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 Three Previously Unrecorded Viral Diseases of Astilbe, Fuschia, and Thermopsis Species in Minnesota

Plant Disease ◽  
2005 ◽  
Vol 89 (7) ◽  
pp. 775-775 ◽  
Author(s):  
B. E. L. Lockhart
Keyword(s):  
Electron Microscopy ◽  
Mosaic Virus ◽  
The United States ◽  
Yellow Mosaic Virus ◽  
Viral Diseases ◽  
Enzyme Linked Immunosorbent Assay ◽  
Leaf Extracts ◽  
Indicator Plants ◽  
Virus Like Particles ◽  
Leaf Deformation

Interest in virus diseases of perennial ornamentals has been increasing because of their increasing monetary value, because wholesale producers perceive an advantage in marketing disease-free stock, and because widespread international movement of these plants carries the risk of introduction of exotic viruses. In an ongoing study to identify and document viral diseases of perennial ornamentals used in the United States commercial horticultural industry, three virus-like diseases of astilbe (Astilbe chinensis), fuschia (Fuschia cv. Gartenmeister) and false lupine (Thermopsis caroliniana) occurring in Minnesota were investigated. Symptomatic plants were selected from lots in commercial greenhouses and garden centers in several locations in Minnesota. Astilbe with systemic chlorosis was found to be infected with Tobacco ringspot virus (TRSV). Fuschia with leaf mottling and leaf deformation was found to be infected with Cucumber mosaic virus (CMV), and false lupine with mosaic and leaf deformation symptoms was found to be infected with Bean yellow mosaic virus (BYMV). Identification of the three viruses was based on: 1) virion presence and morphology in partially purified leaf extracts using electron microscopy (EM) and immunosorbent electron microscopy (1); 2) enzyme-linked immunosorbent assay using crude leaf extracts; and 3) biological properties, including symptoms produced in indicator plants. Antisera to BYMV (ATCC PVAS-368), CMV (ATCC PVAS-30), and TRSV (ATCC PVAS-157) were obtained from the American Type Culture Collection, Manassas, VA. No other virus-like particles were observed with EM in partially purified leaf extracts of the three plants, no virus-like particles were observed in similar preparations from asymptomatic plants, and indicator plant tests did not indicate the presence of any other mechanically transmissible viruses. The TRSV isolate from astilbe and the BYMV isolate from false lupine produced typical symptoms on indicator plants susceptible to known isolates of these two viruses. The CMV isolate from fuschia was similar to previously described isolates of CMV (2) in most respects and was readily transmitted in a nonpersistent manner by Myzus persicae, but was unusual in that it did not infect Nicotiania benthamiana, N. glutinosa, and tomato, which are normally highly susceptible to infection by CMV. The identity of the fuschia CMV isolate was further confirmed by reverse transcription-polymerase chain reaction (RT-PCR) amplification with CMV-specific oligonucleotide primers (3). The PCR product was of the predicted size (500 bp) and was cleaved by restriction digestion with EcoRI, suggesting that the fuschia virus is a Type II CMV isolate (3). To my knowledge, this is the first report of TRSV infection in astilbe, CMV infection in fuschia, and of a viral disease of false lupine. References: (1) Y. C. Ahlawat et al. Plant Dis. 80:590, 1996. (2) A. A. Brunt et al. Viruses of Plants. CAB International, Wallingford, UK, 1995. (3) S. Wylie et al. Aust. J. Agric. Res. 44:41, 1993.

scholarly journals First Report of Alfalfa mosaic virus Occurrence in Hydrangea in the United States

Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1258-1258 ◽  
Author(s):  
B. Lockhart ◽  
D. Mollov ◽  
M. Daughtrey
Keyword(s):  
Electron Microscopy ◽  
United States ◽  
Mosaic Virus ◽  
Alfalfa Mosaic Virus ◽  
Leaf Tissue ◽  
The United States ◽  
Spherical Particles ◽  
Genomic Sequences ◽  
Rt Pcr ◽  
First Report

