First Report of Cucumber mosaic virus in Paeonia lactifera in France

Chinese peony (Paeonia lactiflora Pall.), a hardy ornamental plant of the family Paeoniaceae cultivated in gardens and for cut flower production, is frequently infected by Tobacco rattle virus (TRV) in the field. The virus usually induces severe mosaic and chlorotic ringspot symptoms in the leaves, decreasing the commercial value of cut flowers. TRV is routinely detected by mechanical inoculation onto Nicotiana tabacum cv Xanthi, where it induces typical necrotic local ringspots in 3 to 7 days, followed by a reverse transcription (RT)-PCR test (2). In 2004, Xanthi test plants inoculated with sap extracts from 4 of 36 P. lactiflora cv. Odile plants grown in a field plot in the region of Hyères (southeast France) showed systemic mosaic symptoms in addition to the TRV-typical response. In each case, Cucumber mosaic virus (CMV) was detected by the reactions of a range of inoculated plants (1), the observation of 30 nm isometric particles in crude leaf extracts with the electron microscope, and by positive reactions in double antibody sandwich (DAS)-ELISAs with specific polyclonal antibodies. In double-immunodiffusion analysis, these isolates were shown to belong to the group II of CMV isolates (3). ELISA of the peony plants confirmed the presence of CMV and revealed two additional infected plants in the spring of 2006. Following isolation from local lesions on Vigna unguiculata and multiplication in Xanthi tobacco plants, one of the isolates was used to inoculate manually or with Myzus persicae aphids 10 CMV-free plants of P. lactiflora cv. Odile obtained from meristem culture. Three months postinoculation, only three of the aphid-inoculated plants were CMV positive by DAS-ELISA. No change was observed at 1 year postinoculation and no symptoms have been observed, even in CMV-infected plants. CMV appears to be latent in P. lactiflora, therefore detection of CMV before vegetative propagation of the plants is advised because of the risks of synergism for symptoms with other viruses such as TRV. To our knowledge this is the first report of CMV in peony. References: (1) L. Cardin et al. Plant Dis. 87:1263, 2003. (2) D. J. Robinson J. Virol. Methods 40:55, 1992. (3) M. J. Roossinck. J. Virol. 76:3382, 2002.

Download Full-text

Mosaic Symptoms Induced by Cucumber mosaic virus in Polygala myrtifolia in France and New Zealand

Plant Disease ◽

10.1094/pd-89-0527a ◽

2005 ◽

Vol 89 (5) ◽

pp. 527-527 ◽

Cited By ~ 4

Author(s):

L. Cardin ◽

J.-P. Onesto ◽

B. Delecolle ◽

B. Moury

Keyword(s):

New Zealand ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Polyclonal Antibodies ◽

