First report of the infection of alfalfa mosaic virus in Salvia sclarea in Hungary

Viburnum sp. is an ornamental shrub widely used in private and public gardens. It is common in natural wooded areas in the Mediterranean Region. The genus includes more than 150 species distributed widely in climatically mild and subtropical regions of Asia, Europe, North Africa, and the Americas. In January 2007, yellow leaf spotting in young plants of Viburnun lucidum was observed in two ornamental nurseries in the Mediterranean area of Spain. Symptoms appeared sporadically depending on environmental conditions but normally in cooler conditions. Leaf tissue from 24 asymptomatic and five symptomatic plants was sampled and analyzed by double-antibody sandwich (DAS)-ELISA with specific polyclonal antibodies against Tomato spotted wilt virus (TSWV) (Loewe Biochemica, Sauerlach, Germany) and Alfalfa mosaic virus (AMV) (SEDIAG S.A.S, Longvic, France). All symptomatic plants of V. lucidum were positive for Alfalfa mosaic virus (AMV). The presence of AMV was tested in the 29 samples by one-step reverse transcription (RT)-PCR with the platinum Taq kit (Invitrogen Life Technologies, Barcelona, Spain) using primers derived from a partial fragment of the coat protein gene of AMV (2). The RT-PCR assays produced an expected amplicon of 700 bp in the five symptomatic seropositive samples. No amplification product was observed when healthy plants or a water control were used as a template in the RT-PCR assays. One PCR product was purified (High Pure PCR Product Purification Kit; Roche Diagnostics, Mannheim, Germany) and directly sequenced (GenBank Accession No. EF427449). BLAST analysis showed 96% nucleotide sequence identity to an AMV isolate described from Phlox paniculata in the United States (GenBank Accession No. DQ124429). This virosis has been described as affecting Viburnum tinus L. in France (1). To our knowledge, this is the first report of natural infection of Viburnum lucidum with AMV in Spain, which might have important epidemiological consequences since V. lucidum is a vegetatively propagated ornamental plant. References: (1) L. Cardin et al. Plant Dis. 90:1115, 2006. (2) Ll. Martínez-Priego et al. Plant Dis. 88:908, 2004.

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First Report of Alfalfa mosaic virus Infecting Basil (Ocimum basilicum) in California

Plant Disease ◽

10.1094/pdis-06-11-0516 ◽

2012 ◽

Vol 96 (2) ◽

pp. 295-295 ◽

Cited By ~ 3

Author(s):

W. M. Wintermantel ◽

E. T. Natwick

Keyword(s):

Nucleic Acid ◽

Mosaic Virus ◽

Alfalfa Mosaic Virus ◽

Plant Viruses ◽

Ocimum Basilicum ◽

Mechanical Transmission ◽

Rt Pcr ◽

Imperial Valley ◽

First Report ◽

Production Region

Basil (Ocimum basilicum L.) plants collected from three fields in Imperial County, CA in May, 2011 were found to be exhibiting yellowing, chlorotic sectors and spots on leaves, resulting in unmarketable plants. Dodder (Cuscuta spp.) was present in one of the fields, but was not visibly associated with symptomatic plants. Total nucleic acid was extracted from four symptomatic and three asymptomatic basil plants, as well as from the dodder plant with the RNeasy Plant Mini Kit (Qiagen, Valencia, CA). Nucleic acid extracts were tested by reverse transcription (RT)-PCR for the presence of Alfalfa mosaic virus (AMV) using primers designed to amplify a 350-nt region of the AMV coat protein gene (3). RT-PCR produced bands of the expected size in extracts from all symptomatic plants and the dodder sample. No amplification was obtained from symptomless plants. A 350-nt band amplified from one plant was gel-extracted, sequenced (TACGen, Richmond, CA), and confirmed to be AMV by comparison to sequences available in GenBank (Accession No. K02703). Although serological tests on an initial basil sample were negative for AMV by ELISA using antiserum produced against AMV by R. Larsen, USDA-ARS, Prosser, WA (unpublished), AMV was confirmed by ELISA and RT-PCR in symptomatic Nicotiana benthamiana, N. clevelandii, and Malva parviflora plants following mechanical transmission from basil source plants. The fields with AMV infections were located at opposite ends of the production region from one another, indicating widespread dispersal of AMV in the region. All AMV positive plants were adjacent to alfalfa. Two additional basil plantings in shade houses open to the outside environment did not have AMV symptomatic plants and were also confirmed negative by RT-PCR, but these plantings were at the extreme north end of Imperial Valley agriculture and well away from any alfalfa fields. At the time the basil plantations were sampled for AMV, no aphids were found in any plantations, but during the several weeks prior to finding the AMV-positive plants, cowpea aphid, Aphis craccivora Koch; pea aphid, Acyrthosiphon pisum Harris; blue alfalfa aphid, Acyrthosiphon kondoi Shinji; and spotted alfalfa aphid, Therioaphis maculata Buckton were colonizing Imperial Valley alfalfa fields, producing winged adults. AMV is transmitted by at least 14 aphid species (1), and most aphid populations increase during the late spring in this important desert agricultural region. The acquisition of AMV by dodder suggests the parasitic plant may serve as a vector of AMV within basil fields, although further study will be necessary for clarification. Significant acreage of basil is grown in the Imperial Valley. This acreage is surrounded by extensive and increasing alfalfa production totaling 55,442 ha (137,000 acres) in Imperial County and representing a 21% increase in acreage over 2009 for the same region (2). To our knowledge, this is the first report of basil infected by AMV in California. The proximity of basil production to such a large alfalfa production region warrants the need for enhanced efforts at aphid management in basil production to reduce vector populations and reduce transmission to basil crops. References: (1) E. M. Jaspars and L. Bos. Alfalfa mosaic virus. No. 229 in: Descriptions of Plant Viruses. Commonw. Mycol. Inst./Assoc. Appl. Biol., Kew, England, 1980. (2) C. Valenzuela. Imperial County California Crop and Livestock Report, 2010. (3) H. Xu and J. Nie. Phytopathology 96:1237, 2006.