In spring of 2012, a previously unrecorded virus-like disease characterized by conspicuous yellow leaf blotching (calico symptoms) was observed in plants of Hydrangea macrophylla in a single location in Southampton, NY. Bacilliform and spherical particles resembling those of Alfalfa mosaic virus (AMV) were observed by transmission electron microscopy (TEM) in partially purified extracts from symptomatic leaf tissue. The identity of the virus was confirmed by immunosorbent electron microscopy (ISEM) (4) using antiserum to AMV (ATCC PVAS 92) that both trapped and decorated the virions. Three primer pairs designed from available AMV RNA 1, RNA 2, and RNA 3 genomic sequences were used to generate amplicons from the hydrangea AMV isolate. Reverse-transcription (RT)-PCR was done using total RNA extracted from symptomatic hydrangea leaf tissue with a Qiagen RNeasy kit, and Ready-to-Go RT-PCR beads (GE Healthcare). Amplicons of 1,049, 1,013, and 658 bp were obtained using the primer pairs AMV1F (5′-ATCCACCGATGCCAGCCTTA)/AMV1R (5′-TTCCGCCTCACTGCTGTCTG), AMV2F (5′-GATCGCCGGAAGTGATCCAG)/AMV2R (5′-TCACCGGAAGCAACAACGAA), and AMV3F (5′-GCCGGTTCTCCAAAGGGTCT)/AMV3R (5′-CGCGTCGAAGTCCAGACAGA), respectively. The PCR products were cloned using a TOPO TA cloning kit (Invitrogen) and three clones of each were sequenced. The sequences obtained from the hydrangea AMV RNA 1 (JX154090), RNA 2 (JX154091), and RNA 3 (JX154092) had 95 to 98% nucleotide sequence identity to homologous genomic sequences of known AMV isolates. To our knowledge, this is the first report of AMV occurrence in H. macrophylla in the United States. This virus has been reported to occur in H. macrophylla in British Columbia (3), but in a previous survey its presence was not detected in hydrangeas in the United States (1). A report of possible AMV infection in H. macrophylla in Italy (2) was based solely on symptomatology and cross-protection tests and therefore cannot be verified. The AMV-infected hydrangea plants were found by ISEM to also contain low concentrations of Hydrangea ringspot virus (HRSV) and Hydrangea chlorotic mottle virus (HdCMV). However, based on previous evidence of single and mixed infections (3), it is unlikely that the calico symptoms observed were influenced by the presence of HRSV and HdCMV. This report is of interest both because AMV, unlike HRSV and HdCMV, causes foliar symptoms that would render hydrangea plant unmarketable, and because the disease can be spread by a number of common aphid species that transmit AMV. It will also serve to alert growers and diagnosticians to the potential threat posed by AMV infection. References: (1) T. C. Allen et al. Acta Hortic. 164:85, 1985. (2) G. Belli. Phytopathol. Mediterr. 7:70, 1968. (3) A. W. Chiko and S. E. Godkin. Plant Dis. 70:541, 1986. (4) B. E. L. Lockhart et al. Phytopathology 82:691, 1992.

scholarly journals First Report of Cucumber mosaic virus Infection in Pachysandra in the United States

Plant Disease ◽  
2015 ◽  
Vol 99 (3) ◽  
pp. 422-422 ◽  
Author(s):  
S. Bratsch ◽  
D. Mollov ◽  
B. Lockhart ◽  
D. Johnson ◽  
S. Ehlenbeck
Keyword(s):  
Electron Microscopy ◽  
United States ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Mixed Infection ◽  
The United States ◽  
Type Ii ◽  
Rt Pcr ◽  
Cmv Infection ◽  
Leaf Deformation

