and allows development of a screening strategy for clinical use.

scholarly journals First Report of Lolium latent virus in Ryegrass in the United States

Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 528-528 ◽  
Author(s):  
C. J. Maroon-Lango ◽  
J. Hammond ◽  
S. Warnke ◽  
R. Li ◽  
R. Mock
Keyword(s):  
United States ◽  
Lolium Perenne ◽  
The United States ◽  
Latent Virus ◽  
Hybrid Population ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
High Sequence Identity ◽  
Lolium Perenne L ◽  
Pcr Test

Initial reports of the presence of Lolium latent virus (LLV) in Lolium perenne L. and L. multiflorum Lam. breeding clones in Germany, the Netherlands, France (2), and recently the United Kingdom (3,4; described as Ryegrass latent virus prior to identification as LLV) prompted us to evaluate clonally propagated Lolium plants from the United States. Four genetically distinct plants (viz., MF22, MF48, MF125, and MF132) that have been maintained clonally for 5 years from a Lolium perenne × L. multiflorum hybrid population established in the United States exhibited either no symptoms or mild chlorotic flecking that coalesced to form chlorotic to necrotic streaking on the leaves. All four clonal plants tested positive using reverse transcription-polymerase chain reaction (RT-PCR) with the Potexvirus group PCR test (Agdia, Inc., Elkhart, IN), whereas all clones but MF48 tested positive using the Potyvirus group PCR test (Agdia, Inc.). No amplicons were obtained when the same plants were tested for tobamovirus, carlavirus, and closterovirus using appropriate virus group-specific primers. Cloning and sequencing of the potexviral amplicons revealed very high sequence identity with the comparable region of LLV-UK (GenBank Accession No. DQ333886), whereas those of the potyviral amplicons (GenBank Accession Nos. DQ355837 and DQ355838) were nearly identical with the comparable region of Ryegrass mosaic virus (RGMV), a rymovirus first reported from the United States in 1957 (1). Using indirect enzyme-linked immunosorbent assay (ELISA), extracts from all four Lolium clonal propagations tested positive for LLV using the antiserum raised to LLV-Germany (courtesy of Dr. Huth), whereas the potyvirus-positive results from RT-PCR of the three clones were confirmed using indirect ELISA with the broad spectrum potyvirus monoclonal antibody, PTY-1. LLV from singly or dually infected Lolium clones was transmitted to Nicotiana benthamiana Domin. but not to N. tabacum L. by mechanical inoculation. LLV was purified from infected N. benthamiana. Similar sized flexuous rods were observed using electron microscopy in leaf dip samples from Lolium clones and aliquots of the virions purified from N. benthamiana. References: (1) G. W. Bruehl et al. Phytopathology 47:517, 1957. (2) W. Huth et al. Agronomie 15:508, 1995. (3) R. Li et al. Asian Conf. Plant Pathol. 2:89, 2005. (4) C. Maroon-Lango et al. Int. Congr. Virol. 13:63, 2005.

scholarly journals First Report of Grapevine leafroll associated virus-3 in American Vitis spp. Grapevines in Washington State

Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1461-1461 ◽  
Author(s):  
M. J. Soule ◽  
K. C. Eastwell ◽  
R. A. Naidu
Keyword(s):  
New York ◽  
Economic Impact ◽  
Virus Disease ◽  
The United States ◽  
Washington State ◽  
The State ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Table Grapes

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

scholarly journals First Report of Plum pox virus (Sharka Disease) in Prunus persica in the United States

Plant Disease ◽  
2000 ◽  
Vol 84 (2) ◽  
pp. 202-202 ◽  
Author(s):  
L. Levy ◽  
V. Damsteegt ◽  
R. Welliver
Keyword(s):  
Prunus Persica ◽  
Virus Disease ◽  
Sandwich Elisa ◽  
Pcr Amplification ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Peach Fruit ◽  
First Report