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First Report of Alfalfa mosaic virus in Lavandula officinalis

Plant Disease ◽

10.1094/pdis.2004.88.8.908c ◽

2004 ◽

Vol 88 (8) ◽

pp. 908-908 ◽

Cited By ~ 6

Author(s):

Ll. Martínez-Priego ◽

M. C. Córdoba ◽

C. Jordá

Keyword(s):

Mosaic Virus ◽

Screening Program ◽

Sequence Data ◽

Alfalfa Mosaic Virus ◽

Mediterranean Area ◽

Enzyme Linked Immunosorbent Assay ◽

First Report ◽

Coat Protein Region ◽

C 30 ◽

Lavandula Officinalis

For several years, in ornamental nurseries in the Mediterranean area of Spain, stunting and yellow leaf spotting have been observed in young plants of Lavandula officinalis. Symptoms eventually disappeared as the plants matured. During the summer of 2003, the number of plantlets affected and the intensity of symptoms increased significantly. Symptomatic plants tested positive using enzyme-linked immunosorbent assay (ELISA) (Phyto-Diagnostics, INRA, France) for the presence of Alfalfa mosaic virus (AMV). ELISA results were verified using reverse transcription-polymerase chain reaction (RT-PCR). Total RNA extracts from symptomatic plants were analyzed using primers designed specifically for the coat protein region of AMV utilizing sequence data from GenBank Accession No. AF215664: AMVcoat-F: GT GGT GGG AAA GCT GGT AAA and AMVcoat-R: CAC CCA GTG GAG GTC AGC ATT. The thermocycling schedule was as follows: reverse transcriptase step at 50°C for 30 min, first PCR cycle at 94°C for 2 min, 35 cycles each of 30 s at 94°C, 30 s at 54°C, 30 s at 72°C, followed by a final extension at 72°C for 10 min. A 700-pb PCR product of the expected size was obtained from plants that were positive for AMV using ELISA. The two systems provide for rapid detection of AMV in L. officinalis. A regular screening program will assist in providing virus-free plants to ornamental nurseries. These results demonstrate the presence of AMV in L. officinalis. Alfalfa (Medicago sativa L.) is a typical source of AMV. However, because the nurseries where L. officinalis is grown are not in the vicinity of alfalfa fields, we suggest the source of the infection originated in the propagation material. AMV has currently been reported in L. officinalis only in Italy and France (1). To our knowledge, this is the first report of AMV in L. officinalis in Spain. Reference: (1): A. Garibaldi et al. Ed. Edagricole-Edisioni Agricole della Calderini s.r.l., Bologna, 2000.