Pachysandra terminalis Siebold & Zucc. (Japanese pachysandra, spurge) is widely used as a groundcover. In early 2012, Japanese pachysandra plants from Missouri, which originated in Pennsylvania, showed symptoms of light and dark green mosaic, leaf deformation, concentric ringspots, and stunting. Initial screening of symptomatic leaf tissue by transmission electron microscopy (TEM) using partially purified extracts confirmed the presence of spherical (~28 nm) and bacilliform (18-nm diameter, 35- to 58-nm length) virus particles. Immunosorbent electron microscopy (ISEM) using antisera to a clover isolate of Alfalfa mosaic virus (AMV) (PVAS 92) and to Cucumber mosaic virus (CMV) (ATCC PVAS-30) obtained from the American Type Culture Collection, Manassas, VA, confirmed the presence of AMV and CMV. No other type of virus-like particles were observed by TEM. After 6 months, nearly 20% of the 4,000 pachysandra cuttings exhibited the described symptoms. However, it is possible that more than 20% of the cuttings were infected with both viruses and not yet exhibiting symptoms. Reverse-transcription PCR (RT-PCR) was done using total RNA extracted with a Qiagen RNeasy kit and Ready-To-Go RT-PCR beads (GE Healthcare, UK Limited, UK). The primer pair CMV-1 (5′-GCCGTAAGCTGGATGGACCA) and CMV-2 (5′-TATGATAAGAAGCTTGTTTTCGCG) were used (3) to obtain a 502-bp amplicon from the coat protein (CP) region of CMV RNA 3. The product was ligated and cloned (pGEM-T Easy Vector System; Promega, USA). Three clones were sequenced (UMGC, USA), and the consensus sequence (Sequencher 5.1, Gene Codes Corp., USA) was deposited in GenBank (Accession No. JX227938). The sequence obtained had 100% identity with a homologous CP CMV sequence (AFQ94058) and 99% identity with several other homologous CP CMV sequences (CAX62443, CCK24369, and 15 others). It also contained an EcoRI site at nucleotides 332 to 337, characteristic of CMV Type II isolates (3). The primer pair AMV1F (5′-ATCCACCGATGCCAGCCTTA) and AMV1R (5′-TTCCGCCTCACTGCTGCTG) generated a 1,047-bp product from AMV RNA1 that was deposited in GenBank (JX227937). This product had 100% identity with a homologous AMV sequence (AFQ94057), and 99% identity with several other homologous AMV sequences (AGV15824, ADO85715, CBX36144). From the data presented here, it was concluded that the pachysandra had a mixed infection of AMV and a Type II isolate of CMV. Occurrence of AMV in pachysandra was first reported in New Jersey in 1982 (2) and reported for the first time in France and Germany in 2000 (1). The presence of CMV infection in pachysandra has not been reported in the present literature. Some of the symptoms associated with AMV infection in pachysandra in New Jersey (2) and Europe (1) were similar to the symptoms produced by pachysandra plants infected with both viruses (ring spots, mosaic, and line patterns). However, some symptoms were unique to the mixed infection in pachysandra by AMV and CMV (leaf deformation, stunting). A potential source of this co-infection could occur when plants are grown near alfalfa fields (AMV infection by aphids) and undergo vegetative propagation (CMV infection by contaminated tools). This is the first report of pachysandra co-infected by AMV and CMV in the United States. References: (1) L. Cardin and B. Moury. Plant Dis. 84:594, 2000. (2) D. E. Hershman and E. H. Varney. Plant Dis. 66:1195, 1982. (3) S. Wylie et al. Aust. J. Agric. Res. 44:41, 1993.

scholarly journals First report of dasheen mosaic virus infecting taro (Colocasia esculenta) in Louisiana

Plant Disease ◽  
2021 ◽  
Author(s):  
Cesar Escalante ◽  
David Galo ◽  
Rodrigo Diaz ◽  
Rodrigo Valverde
Keyword(s):  
United States ◽  
Mosaic Virus ◽  
Consensus Sequence ◽  
The United States ◽  
Colocasia Esculenta ◽  
Rt Pcr ◽  
Serological Tests ◽  
First Report ◽  
Pcr Products ◽  
The University