Plum pox (Sharka) is the most important virus disease of Prunus in Europe and the Mediterranean region and is caused by Plum pox potyvirus (PPV). In September 1999, PPV-like symptoms were observed in peach fruit culls in a packinghouse in Pennsylvania. All symptomatic fruit originated from a single block of peach (P. persica cv. Encore) in Adams County. Trees in the block exhibited ring pattern symptoms on their leaves. A potyvirus was detected in symptomatic fruit using the Poty-Group enzyme-linked immunosorbent assay (ELISA) test from Agdia (Elkhart, IN). Reactions for symptomatic peach fruit and leaves also were positive using triple-antibody sandwich ELISA with the PPV polyclonal antibody from Bioreba (Carrboro, NC) for coating, the Poty-Group monoclonal antibody (MAb; Agdia) as the intermediate antibody, and double-antibody sandwich ELISA with PPV detection kits from Sanofi (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) and Agdia and the REAL PPV kit (Durviz, Valencia, Spain) containing universal (5B) and strain typing (4DG5 and AL) PPV MAbs (1). PPV also was identified by immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) amplification and subsequent sequencing of the 220-bp 3′ noncoding region (2) (>99% sequence homology to PPV) and by IC-RT-PCR amplification of a 243-bp product in the coat protein (CP) gene (1). The virus was identified as PPV strain D based on serological typing with strainspecific MAbs and on PCR-restriction fragment length polymorphism of the CP IC-RT-PCR product with Rsa1 and Alu1 (1). This is the first report of PPV in North America. References: (1) T. Candresse et al. Phytopathology 88:198, 1998. (2) L. Levy and A. Hadidi. EPPO Bull. 24:595, 1994.

scholarly journals Alternanthera mosaic virus Found in Scutellaria, Crossandra, and Portulaca spp. in Florida

Plant Disease ◽  
2006 ◽  
Vol 90 (6) ◽  
pp. 833-833 ◽  
Author(s):  
C. A. Baker ◽  
L. Breman ◽  
L. Jones
Keyword(s):  
Electron Microscopy ◽  
United States ◽  
Mosaic Virus ◽  
Plant Species ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Indicator Plants ◽  
Partial Identity ◽  
Pcr Products

In the fall of 1998, the Division of Plant Industry (DPI) received vegetative propagations of Scutellaria longifolia (skullcap) with symptoms of foliar mosaic, chlorotic/necrotic ringspots, and wavy line patterns from a nursery in Manatee County. Flexuous particles approximately 500 nm long were found with electron microscopy. The plants tested positive for Papaya mosaic virus (PaMV) in an enzyme-linked immunosorbent assay (ELISA) test with antiserum to PaMV (Agdia, Elkhart, IN). However, in immunodiffusion tests (antiserum from D. Purcifull, University of Florida), this virus gave a reaction of partial identity indicating it was related but not identical to PaMV (1). The original infected plants were kept in a greenhouse. In January 2005, a specimen of Crossandra infundibuliformis (firecracker plant) with mosaic symptoms was submitted to the DPI from a nursery in Alachua County. Inclusions found with light microscopy and particles found with electron microscopy indicated that this plant was infected with a potexvirus. This was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) with primers designed to detect members of the virus family Potexviridae (3). These plants reacted positive to PaMV antiserum in ELISA and gave a reaction of partial identity to PaMV in immunodiffusion. A specimen of Portulaca grandiflora (moss rose) with distorted leaves found at a local retail store was also tested and gave the same results. Leaves from each of the three plant species were rubbed onto a set of indicator plants using Carborundum and potassium phosphate buffer. Total RNA was extracted from symptomatic indicator plants of Nicotiana benthamiana. RT-PCR (3) was performed, and PCR products were sequenced directly. Sequences of approximately 700 bp were obtained for all three plant species and showed 98% identity with each other. BLAST search results showed that these sequences were 93% identical to an Alternanthera mosaic virus (AltMV) sequence at the nucleotide level but only 76% identical to PaMV. The amino acid sequences were 98 and 82% identical to AltMV and PaMV, respectively. The PCR products of the virus from Scutellaria sp. were cloned, resequenced, and the sequence was entered into the GenBank (Accession No. DQ393785). The bioassay results matched those found for AltMV in Australia (2) and the northeastern United States (4), except that the Florida viruses infected Datura stramonium and Digitalis purpurea (foxglove). The virus associated with the symptoms of these three plants appears to be AltMV and not PaMV. AltMV has been found in ornamental plants in Australia, Italy, and the United States (Pennsylvania, Maryland, and now Florida). Since this virus is known to infect several plants asymptomatically and can be easily confused with PaMV serologically, it is likely that the distribution of this virus is much wider than is known at this time. References: (1) L. L. Breman. Plant Pathology Circular No. 396. Fla. Dept. Agric. Consum. Serv. DPI, 1999. (2) A. D. W. Geering and J. E. Thomas. Arch Virol 144:577, 1999. (3) A. Gibbs et al. J Virol Methods 74:67, 1998. (4) J. Hammond et al. Arch Virol. 151:477, 2006.