Virus Titer ◽

Cut Flowers ◽

Total Recovery ◽

Voucher Specimen ◽

Polygala Myrtifolia ◽

Das Elisa

Myrtle-leaf milkwort or sweet pea shrub (Polygala myrtifolia L.), family Polygalaceae, is a shrub from South Africa and is well adapted to Mediterranean-type conditions and used as an ornamental plant in gardens and pots or as cut flowers. During 2002 and 2003, mosaic symptoms and leaf distortion were observed in P. myrtifolia in Menton, Roquebrune-Cap Martin, Golfe Juan, and Antibes (Alpes Maritimes Department, France) in public gardens and potted plants. Occasionally, white streaks were observed in flowers. Cucumber mosaic virus (CMV) was identified in samples collected from the four locations on the basis of transmission to and symptoms exhibited by a range of diagnostic host plants (1), observation of isometric particles (≅30 nm) in crude sap preparations from the infected plants by electron microscopy, and positive reaction using double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) with polyclonal antibodies raised against CMV (2). Each isolate was shown to be a group II CMV strain (3) using double-immunodiffusion analysis. During 2004, CMV was also detected using DAS-ELISA in P. myrtifolia samples collected in New Zealand (Christchurch, Akaroa, and Roturoa). To confirm that CMV was responsible for pathogenicity, the Menton isolate was isolated from local lesions on Vigna unguiculata, amplified in Nicotiana tabacum cv. Xanthi-nc, and then mechanically inoculated into 1-year-old P. myrtifolia, P. myrtifolia cv. Grandiflora, and P. myrtifolia cv. Compacta (synonymous to cv. Nana) plants. The D strain of CMV, a reference tomato strain from subgroup I (2), was used for comparison. All experimental plants were propagated from cuttings, grown hydroponically and all tested negative for CMV using DAS-ELISA prior to inoculation. At 12 weeks postinoculation, systemic symptoms were observed on leaves from all inoculated plants (10 plants per genotype for the Menton isolate and 5 plants per genotype for the D strain), except for two P. myrtifolia plants inoculated with the Menton isolate. CMV was detected in apical, noninoculated leaves using DAS-ELISA in all symptomatic plants. A total recovery from symptoms was observed in P. myrtifolia and P. myrtifolia cv. Grandiflora but not in P. myrtifolia cv. Compacta at 6 months postinoculation (mpi) in 7 of 15, 10 of 15, and 15 of 15 DAS-ELISA positive plants, respectively. At 7 mpi, the plants were pruned and planted in soil and at 8 mpi, CMV was detected using DAS-ELISA in most of the plants, and symptoms developed in a few stems of some of the plants. Tessitori et al. (4) described similar symptoms and have detected CMV in P. myrtifolia from Italy, but they did not reproduce the disease in healthy plants. Our results show that CMV is responsible for the symptoms observed and that both CMV subgroups are infectious in P. myrtifolia. Since P. myrtifolia is generally vegetatively propagated by cuttings, frequent CMV tests on the mother stock plants are recommended because of fluctuations in virus titer and symptom expression in some genotypes. To our knowledge, this is the first report of this CMV host in France and New Zealand. A voucher specimen will be deposited at the Station de Pathologie Végétale at INRA, Montfavet. References: (1) L. Cardin et al. Plant Dis. 87:1263, 2003. (2) J. C. Devergne and L. Cardin. Ann. Phytopathol. 7:225, 1975. (3) M. J. Roossinck. J. Virol. 76:3382, 2002. (4) M. Tessitori et al. Plant Dis. 86:1403, 2002.

Download Full-text

First Report of Cucumber mosaic virus in Echium candicans in France

Plant Disease ◽

10.1094/pdis-91-11-1516c ◽

2007 ◽

Vol 91 (11) ◽

pp. 1516-1516 ◽

Cited By ~ 2

Author(s):

L. Cardin ◽

B. Moury

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Polyclonal Antibodies ◽

Chenopodium Quinoa ◽

First Report ◽

Deep Blue ◽

Subgroup Ii ◽

Perennial Shrub ◽

Young Leaves ◽

Das Elisa

Echium candicans (Linn.) Herb. Banks (Pride of Madeira or Viper's Bugloss), family Boraginaceae, is a perennial shrub used in gardens for the ornamental quality of its deep blue inflorescences, especially in coastal areas near the Mediterranean Sea. Mosaic symptoms were observed in leaves of E. candicans in the Alpes Maritimes Department of southeastern France, St Jean Cap Ferrat in 1994, Menton in 2002, and Antibes in 2005. Symptoms exhibited in a range of inoculated plants including Nicotiana tabacum cvs. Xanthi and Samsun, Chenopodium quinoa, C. amaranticolor, Vigna unguiculata cv. Black, and Cucumis sativus cv. Poinsett were typical of Cucumber mosaic virus (CMV). Occurrence of CMV in one sample from each of the three localities was confirmed by the observation of isometric particles (approximately 30 nm) with the electron microscope in crude sap preparations from the infected plants, positive reactions in double-antibody sandwich (DAS)-ELISA to polyclonal antibodies raised against CMV (1), and the nonpersistent transmission of the virus from infected Xanthi to virus-free Xanthi plants by Myzus persicae. In double-immunodiffusion analysis, the three isolates were shown to belong to the CMV subgroup II (1,2). To determine if CMV was responsible for the symptoms observed, the isolate from Antibes was multiplied in Xanthi plants after isolation from local lesions on V. unguiculata and mechanically inoculated to 3-year-old plants of E. candicans tested to be free from CMV before the mechanical inoculation. One month after inoculation, mild mosaic symptoms were observed in young leaves and CMV was detected by DAS-ELISA in 10 of 10 inoculated plants. To our knowledge, this is the first report of CMV in E. candicans. References: (1) J.-C. Devergne and L. Cardin. Ann. Phytopathol. 7:225, 1975. (2) M. J. Roossinck. J. Virol. 76:3382, 2002.