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First Report of Alfalfa Mosaic Virus InfectingViburnumin Canada

Plant Disease ◽

10.1094/pd-70-173a ◽

1986 ◽

Vol 70 (2) ◽

pp. 173a ◽

Cited By ~ 1

Author(s):

A. W. Chiko

Keyword(s):

Mosaic Virus ◽

Alfalfa Mosaic Virus ◽

First Report

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First Report of Alfalfa mosaic virus Occurrence in Tecoma capensis in the USA

Plant Health Progress ◽

10.1094/php-2012-0824-02-br ◽

2012 ◽

Vol 13 (1) ◽

pp. 38 ◽

Cited By ~ 1

Author(s):

Benham E. Lockhart ◽

Dimitre Mollov

Keyword(s):

Mosaic Virus ◽

Alfalfa Mosaic Virus ◽

Crop Species ◽

Western Usa ◽

First Report ◽

Landscape Plant ◽

Source Of Infection ◽

The Usa

Tecoma capensis or cape honeysuckle is an evergreen vine-line shrub that is widely used as a landscape plant in many areas of the southern and western USA. This is the first report of Alfalfa mosaic virus (AMV) infection associated with a disease of T. capensis outside of Israel and Spain. In addition to the effect on marketability, AMV infection in cape honeysuckle may be a source of infection of other susceptible crop species since AMV is readily transmitted in a nonpersistent manner by at least 13 species of aphid. Accepted for publication 11 July 2012. Published 24 August 2012.

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First Report of Alfalfa mosaic virus Associated with Severe Mosaic and Mottling of Pepper (Capsicum annuum) and White Clover (Trifolium repens) in Oklahoma

Plant Disease ◽

10.1094/pdis-07-12-0630-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1705-1705 ◽

Cited By ~ 1

Author(s):

O. A. Abdalla ◽

A. Ali

Keyword(s):

United States ◽

Mosaic Virus ◽

Capsicum Annuum ◽

White Clover ◽

Trifolium Repens ◽

Alfalfa Mosaic Virus ◽

The United States ◽

Morphological Data ◽

Total Rna ◽

First Report

Alfalfa mosaic virus (AMV), a member of the genus Alfamovirus, family Bromoviridae (1), has been reported in 44 states in the United States excluding Oklahoma. During a cucurbit survey in the summer of 2010, severe mosaic and mottling symptoms were observed on many peppers (Capsicum annuum) and white clover (Trifolium repens) plants in Tulsa, Oklahoma. Symptomatic leaf samples from 15 pepper and two white clover plants were collected in the Bixby area and analyzed serologically by dot-immunobinding assay (DIBA) using specific polyclonal antibodies against AMV (Agdia, Inc). Seven out of 15 pepper samples and both white clover samples were tested positive by DIBA to AMV. The remaining symptomatic samples were positive to Cucumber mosaic virus (CMV). Total RNA was extracted from DIBA positive AMV samples by Tri-reagent method. A small aliquot of total RNA was tested by reverse transcription (RT)-PCR using specific primers: AMV-F 5′ GTCCGCGATCTCTTAAAT 3′ and AMV-R 5′ GAAGTTTGGGTCGAGAGA 3′ that were designed to amplify 900 bp of the AMV-RNA 3. Analysis of the PCR products on agarose gel electrophoreses showed that all tested samples showed a band of the expected size while DIBA negative AMV samples did not produce any band. The amplified PCR product (900 bp) obtained from pepper and white clover were cleaned with PCR purification kit (Qiagen, Germantown, MD) and directly sequenced bi-directionally using the above primers. Sequence analysis confirmed that this virus shared 97% identity at nucleotide sequence with RNA 3 of AMV isolate from Madison-USA (GenBank Accession No. K02703). For biological and morphological characterization of the virus, eight pepper plants were mechanically inoculated using 0.1 M K2HPO4 buffer (pH 7.2) with total RNA extracted from AMV positive pepper or white clover plant samples. One to two weeks post-inoculation, all inoculated plants produced severe mosaic, mottling, and stunting. Virus-like particles preparations were obtained from these symptomatic plants according to our previously described method (2) and electron microcopy examination showed typical AMV particles. These biological and morphological data further confirmed the presence of AMV infecting pepper and clover in Oklahoma. AMV is a significant pathogen worldwide and infects more than 600 species in 70 families, especially alfalfa, pepper, soybean, and tobacco (3). AMV has a worldwide distribution, including the United States, and particularly the Midwestern U.S. where the incidence of the virus is on the rise recently because of the presence of its vector (Aphis glycines) (4). To our knowledge, this is the first report of AMV infecting crops in Oklahoma, which could pose a threat to other economic crops grown in Oklahoma, especially soybean. References: (1) E. E. Mueller et al. Plant Dis. 91:266, 2007. (2) A. Ali et al. Plant Dis. 96:243, 2012. (3) J. F. Bol. Mol. Plant Path.4:1, 2003. (4) M. Malapi-Nelson et al. Plant Dis.93:1259, 2009.