scholarly journals Transient Coexpression of Individual Genes Encoded by the Triple Gene Block of Potato mop-top virus Reveals Requirements for TGBp1 Trafficking

2004 ◽  
Vol 17 (8) ◽  
pp. 921-930 ◽  
Author(s):  
Andrey A. Zamyatnin ◽  
Andrey G. Solovyev ◽  
Eugene I. Savenkov ◽  
Anna Germundsson ◽  
Maria Sandgren ◽  
...  
Keyword(s):  
Cell Wall ◽  
Fluorescent Protein ◽  
Triple Gene Block ◽  
Terminal Region ◽  
Virus Movement ◽  
Cell Periphery ◽  
Movement Proteins ◽  
Gene Block ◽  
Potato Mop Top Virus ◽  
Green Fluorescent

TGBp1, TGBp2, and TGBp3, three plant virus movement proteins encoded by the “triple gene block” (TGB), may act in concert to facilitate cell-to-cell transport of viral RNA genomes. Transient expression of Potato mop-top virus (genus Pomovirus) movement proteins was used as a model to reconstruct interactions between TGB proteins. In bombarded epidermal cells of Nicotiana benthamiana, green fluorescent protein (GFP)-TGBp1 was distributed uniformly. However, in the presence of TGBp2 and TGBp3, GFP-TGBp1 was directed to intermediate bodies at the cell periphery, and to cell wall-embedded punctate bodies. Moreover, GFP-TGBp1 migrated into cells immediately adjacent to the bombarded cell. These data suggest that TGBp2 and TGBp3 mediate transport of GFP-TGBp1 to and through plasmodesmata. Mutagenesis of TGBp1 suggested that the NTPase and helicase activities of TGBp1 were not required for its transport to intermediate bodies directed by TGBp2 and TGBp3, but these activities were essential for the protein association with cell wall-embedded punctate bodies and translocation of TGBp1 to neighboring cells. The C-terminal region of TGBp1 was critical for trafficking mediated by TGBp2 and TGBp3. Mutation analysis also suggested an involvement of the TGBp2 C-terminal region in interactions with TGBp1.

scholarly journals Characterization of Tomato Necrotic Spot Virus, a Subgroup 1 Ilarvirus Causing Necrotic Foliar, Stem, and Fruit Symptoms in Tomatoes in the United States

Plant Disease ◽  
2019 ◽  
Vol 103 (6) ◽  
pp. 1391-1396 ◽  
Author(s):  
Sara A. Bratsch ◽  
Samuel Grinstead ◽  
Tom C. Creswell ◽  
Gail E. Ruhl ◽  
Dimitre Mollov
Keyword(s):  
Genomic Sequence ◽  
The United States ◽  
Amino Acid Identity ◽  
Necrotic Spot ◽  
Coding Region ◽  
Spot Virus ◽  
Protein Coding ◽  
Serological Characterization ◽  
Necrotic Spots

The genomic, biological, and serological characterization of tomato necrotic spot virus (ToNSV), a virus first described infecting tomato in California, was completed. The complete genomic sequence identified ToNSV as a new subgroup 1 ilarvirus distinct from the previously described tomato-infecting ilarviruses. We identified ToNSV in Indiana in 2017 and 2018 and in Ohio in 2018. The coat protein coding region of the isolates from California, Indiana, and Ohio have 94 to 98% identity, while the same isolates had 99% amino acid identity. ToNSV is serologically related to TSV, a subgroup 1 ilarvirus, and shows no serological relationship to ilarviruses in the other subgroups. In tomato, ToNSV caused symptoms of necrotic spots and flecks on leaves, necrotic streaking on stems, and necrotic spots and circular patterns on fruit resulting in a yield loss of 1 to 13%. These results indicate that ToNSV is a proposed new subgroup 1 ilarvirus causing a necrotic spotting disease of tomato observed in California, Indiana, and Ohio.

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