Download Full-text

First Report of Tomato torrado virus Infecting Tomato in Colombia

Plant Disease ◽

10.1094/pdis-11-11-1000 ◽

2012 ◽

Vol 96 (4) ◽

pp. 592-592 ◽

Cited By ~ 9

Author(s):

M. Verbeek ◽

A. M. Dullemans

Keyword(s):

Mosaic Virus ◽

Potato Virus ◽

Polyclonal Antibodies ◽

Sandwich Elisa ◽

Potato Virus X ◽

Rt Pcr ◽

Leaf Extracts ◽

Serological Tests ◽

First Report ◽

Initial Symptoms

Tomato (Solanum lycopersicum L.) plants grown in plastic greenhouses near Villa de Leyva, northeast of Bogota, Colombia showed necrotic spots on the leaves in September 2008. Initial symptoms were necrosis beginning at the base of leaflets that were surrounded by yellow areas. These symptoms resembled those described for Tomato torrado virus (ToTV; family Secoviridae, genus Torradovirus), which was first found in Spain (2). Other (tentative) members of the genus Torradovirus, Tomato marchitez virus (ToMarV), Tomato chocolate spot virus (ToChSV), and Tomato chocolàte virus (ToChV) (3) induce similar symptoms on tomato plants. One sample, coded T418, was stored in the freezer and brought to our lab in 2011. Serological tests (double-antibody sandwich-ELISA) using polyclonal antibodies (Prime Diagnostics, Wageningen, The Netherlands) on leaf extracts showed the absence of Pepino mosaic virus (PepMV), Tobacco mosaic virus (TMV), Tomato spotted wilt virus (TSWV), Cucumber mosaic virus (CMV), Potato virus X (PVX), and Potato virus Y (PVY). Leaf extracts were mechanically inoculated onto the indicator plants Physalis floridana, Nicotiana hesperis ‘67A’, and N. occidentalis ‘P1’ (six plants in total) and were kept in a greenhouse at 20°C with 16 h of light. Necrotic symptoms appeared 4 to 5 days postinoculation and resembled those described for ToTV (2). Two dip preparations of systemically infected P. floridana and N. occidentalis leaves were examined by electron microscopy, which revealed the presence of spherical virus particles of approximately 30 nm. To confirm the presence of ToTV, total RNA was extracted from the original leaf material and an inoculated P. floridana and N. occidentalis plant using the Qiagen Plant Mini Kit (Qiagen, Hilden, Germany) following manufacturer's instructions. ToTV-specific primer sets ToTV-Dp33F/ToTV-Dp20R (5′-TGCTCAATGTTGGAAACCCC-3′/5′-AGCCCTTCATAGGCTAGCC-3′, amplifying a fragment of the RNA1 polyprotein with an expected size of 751 bp) and ToTV-Dp1F/ToTV-Dp2R (5′-ACAAGAGGAGCTTGACGAGG-3′/5′-AAAGGTAGTGTAATGGTCGG-3′, amplifying a fragment on the RNA2 movement protein region with an expected size of 568 bp) were used to amplify the indicated regions in a reverse transcription (RT)-PCR using the One-Step Access RT-PCR system (Promega, Madison, WI). Amplicons of the predicted size were obtained in all tested materials. The PCR products were purified with the Qiaquick PCR Purification Kit (Qiagen) and sequenced directly. BLAST analyses of the obtained sequences (GenBank Accession Nos. JQ314230 and JQ314229) confirmed the identity of isolate T418 as ToTV, with 99% identity to isolate PRI-ToTV0301 in both fragments (GenBank Accession Nos. DQ388879 and DQ388880 for RNA1 and RNA 2, respectively). To our knowledge, this is the first report of ToTV in Colombia, and interestingly, since ToTV has been found only in Europe and Australia (1) so far, this is the first report of ToTV on the American continent. References: (1) C. F. Gambley et al. Plant Dis. 94:486, 2010. (2) M. Verbeek et al. Arch. Virol. 152:881, 2007. (3) M. Verbeek et al. Arch. Virol. 155:751, 2010.