Taro [Colocasia esculenta (L.) Schott], also called dasheen or malanga is an important staple crop in many tropical and subtropical countries (Chaïr et al. 2016). In October 2020, taro plants showing foliar symptoms consisting of mosaic, feathery mottle, and vein clearing patterns were observed in the Hilltop Arboretum, the Bluebonnet Swamp Nature Center, the Louisiana State University Agricultural Center Botanic Gardens, and the University Lake, in Baton Rouge, Louisiana. Unidentified aphids were also observed infesting the plants showing the described symptoms. From each location, two foliar samples from symptomatic and two from asymptomatic plants were collected and tested by ELISA using antiserum for general potyvirus group (Agdia, Elkhart, IN). Seven of eight symptomatic samples tested positive while the asymptomatic samples were negative. The seven positive samples were used to perform an additional ELISA test using antiserum specific for dasheen mosaic virus (DsMV) (Agdia). All seven samples tested positive for DsMV. To confirm the identity of the virus, total RNA was extracted from the seven samples using the PureLink® Plant RNA Reagent Kit (Invitrogen, Carlsbad, CA). After DNA digestion with PerfeCta® DNase I (Qiagen, Beverly, MA), the RNA was used to perform reverse transcription polymerase chain reaction (RT-PCR) with primer set DMV 5708-5731-F/DMV 6131-6154-R which is specific for DsMV (Wang et al. 2017). RT-PCR was performed using the AccessQuickTM RT-PCR System (Promega, Madison, WI) following the reaction conditions described by Wang et al. PCR products of the expected size (~447 bp) were obtained with all seven samples and were Sanger-sequenced. A consensus sequence (MW284936) was obtained with the two sequences from samples collected at the University Lake and aligned with other sequences available in the GenBank using BLASTn. Our isolate of DsMV showed 90.6% nt identity to an isolate of DsMV from Ethiopia (MG602229). Mechanical inoculations to healthy taro plants were conducted using leaf tissue of symptomatic plants as source of inoculum. Inoculated plants exhibited mosaic symptoms three weeks after inoculation and were ELISA-positive for DsMV. Symptomatology, serological tests, RT-PCR testing, and DNA sequencing of RT-PCR products support that the symptomatic taro plants were infected with DsMV. Taro is a crop in Hawaii, but in the contiguous United States, it is mostly grown as an ornamental and is considered an invasive species. Its distribution is restricted to the southern continental states and Hawaii (Cozad et al. 2018). CABI, EPPO (1998) lists the presence of DsMV in several states of the United States, including Louisiana; however, there is no record in the literature of the identification of this virus in Louisiana. The potential impact of DsMV in taro and related ornamental species in southern United States is unknown. To the best of our knowledge, this is the first report documenting DsMV infecting taro in Louisiana.

scholarly journals First Report of Potato spindle tuber viroid Naturally Infecting Field Tomatoes in the Dominican Republic

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 701-701
Author(s):  
K.-S. Ling ◽  
R. Li ◽  
D. Groth-Helms ◽  
F. M. Assis-Filho
Keyword(s):  
United States ◽  
Dominican Republic ◽  
Mosaic Virus ◽  
Potato Spindle Tuber Viroid ◽  
The United States ◽  
Disease Outbreak ◽  
Ringspot Virus ◽  
Rt Pcr ◽  
Spot Virus ◽  
Three Samples