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First Report of Alfalfa mosaic virus on Safflower in Serbia

Plant Disease ◽

10.1094/pdis-12-14-1267-pdn ◽

2015 ◽

Vol 99 (6) ◽

pp. 896-896 ◽

Cited By ~ 1

Author(s):

D. Milošević ◽

A. Marjanović-Jeromela ◽

D. Jovičić ◽

M. Ignjatov ◽

Z. Nikolić ◽

...

Keyword(s):

Mosaic Virus ◽

Alfalfa Mosaic Virus ◽

First Report

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First Report of Alfalfa mosaic virus Infecting Tedera (Bituminaria bituminosa (L.) C.H. Stirton var. albomarginata and crassiuscula) in Australia

Plant Disease ◽

10.1094/pdis-04-12-0378-pdn ◽

2012 ◽

Vol 96 (9) ◽

pp. 1384-1384 ◽

Cited By ~ 5

Author(s):

R. A. C. Jones ◽

D. Real ◽

S. J. Vincent ◽

B. E. Gajda ◽

B. A. Coutts

Keyword(s):

Mosaic Virus ◽

Alfalfa Mosaic Virus ◽

Plant Viruses ◽

Yellow Mosaic Virus ◽

Rt Pcr ◽

First Report ◽

Pasture Legumes ◽

Bituminaria Bituminosa ◽

Pcr Products ◽

Burr Medic

Tedera (Bituminaria bituminosa (L.) C.H. Stirton vars albomarginata and crassiuscula) is being established as a perennial pasture legume in southwest Australia because of its drought tolerance and ability to persist well during the dry summer and autumn period. Calico (bright yellow mosaic) leaf symptoms occurred on occasional tedera plants growing in genetic evaluation plots containing spaced plants at Newdegate in 2007 and Buntine in 2010. Alfalfa mosaic virus (AlMV) infection was suspected as it often causes calico in infected plants (1,2) and infects perennial pasture legumes in local pastures (1,3). Because AlMV frequently infects Medicago sativa (alfalfa) in Australia and its seed stocks are commonly infected (1,3), M. sativa buffer rows were likely sources for spread by aphids to healthy tedera plants. When leaf samples from plants with typical calico symptoms from Newdegate (2007) and Buntine (2010) were tested by ELISA using poyclonal antisera to AlMV, Bean yellow mosaic virus (BYMV) and Cucumber mosaic virus (CMV), only AlMV was detected. When leaf samples from 864 asymptomatic spaced plants belonging to 34 tedera accessions growing at Newdegate and Mount Barker in 2010 were tested by ELISA, no AlMV, BYMV, or CMV were detected, despite presence of M. sativa buffer rows. A culture of AlMV isolate EW was maintained by serial planting of infected seed of M. polymorpha L. (burr medic) and selecting seed-infected seedlings (1,3). Ten plants each of 61 accessions from the local tedera breeding program were grown at 20°C in an insect-proof air conditioned glasshouse. They were inoculated by rubbing leaves with infective sap containing AlMV-EW or healthy sap (five plants each) using Celite abrasive. Inoculations were always done two to three times to the same plants. When both inoculated and tip leaf samples from each plant were tested by ELISA, AlMV was detected in 52 of 305 AlMV-inoculated plants belonging to 36 of 61 accessions. Inoculated leaves developed local necrotic or chlorotic spots or blotches, or symptomless infection. Systemic invasion was detected in 20 plants from 12 accessions. Koch's postulates were fulfilled in 12 plants from nine accessions (1 to 2 of 5 plants each), obvious calico symptoms developing in uninoculated leaves, and AlMV being detected in symptomatic samples by ELISA, inoculation of sap to diagnostic indicator hosts (2) and RT-PCR with AlMV CP gene primers. Direct RT-PCR products were sequenced and lodged in GenBank. When complete nucleotide CP sequences (666 nt) of two isolates from symptomatic tedera samples and two from alfalfa (Aq-JX112758, Hu-JX112759) were compared with that of AlMV-EW, those from tedera and EW were identical (JX112757) but had 99.1 to 99.2% identities to the alfalfa isolates. JX112757 had 99.4% identity with Italian tomato isolate Y09110. Systemically infected tedera foliage sometimes also developed vein clearing, mosaic, necrotic spotting, leaf deformation, leaf downcurling, or chlorosis. Later-formed leaves sometimes recovered, but plant growth was often stunted. No infection was detected in the 305 plants inoculated with healthy sap. To our knowledge, this is the first report of AlMV infecting tedera in Australia or elsewhere. References: (1) B. A. Coutts and R. A. C. Jones. Ann. Appl. Biol. 140:37, 2002. (2) E. M. J. Jaspars and L. Bos. Association of Applied Biologists, Descriptions of Plant Viruses No. 229, 1980. (3) R. A. C. Jones. Aust. J. Agric. Res. 55:757, 2004.