Download Full-text

First Report of Cucumber mosaic virus in Helleborus foetidus in France and Italy

Plant Disease ◽

10.1094/pdis.2003.87.10.1263b ◽

2003 ◽

Vol 87 (10) ◽

pp. 1263-1263 ◽

Cited By ~ 2

Author(s):

L. Cardin ◽

J. P. Onesto ◽

B. Moury

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Western Europe ◽

Chenopodium Quinoa ◽

Mechanical Transmission ◽

White Oak ◽

First Report ◽

Helleborus Foetidus ◽

Group I ◽

Das Elisa

Helleborus foetidus L. (bear's foot) is a perennial plant from the family Ranunculaceae that is common in chalky soils of southern and western Europe. It is grown in gardens for its palm-shaped leaves and early flowers. In 1995, yellow-to-white oak leaf and line patterns in leaves of H. foetidus plants were observed in Hunawihr (Alsace, France). The same symptoms were observed in plants in Entrevaux, Biot, and Gourdon (Provence-Alpes-Côte d'Azur, France) in 2000 and 2001, in Triora (Liguria, Italy) in 2002, and on cv. Western Flisk in a nursery in Nice (Provence-Alpes-Côte d'Azur, France) in 2002. Samples collected from these six locations contained six isolates that were further characterized. Sap extracted from symptomatic plants was mechanically inoculated onto Nicotiana tabacum cvs. Xanthi-nc and Samsun, Chenopodium quinoa, C. amaranticolor, Vigna unguiculata cv. Black, and Cucumis sativus cv. Poinsett. Symptoms exhibited by the inoculated plants indicated infection by Cucumber mosaic virus (CMV). Sap extracted from symptomatic plants reacted positively in double-antibody sandwich-enzyme-linked immunosorbent assays (DAS-ELISA) to antibodies raised against CMV (2). Isometric particles (approximately 30 nm) were observed with an electron microscope in crude sap preparations from infected plants. Following purification of the suspect virus from infected N. tabacum (2) and treatment with formaldehyde (1), each isolate was shown to belong to group II of CMV strains (1,3) by double-immunodiffusion analysis. Following isolation from local lesions on V. unguiculata, the Hunawihr isolate was grown in cv. Xanthi-nc plants and back-inoculated to 2-year-old uninfected seedlings of H. foetidus by aphids (Myzus persicae) or mechanical transmission. Mechanical transmissions were also performed with sap extracted from cv. Xanthi-nc plants infected with the D strain, which belongs to group I of CMV strains (3). Three months postinoculation, symptoms previously described in the original plants were observed in 3 of 10 mechanically inoculated plants and in 2 of 14 aphid-inoculated plants (Hunawihr isolate), whereas no symptoms could be seen in any of the six plants inoculated with the D strain. On the basis of DAS-ELISA, 7 of 10 plants mechanically inoculated and 7 of 14 plants aphid inoculated with the Hunawihr isolate were infected with CMV, whereas 3 of the 6 plants inoculated with the D strain were infected with CMV. To our knowledge, this is the first report that H. foetidus is a natural host for CMV. Beyond the direct impact of the disease induced by CMV on H. foetidus, this perennial and widespread plant species can be an important reservoir of CMV. References: (1) J. C. Devergne and L. Cardin. Ann. Phytopathol. 7:225, 1975. (2) J. C. Devergne et al. Ann. Phytopathol. 10:233, 1978. (3) M. J. Roossinck. J. Virol. 76:3382, 2002.

Download Full-text

First Report of Tobacco rattle virus and Cucumber mosaic virus in Phlox paniculata in France

Plant Disease ◽

10.1094/pdis-91-3-0322b ◽

2007 ◽

Vol 91 (3) ◽

pp. 322-322 ◽

Cited By ~ 1

Author(s):