In recent years, viroid disease outbreaks have resulted in serious economic losses to a number of tomato growers in North America (1,2,3). At least three pospiviroids have been identified as the causal agents of tomato disease, including Potato spindle tuber viroid (PSTVd), Tomato chlorotic dwarf viroid (TCDVd), and Mexican papita viroid (MPVd). In the spring of 2013, a severe disease outbreak with virus-like symptoms (chlorosis and plant stunting) was observed in a tomato field located in the Dominican Republic, whose tomato production is generally exported to the United States in the winter months. The transplants were produced in house. The disease has reached an epidemic level with many diseased plants pulled and disposed of accordingly. Three samples collected in May of 2013 were screened by ELISA against 16 common tomato viruses (Alfalfa mosaic virus, Cucumber mosaic virus, Impatiens necrotic spot virus, Pepino mosaic virus, Potato virus X, Potato virus Y, Tobacco etch virus, Tobacco mosaic virus, Tobacco ringspot virus, Tomato aspermy virus, Tomato bushy stunt virus, Tomato mosaic virus, Tomato ringspot virus, Tomato spotted wilt virus, Groundnut ringspot virus, and Tomato chlorotic spot virus), a virus group (Potyvirus group), three bacteria (Clavibacter michiganensis subsp. michiganensis, Pectobacterium atrosepticum, and Xanthomonas spp.), and Phytophthora spp. No positive result was observed, despite the presence of symptoms typical of a viral-like disease. Further analysis by RT-PCR using Agdia's proprietary pospiviroid group-specific primer resulted in positive reactions in all three samples. To determine which species of pospiviroid was present in these tomato samples, full-genomic products of the expected size (~360 bp) were amplified by RT-PCR using specific primers for PSTVd (4) and cloned using TOPO-TA cloning kit (Invitrogen, CA). A total of 8 to 10 clones from each isolate were selected for sequencing. Sequences from each clone were nearly identical and the predominant sequence DR13-01 was deposited in GenBank (Accession No. KF683200). BLASTn searches into the NCBI database demonstrated that isolate DR13-01 shared 97% sequence identity to PSTVd isolates identified in wild Solanum (U51895), cape gooseberry (EU862231), or pepper (AY532803), and 96% identity to the tomato-infecting PSTVd isolate from the United States (JX280944). The relatively lower genome sequence identity (96%) to the tomato-infecting PSTVd isolate in the United States (JX280944) suggests that PSTVd from the Dominican Republic was likely introduced from a different source, although the exact source that resulted in the current disease outbreak remains unknown. It may be the result of an inadvertent introduction of contaminated tomato seed lots or simply from local wild plants. Further investigation is necessary to determine the likely source and route of introduction of PSTVd identified in the current epidemic. Thus, proper control measures could be recommended for disease management. The detection of this viroid disease outbreak in the Dominican Republic represents further geographic expansion of the viroid disease in tomatoes beyond North America. References: (1). K.-S. Ling and M. Bledsoe. Plant Dis. 93:839, 2009. (2) K.-S. Ling and W. Zhang. Plant Dis. 93:1216, 2009. (3) K.-S. Ling et al. Plant Dis. 93:1075, 2009. (4) A. M. Shamloul et al. Can. J. Plant Pathol. 19:89, 1997.

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 Allamanda Mosaic Caused by Cucumber mosaic virus in Taiwan

Plant Disease ◽  
2005 ◽  
Vol 89 (5) ◽  
pp. 529-529 ◽  
Author(s):  
Y. K. Chen ◽  
C. C. Yang ◽  
H. T. Hsu
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Chenopodium Quinoa ◽  
Rabbit Antiserum ◽  
Nucleotide Sequence Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Indicator Plants