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First Report of the Natural Infection of Robinia pseudoacacia with Alfalfa mosaic virus

Plant Disease ◽

10.1094/pdis-12-12-1136-pdn ◽

2013 ◽

Vol 97 (6) ◽

pp. 851-851 ◽

Cited By ~ 2

Author(s):

G. Delibašić ◽

B. Tanović ◽

J. Hrustic ◽

I. Stanković ◽

A. Bulajić ◽

...

Keyword(s):

Mosaic Virus ◽

Black Locust ◽

Natural Infection ◽

Robinia Pseudoacacia ◽

Alfalfa Mosaic Virus ◽

Natural Host ◽

Post Inoculation ◽

Rt Pcr ◽

First Report ◽

Negative Controls

Robinia pseudoacacia L. (family Fabaceae), commonly known as black locust, is native to the southeastern United States, but has been widely planted and naturalized in temperate regions worldwide. In Europe it is often planted alongside streets and in parks, not only because of the dense canopy and impressive flower clusters in spring, but also because it tolerates air pollution well. In June 2012, several black locust trees exhibiting yellow leaf spots accompanied by mottling and leaf deformation were observed in a park in Backa Topola, North Backa District, Serbia. Numerous aphid colonies were found colonizing symptomatic trees. Leaves collected from nine symptomatic and 10 asymptomatic trees were tested for the presence of three common aphid-transmitted viruses, Alfalfa mosaic virus (AMV), Cucumber mosaic virus, and Potato virus Y, using double-antibody sandwich (DAS)-ELISA with commercial polyclonal antibody (Bioreba AG, Reinach, Switzerland). Commercial positive and negative controls and extracts from healthy black locust leaves were included in each assay. AMV was serologically detected in all symptomatic and also in four of the asymptomatic trees, while no other tested viruses were found. Sap from affected leaves of a ELISA-positive sample (373-12) was mechanically inoculated onto five plants each of Chenopodium quinoa and Nicotiana benthamiana using 0.01 M phosphate buffer (pH 7). Symptoms including local chlorotic leaf lesions followed by mosaic on C. quinoa and a bright yellow mosaic on N. benthamiana were observed on all inoculated plants 5 and 10 days post-inoculation, respectively. The identity of the virus was confirmed using reverse transcription (RT)-PCR analysis. Total RNAs from all naturally and mechanically infected plants were isolated using RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). RT-PCR was carried out using the One-Step RT-PCR Kit (Qiagen) with primer pair CP AMV1 and CP AMV2 specific to the partial CP gene and 3′-UTR of AMV RNA 3 (1). Total RNAs from Serbian AMV isolate from alfalfa (GenBank Accession No. FJ527748) and RNA extract from healthy leaves of R. pseudoacacia were used as positive and negative controls, respectively. All tested plants, as well as the positive control, yielded an amplicon of the correct predicted size (751 bp), while no amplicon was recorded in the healthy control. The amplified product of isolate 373-12 was purified with QIAquick PCR Purification Kit (Qiagen) and sequenced on ABI PRISM 3700 DNA analyzer (Macrogen, South Korea) in both directions (KC288155). Pairwise comparison of the 373-12 isolate CP sequence with those available in GenBank, conducted with MEGA5 software (4), revealed the maximum nucleotide identity of 99% (99% amino acid identity) with the soybean isolate (HQ185569) from Tennessee. AMV has a worldwide distribution and its natural host range includes over 150 plant species, including many herbaceous and several woody plants (2). To our knowledge, this is the first report of R. pseudoacacia as a natural host of AMV worldwide. This finding has potentially significant implications for the successful production of susceptible crops, considering that black locust could act as an important link in the epidemiology of AMV as it may serve as a virus reservoir (3). References: (1) M. M. Finetti-Sialer et al. J. Plant Pathol. 79:115, 1997. (2) R. Hull. Comparative Plant Virology. 2nd ed. Elsevier Academic Press, Burlington, MA, 2009. (3) E. E. Muller et al. Plant Dis. 96:506, 2012. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

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