L. Cardin ◽

J. P. Onesto ◽

I. Bornard ◽

B. Moury

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Tobacco Rattle Virus ◽

Post Inoculation ◽

Mild Mosaic ◽

Rt Pcr ◽

Chenopodium Amaranticolor ◽

Phlox Paniculata ◽

Local Lesions ◽

Das Elisa

Phlox paniculata L., a perennial plant from the family Polemoniaceae, is cultivated as an ornamental in gardens and for cut-flower production. In spring 2003, two types of symptoms were observed in P. paniculata plants grown for cut flowers on a farm in the Var department, France. Some plants showed a mild leaf mosaic while others showed leaf browning and delayed growth. In plants showing mild mosaic, Cucumber mosaic virus (CMV) was detected on the basis of the symptoms exhibited by a range of inoculated plants, the observation of isometric particles (approximately 30 nm) with the electron microscope in crude sap preparations from the infected plants, and the positive reaction in double-antibody sandwich (DAS)-ELISA to polyclonal antibodies raised against CMV (1). In double-immunodiffusion analysis, the five tested isolates were shown to belong to group II of CMV strains. To determine if CMV was responsible for the symptoms observed, one isolate was multiplied in Nicotiana tabacum cv. Xanthi-nc plants after isolation from local lesions on Vigna unguiculata and mechanically inoculated to 12 1-year-old P. paniculata plants. At 3 months post inoculation (mpi), all plants showed mild mosaic and CMV was detected by DAS-ELISA. In sap preparations from P. paniculata plants showing leaf browning symptoms, rod-shaped particles with two distinct sizes of 190 to 210 and 70 to 90 nm long, typical of those associated with tobraviruses, were revealed using electron microscopy. Local lesions typical of Tobacco rattle virus (TRV) were observed after inoculation of N. tabacum cv. Xanthi-nc, Chenopodium amaranticolor, and C. quinoa. Total nucleic acid preparations were prepared from symptomatic plants, and amplicons of the expected size (463 bp) were generated by reverse-transcription (RT)-PCR using primers specific to TRV RNA 1 (4). The nucleotide sequence of one amplicon was 93.6% identical to the sequence of a reference TRV isolate (GenBank Accession No. AJ586803). Twelve 1-year-old P. paniculata plants were mechanically inoculated with an extract of infected tissues from one symptomatic P. paniculata plant. TRV was detected 2 to 6 mpi in apical leaves of all inoculated plants by RT-PCR, although the plants did not express symptoms. Since no other pathogens were detected in the source plants, it is plausible that the lack of symptoms in back-inoculated plants is either due to a long incubation period or an interaction with particular environmental factors such as cold conditions. The survey of approximately 200 plants revealed that approximately 7, 10, and 1% were infected by TRV, CMV, or by both viruses, respectively. CMV and TRV were previously detected in P. paniculata in Latvian SSR and in Lithuania (2,3). These results show that sanitary selection of P. paniculata prior to vegetative propagation should include a screening for TRV and CMV infections. References: (1) J.-C. Devergne et al. Ann. Phytopathol. 10:233, 1978. (2) Y. Ignab and A. Putnaergle. Tr. Latv. S.-Kh. Akad. 118:27, 1977. (3) M. Navalinskiene and M. Samuitiene. Biologija 1:52, 1996. (4) D. J. Robinson. J. Virol. Methods 40:57, 1992.

Download Full-text

Natural occurrence of Cucumber mosaic virus infecting water mint (Mentha aquatica) in Antalya and Konya, Turkey

Acta Botanica Croatica ◽

10.2478/v10184-011-0052-7 ◽

2012 ◽

Vol 71 (1) ◽

pp. 187-193 ◽

Cited By ~ 4

Author(s):

Mehmet Sevik

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Mediterranean Region ◽

Mountain Range ◽

Natural Occurrence ◽

First Report ◽

Mentha Aquatica ◽

Taurus Mountains ◽

Mint Leaf ◽

Das Elisa

Natural occurrence ofCucumber mosaic virusinfecting water mint (Mentha aquatica) in Antalya and Konya, TurkeyAvirus causing a disease in mint (the aromatic and culinary plant) has recently become a problem in the Taurus Mountains, a mountain range in the Mediterranean region of Turkey. To detect the virus and investigate its distribution in the region, mint leaf samples were collected from the vicinity of spring areas in the plateaus of Antalya and Konya in 2009. It was found that Cucumber mosaic virus (CMV) was detected in 27.08% of symptomatic samples tested by DAS-ELISA. To the best of our knowledge, this is the first report of CMV on mint plants in this region of Turkey.

Download Full-text

First Report of Cucumber mosaic virus on Melon in Bosnia and Herzegovina

Plant Disease ◽

10.1094/pdis-02-13-0135-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1124-1124 ◽

Cited By ~ 1

Author(s):

V. Trkulja ◽

D. Kovačić ◽

B. Ćurković ◽

A. Vučurović, I. Stanković ◽

A. Bulajić ◽

...