Allamanda (Allamanda cathartica L., family Apocynaceae) is native to Brazil and is a popular perennial shrub or vine ornamental in Taiwan. Plants showing severe mosaic, rugosity, and leaf distortion symptoms on leaves are common in commercial nurseries and private gardens. Examination of crude sap prepared from symptomatic leaves using an electron microscope revealed the presence of spherical virus particles with a diameter of approximately 28 nm. The virus was mechanically transmitted to indicator plants and induced symptoms similar to those incited by Cucumber mosaic virus (CMV). The virus caused local lesions on inoculated leaves of Chenopodium quinoa and C. amaranticolor and systemic mosaic in Cucumis sativus, Lycopersicon esculentum, Nicotiana benthamiana, N. glutinosa, N. rustica, and N. tabacum. On N. tabacum, necrotic ringspots developed on inoculated leaves followed by systemic mosaic. Tests of leaf sap extracted from naturally infected allamanda and inoculated indicator plants using enzyme-linked immunosorbent assay were positive to rabbit antiserum prepared to CMV. Viral coat protein on transblots of sodium dodecyl sulfate-polyacrylamide gel electrophoresis reacted with CMV subgroup I specific monoclonal antibodies (2). With primers specific to the 3′-half of RNA 3 (1), amplicons of an expected size (1,115 bp) were obtained in reverse transcription-polymerase chain reaction (RT-PCR) using total RNA extracted from infected allamanda and N. benthamiana. The amplified fragment (EMBL Accession No. AJ871492) was cloned and sequenced. It encompasses the 3′ part of the intergenic region of RNA 3 (158 nt), CP ORF (657 nt), and 3′ NTR (300 nt) showing 91.8–98.9% and 71.4–72.8% identities to those of CMV in subgroups I and II, respectively. Results of MspI-digested restriction fragment length polymorphism patterns of the RT-PCR fragment and the nucleotide sequence analysis indicate that the CMV isolate from allamanda belongs to subgroup IB, which is predominant on the island. To our knowledge, CMV is the only reported virus that infects allamanda and was first detected in Brazil (3), and this is the first report of CMV infection in allamanda plants occurring in Taiwan. References: (1) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (2) H. T. Hsu et al. Phytopathology 90:615, 2000. (3) E. W. Kitajima. Acta. Hortic. 234:451, 1988.

scholarly journals Cucumber Mosaic Virus, Tobacco Streak Virus, and Cucumber Mosaic Virus Satellite RNA Associated with Mosaic and Ringspot Symptoms in Ajuga reptans in Ohio

Plant Disease ◽  
1997 ◽  
Vol 81 (10) ◽  
pp. 1214-1214 ◽  
Author(s):  
J. R. Fisher ◽  
S. T. Nameth
Keyword(s):  
United States ◽  
Molecular Weight ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
The United States ◽  
Negative Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Molecular Weight ◽  
Streak Virus ◽  
Ajuga Reptans

Creeping bugleweed (Ajuga reptans L.) is a perennial ornamental commonly grown as a ground cover in temperate climates. Commercial samples of the A. reptans cultivars Royalty, var. Atropurpurea Bronze, Bronze Beauty, and Burgundy Glow showing mosaic and ringspot symptoms were tested for the presence of virus infection by direct antibody sandwich enzyme-linked immunosorbent assay (ELISA) and viral-associated double-stranded (ds) RNA analysis. Cucumber mosaic cucumovirus (CMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested. ELISA values were considered positive if the absorbance values were twice the negative control. Negative control values were established with asymptomatic tissue of the cv. Bronze Beauty. Tobacco streak ilarvirus (TSV) was detected only by ELISA in symptomatic samples of all cultivars except Royalty. No dsRNA suggestive of TSV was detected. Alfalfa mosaic virus (AMV) was detected by ELISA and dsRNA analysis in symptomatic samples of all cultivars tested except Royalty and var. Atropurpurea Bronze. dsRNA analysis also indicated the presence of a low molecular weight, possible satellite (sat) RNA associated with all symptomatic and asymptomatic Royalty and var. Atropurpurea Bronze plants tested. Northern (RNA) blot analysis with a digoxigenin-labeled full-length clone of the (S) CARNA-5 (-) CMV satRNA (ATCC no. 45124) confirmed that the low molecular weight RNA associated with the Royalty and var. Atropurpurea Bronze cultivars was indeed CMV satRNA. Only AMV has been previously reported in A. reptans in the United States (1). This is the first report of CMV and its satRNA, as well as TSV, in A. reptans in the United States. Reference: (1) W. T. Schroeder and R. Provvidenti. Plant Dis. Rep. 56:285, 1972.