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Bosnia And Herzegovina ◽

Cucumis Melo ◽

Cucurbita Pepo ◽

Yellow Mosaic Virus ◽

Rt Pcr ◽

First Report ◽

Negative Controls ◽

Das Elisa

During July 2012, field-grown melon plants (Cucumis melo L.) with symptoms of mosaic, chlorotic mottling, and vein banding as well as blistering and leaf malformation were observed in one field in the locality of Kladari (municipality of Doboj, Bosnia and Herzegovina). Disease incidence was estimated at 60%. A total of 20 symptomatic plants were collected and tested with double-antibody sandwich (DAS)-ELISA using commercial polyclonal antisera (Bioreba AG, Reinach, Switzerland) against four the most commonly reported melon viruses: Cucumber mosaic virus (CMV), Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), and Papaya ringspot virus (PRSV) (1,3). Commercial positive and negative controls were included in each assay. Only CMV was detected serologically in all screened melon samples. Sap from an ELISA-positive sample (162-12) was mechanically inoculated to test plants using 0.01 M phosphate buffer (pH 7.0). The virus caused necrotic local lesions on Chenopodium amaranticolor 5 days after inoculation, while mild to severe mosaic was observed on Nicotiana rustica, N. glutinosa, N. tabacum ‘Samsun,’ Cucurbita pepo ‘Ezra F1,’ and Cucumis melo ‘Ananas’ 10 to 14 days post-inoculation. All five inoculated plants of each experimental host were DAS-ELISA positive for CMV. The presence of CMV in all naturally and mechanically infected plants was further verified by conventional reverse transcription (RT)-PCR. Total RNAs were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions and used as template in RT-PCR. RT-PCR was carried out with the One-Step RT-PCR Kit (Qiagen) using primer pair CMVCPfwd and CMVCPrev (4), amplifying the entire coat protein (CP) gene and part of 3′- and 5′-UTRs of CMV RNA 3. Total RNAs obtained from the Serbian CMV isolate from Cucurbita pepo ‘Olinka’ (GenBank Accession No. HM065510) and healthy melon leaves were used as positive and negative controls, respectively. An amplicon of the correct predicted size (871 bp) was obtained from all naturally and mechanically infected plants as well as from positive control, but not from healthy tissues. The amplified product derived from isolate 162-12 was purified with QIAquick PCR Purification Kit (Qiagen) and sequenced directly using the same primer pair as in RT-PCR (KC559757). Multiple sequence alignment of the 162-12 isolate CP sequence with those available in GenBank, conducted with MEGA5 software, revealed that melon isolate from Bosnia and Herzegovina showed the highest nucleotide identity of 99.7% (100% amino acid identity) with eight CMV isolates originating from various hosts from Serbia (GQ340670), Spain (AJ829770 and 76, AM183119), the United States (U20668, D10538), Australia (U22821), and France (X16386). Despite the fact that CMV is well established in majority of Mediterranean countries and represents an important threat for many agriculture crops, including pepper in Bosnia and Herzegovina (2), to our knowledge, this is the first report of CMV infecting melon in Bosnia and Herzegovina. Melon popularity as well as production value has been rising rapidly and the presence of CMV may have a drastic economic impact on production of this crop in Bosnia and Herzegovina. References: (1) E. E. Grafton-Cardwell et al. Plant Dis. 80:1092, 1996. (2) M. Jacquemond. Adv. Virus Res. 84:439, 2012. (3) M. Luis-Arteaga et al. Plant Dis. 82:979, 1998. (4) K. Milojević et al. Plant Dis. 96:1706, 2012.

Download Full-text

First Report of Cucumber mosaic virus Infecting Peperomia tuisana in Serbia

Plant Disease ◽

10.1094/pdis-01-13-0089-pdn ◽

2013 ◽

Vol 97 (7) ◽

pp. 1004-1004 ◽

Cited By ~ 2

Author(s):

K. Milojević ◽

I. Stanković ◽

A. Vučurović ◽

D. Ristić ◽

D. Milošević ◽

...