scholarly journals First Report of Tomato Brown Rugose Fruit Virus Infecting Tomato Crop in Saudi Arabia

Plant Disease ◽  
2021 ◽  
Author(s):  
Ahmed Sabra ◽  
Mohammed Ali Al Saleh ◽  
I. M. Alshahwan ◽  
Mahmoud A. Amer
Keyword(s):  
Saudi Arabia ◽  
Mosaic Virus ◽  
Solanum Lycopersicum ◽  
Tomato Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Pcr Products ◽  
Dark Green ◽  
Leaf Symptoms

Tomato (Solanum lycopersicum L.) is the most economically important member of family Solanaceae and cultivated worldwide and one of the most important crops in Saudi Arabia. The aim of this study is screening of the most common viruses in Riyadh region and identified the presence of tomato brown rugose fruit virus (ToBRFV) in Saudi Arabia. In January 2021, unusual fruit and leaf symptoms were observed in several greenhouses cultivating tomatoes commercially in Riyadh Region, Saudi Arabia. Fruit symptoms showed irregular brown spots, deformation, and yellowing spots which render the fruits non-marketable, while the leaf symptoms included mottling, mosaic with dark green wrinkled and narrowing. These plants presented the symptoms similar to those described in other studies (Salem et al., 2015, Luria et al., 2017). A total 45 Symptomatic leaf samples were collected and tested serologically against suspected important tomato viruses including: tomato chlorosis virus, tomato spotted wilt virus, tomato yellow leaf curl virus, tomato chlorotic spot virus, tomato aspermy virus, tomato bushy stunt virus, tomato black ring virus, tomato ringspot virus, tomato mosaic virus, pepino mosaic virus and ToBRFV using Enzyme linked immunosorbent assay (ELISA) test (LOEWE®, Biochemica, Germany), according to the manufacturers' instructions. The obtained results showed that 84.4% (38/45) of symptomatic tomato samples were infected with at least one of the detected viruses. The obtained results showed that 55.5% (25/45) of symptomatic tomato samples were found positive to ToBRFV, three out of 25 samples (12%) were singly infected, however 22 out of 45 (48.8%) had mixed infection between ToBRFV and with at least one of tested viruses. A sample with a single infection of ToBRFV was mechanically inoculated into different host range including: Chenopodium amaranticolor, C. quinoa, C. album, C. glaucum, Nicotiana glutinosa, N. benthamiana, N. tabacum, N. occidentalis, Gomphrena globosa, Datura stramonium, Solanum lycopersicum, S. nigrum, petunia hybrida and symptoms were observed weekly and the systemic presence of the ToBRFV was confirmed by RT-PCR and partial nucleotide sequence. A Total RNA was extracted from DAS-ELISA positive samples using Thermo Scientific GeneJET Plant RNA Purification Mini Kit. Reverse transcription-Polymerase chain reaction (RT-PCR) was carried out using specific primers F-3666 (5´-ATGGTACGAACGGCGGCAG-3´) and R-4718 (5´-CAATCCTTGATGTG TTTAGCAC-3´) which amplified a fragment of 1052 bp of Open Reading Frame (ORF) encoding the RNA-dependent RNA polymerase (RdRp). (Luria et al. 2017). RT-PCR products were analyzed using 1.5 % agarose gel electrophoresis. RT-PCR products were sequenced in both directions by Macrogen Inc. Seoul, South Korea. Partial nucleotide sequences obtained from selected samples were submitted to GenBank and assigned the following accession numbers: MZ130501, MZ130502, and MZ130503. BLAST analysis of Saudi isolates of ToBRFV showed that the sequence shared nucleotide identities ranged between 98.99 % to 99.50 % among them and 98.87-99.87 % identity with ToBRFV isolates from Palestine (MK881101 and MN013187), Turkey (MK888980, MT118666, MN065184, and MT107885), United Kingdom (MN182533), Egypt (MN882030 and MN882031), Jordan (KT383474), USA (MT002973), Mexico (MK273183 and MK273190), Canada (MN549395) and Netherlands (MN882017, MN882018, MN882042, MN882023, MN882024, and MN882045). To our knowledge, this is the first report of occurrence of ToBRFV infecting tomato in Saudi Arabia which suggests its likely introduction by commercial seeds from countries reported this virus and spread in greenhouses through mechanical means. The author(s) declare no conflict of interest. Keywords: Tomato brown rugose fruit virus, tomato, ELISA, RT-PCR, Saudi Arabia References: Luria N, et al., 2017. PLoS ONE 12(1): 1-19. Salem N, et al., 2015. Archives of Virology 161(2): 503-506. Fig. 1. Symptoms caused by ToBRFV showing irregular brown spots, deformation, yellowing spots on fruits (A, B, C) and bubbling and mottling, mosaic with dark green wrinkled and narrowing on leaf (D).

scholarly journals First Report of Blackberry chlorotic ringspot virus in Rubus sp. in the United States

Plant Disease ◽  
2007 ◽  
Vol 91 (4) ◽  
pp. 463-463 ◽  
Author(s):  
I. E. Tzanetakis ◽  
J. D. Postman ◽  
R. R. Martin
Keyword(s):  
United States ◽  
The United States ◽  
Ringspot Virus ◽  
Tobacco Streak Virus ◽  
Rapid Decline ◽  
Streak Virus ◽  
Black Raspberry ◽  
Rt Pcr ◽  
Indicator Plants ◽  
First Report

Blackberry chlorotic ringspot virus (BCRV), genus Ilarvirus, has been found in Rubus sp. in Scotland (2) and rose in the United States (4). The possibility that BCRV infects other hosts in the United States was explored. We tested 18 accessions of Fragaria sp. and 30 of Rubus sp. maintained at the National Clonal Germplasm Repository in Corvallis, OR. Ilarviruses had been detected in these plants by reverse transcription (RT)-PCR, ELISA, or had caused symptoms typical of ilarviruses on indicator plants. The accessions were tested by RT-PCR with primers F (5′-GTTTCCTGTGCTCCTCA-3′) and R (5′-GTCACACCGAGGTACT-3′) (4) that amplify a 519 to 522 nt (depending on the isolate) region of the RNA 3 of BCRV. The virus was detected in two accessions of black raspberry (Rubus occidentalis L.): RUB433, cv. Lowden and RUB 9012, cv. New Logan. The sequences of the fragments amplified from these accessions (GenBank Accession Nos. EF041817 and EF041818, respectively) had 97% nt sequence identity to each other and 95 and 88% nt identity to the rose and Scottish isolates (GenBank Accession Nos. DQ329378 and DQ091195, respectively). Chenopodium quinoa indicator plants inoculated with isolate RUB 433 developed mild chlorotic spots on the inoculated leaves 4 days after inoculation. RT-PCR and sequencing of the amplicons verified BCRV infection of C. quinoa. RUB 9012 was used for the characterization of Black raspberry latent virus (BRLV), later thought to be an isolate of Tobacco streak virus (TSV). This accession was recently found to be infected with Strawberry necrotic shock virus (SNSV) but not TSV (3). It is possible that BRLV may be a mixture of SNSV and BCRV. SNSV is one of the most abundant viruses of Rubus sp. in the Pacific Northwest (1), and the finding of another ilarvirus, BCRV, may account in part for the rapid decline of Rubus sp. observed in several fields in Oregon and Washington. To our knowledge, this is the first report of BCRV infecting Rubus sp. outside the United Kingdom. References: (1) A. B. Halgren. Ph.D. Diss. Oregon State University, Corvallis, OR, 2006. (2) A. T. Jones et al. Ann. Appl. Biol. 149:125, 2006. (3) I. E. Tzanetakis et al. Arch. Virol. 149:2001, 2004. (4) I. E. Tzanetakis et al. Plant Pathol. 55:568, 2006.

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