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Disease Incidence ◽

Impatiens Necrotic Spot Virus ◽

Rt Pcr ◽

Test Species ◽

First Report ◽

Cp Gene ◽

Negative Controls ◽

Das Elisa

Peperomia tuisana C.DC. ex Pittier (family Piperaceae) is an attractive succulent grown as an ornamental. Despite its tropical origins, it can be successfully grown indoors in any climate. In March 2012, three samples of P. tuisana showing virus-like symptoms were collected from a commercial greenhouse in Zemun (District of Belgrade, Serbia) in which estimated disease incidence was 80%. Infected plants showed symptoms including necrotic ringspots and line patterns that enlarged and caused necrosis of leaves. A serious leaf drop led to growth reduction and even death of the plant. Leaves from three symptomatic P. tuisana plants were sampled and analyzed by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kits (Bioreba AG, Reinach, Switzerland) against the most common viral pathogens of ornamentals: Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), and Impatiens necrotic spot virus (INSV) (1,2). Commercial positive and negative controls were included in each ELISA. Serological analyses showed that all plants were positive for CMV and negative for TSWV and INSV. The ELISA-positive sample (isolate 1-12) was mechanically inoculated onto five plants each of three test species as well as of healthy young P. tuisana using 0.01 M phosphate buffer (pH 7). Chlorotic local lesions on Chenopodium quinoa and severe mosaic and leaf malformations were observed on all inoculated Nicotiana tabacum ‘Samsun’ and N. glutinosa. Also, the virus was successfully mechanically transmitted to P. tuisana that reacted with symptoms identical to those observed on the original host plants. All mechanically inoculated plants were positive for CMV in DAS-ELISA. For further confirmation of CMV infection, reverse transcription (RT)-PCR was performed on extracts made from symptomatic P. tuisana, N. tabacum ‘Samsun,’ and N. glutinosa leaf materials. Total RNAs were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was carried out using One-Step RT-PCR Kit (Qiagen). A CMV-specific primer pair, CMVCPfwd and CMVCPrev (3), which amplifies an 871-bp fragment of the entire coat protein (CP) gene and part of 3′- and 5′-UTRs, were used for both amplification and sequencing. Total RNAs obtained from the Serbian CMV isolate (HM065510) and healthy P. tuisana were used as positive and negative controls, respectively. A product of the correct predicted size was obtained in all naturally and mechanically infected plants, as well as positive control. No amplicon was recorded in the healthy control. The amplified product derived from isolate 1-12 was purified (QIAquick PCR Purification Kit, Qiagen), directly sequenced in both directions, deposited in GenBank (KC505441), and analyzed by MEGA5 software (4). Sequence comparison of the complete CP gene (657 nt) revealed that the Serbian isolate 1-12 shared the highest nucleotide identity of 99.1% (99.5% amino acid identity) with the Japanese isolate (AB006813). To our knowledge, this is the first report on the occurrence of CMV in P. tuisana in Serbia. This is also an important discovery since P. tuisana is commonly grown together with other ornamental hosts of CMV, and thus could represent a serious threat for future expansion of CMV in the greenhouse floriculture industry in Serbia. References: (1) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (2) S. Flasinski et al. Plant Dis. 79:843, 1995. (3) K. Milojevic et al. Plant Dis. 96:1706, 2012. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

Download Full-text

First Report of Cucumber mosaic virus in Agastache rugosa in China

Plant Disease ◽

10.1094/pdis-04-14-0392-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1589-1589

Author(s):

J. Du ◽

C. M. Zhang ◽

Y. B. Niu

Keyword(s):

Coat Protein ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Consensus Sequence ◽

Tomato Mosaic Virus ◽

Shanxi Province ◽

Agastache Rugosa ◽

First Report ◽

Pcr Products ◽

Das Elisa

Agastache rugosa (family Lamiaceae) is one of the most common herbs of traditional Chinese medicine in China, and the species increasingly gains popularity on the international market. In June 2012, typical mosaic symptoms were observed on many A. rugosa plants in a field in Shanxi Province. The incidence of this disease reached more than 60% in a 2.6-ha field. Seven symptomatic plants were tested by DAS-ELISA using monoclonal antibodies specific for Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV), and Tomato mosaic virus (ToMV); all antisera used in DAS-ELISA were generated and validated in our laboratory. CMV was found in all seven samples tested, but not TMV and ToMV. Double-stranded RNAs (dsRNA) extracted from infected leaves were used as templates in the subsequent two-step RT-PCR reaction (1). In order to further confirm the presence of CMV, a pair of specific primers (forward: 5′-ACGTCGACCATGGACAAATC-3′, and reverse: 5′-TACCCGGGTCAGACTGGTAGCACC-3′) based on the coat protein gene sequence of CMV were used to amplify PCR products of the expected size (657 bp) from ELISA-positive samples (2). These PCR products were cloned into pUCm-T Vector (Sangon Biotech, Shanghai) and sequenced. Five independent clones have been sequenced to obtain the consensus sequence. This consensus sequence (GenBank Accession No. JQ403529) was compared with other CMV sequences available in GenBank using DNAMAN. The partial CMV coat protein sequence showed the highest 97.9% nucleotide identity with a subgroup IB CMV isolate from China (DQ459481). To our knowledge, this is the first report of the natural occurrence of CMV on A. rugosa. References: (1) M. Krajacić et al. J. Chromatogr. A. 1144:111, 2007. (2) F. Li et al. J. Zhejiang Univ. 26:261, 2000.

Download Full-text

First Report of Cucumber mosaic virus in Tulipa sp. in Serbia

Plant Disease ◽

10.1094/pdis-05-14-0527-pdn ◽

2014 ◽

Vol 98 (10) ◽

pp. 1449-1449 ◽

Cited By ~ 1

Author(s):

K. Milojević ◽

I. Stanković ◽

A. Vučurović ◽

D. Nikolić ◽

D. Ristić ◽

...

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

The United States ◽

Post Inoculation ◽

Rt Pcr ◽

First Report ◽

Cp Gene ◽

Elisa Testing ◽

Negative Controls ◽

Das Elisa

Tulips (Tulipa sp. L.), popular spring-blooming perennials in the Liliaceae family, are one of the most important ornamental bulbous plants, which have been cultivated for cut flower, potted plant, garden plant, and for landscaping. In May 2013, during a survey to determine the presence of Cucumber mosaic virus (CMV, Cucumovirus, Bromoviridae) on ornamentals in Serbia, virus-like symptoms, including the presence of bright streaks, stripe and distortion of leaves, and reduced growth and flower size, were observed in an open field tulip production in the Krnjaca locality (a district of Belgrade, Serbia). Disease incidence was estimated at 20%. Symptomatic tulip plants were collected and tested for the presence of CMV by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kit (Bioreba, AG, Reinach, Switzerland). Commercial positive and negative controls were included in each ELISA. Of the six tulip plants tested, all were positive for CMV. In bioassay, five plants of each Chenopodium quinoa, Nicotiana tabacum ‘Samsun,’ and N. glutinosa were mechanically inoculated with sap from selected ELISA-positive sample (79-13) using 0.01 M phosphate buffer (pH 7). Chlorotic local lesions on C. quinoa, and severe mosaic and leaf malformations on N. tabacum ‘Samsun’ and N. glutinosa, were observed 5 and 14 days post-inoculation, respectively. All mechanically inoculated plants were positive for CMV in DAS-ELISA testing. For further confirmation of CMV presence in tulip, total RNAs from all ELISA-positive symptomatic tulip plants were extracted with the RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Reverse transcription (RT)-PCR was performed with the One-Step RT-PCR Kit (Qiagen) using specific primer pair CMVCPfwd and CMVCPrev (1), which flank conserved fragment of the RNA3 including the entire coat protein (CP) gene and part of 3′- and 5′-UTRs. Total RNAs obtained from the Serbian watermelon CMV isolate (GenBank Accession No. JX280942) and healthy tulip leaves served as the positive and negative controls, respectively. The RT-PCR products of 871 bp were obtained from all six samples that were serologically positive to CMV, as well as from the positive control. No amplicon was recorded in the healthy control. The amplified product which derived from isolate 79-13 was purified (QIAquick PCR Purification Kit, Qiagen), directly sequenced in both directions using the same primer pair as in RT-PCR, deposited in GenBank (KJ854451), and analyzed by MEGA5 software (4). Sequence comparison of the complete CP gene (657 nt) revealed that the Serbian isolate 79-13 shared the highest nucleotide identity of 99.2% (99% amino acid identity) with CMV isolates from Japan (AB006813) and the United States (S70105). To our knowledge, this is the first report on the occurrence of CMV causing mosaic on Tulipa sp. in Serbia. Taking into account vegetative reproduction of tulips and the large scale of international trade with tulip seeding material, as well as wide host range of CMV including a variety of ornamentals (2,3), this is a very important discovery representing a serious threat for the floriculture industry in Serbia. References: (1) K. Milojević et al. Plant Dis. 96:1706, 2012. (2) M. Samuitienė and M. Navalinskienė. Zemdirbyste-Agriculture 95:135, 2008. (3) D. Sochacki. J. Hortic. Res. 21:5, 2013. (4) K. Tamura et al. Mol. Biol. Evol. 28:2731, 2011.

Download Full-text