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

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.

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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.

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First Report of Zucchini yellow mosaic virus, Watermelon mosaic virus, and Cucumber mosaic virus in Bottlegourd (Lagenaria siceraria) in Serbia

Plant Disease ◽

10.1094/pd-90-0380a ◽

2006 ◽

Vol 90 (3) ◽

pp. 380-380 ◽

Cited By ~ 8

Author(s):

N. Dukić ◽

B. Krstić ◽

I. Vico ◽

J. Berenji ◽

B. Duduk

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Zucchini Yellow Mosaic Virus ◽

Yellow Mosaic Virus ◽

Watermelon Mosaic Virus ◽

Mechanical Transmission ◽

Lagenaria Siceraria ◽

Indicator Plants ◽

First Report ◽

Das Elisa

During a cucurbit disease survey in August 2004, severe symptoms resembling those caused by viruses were observed on bottlegourd (Lagenaria siceraria (Molina) Standl.) in the Vojvodina region of Serbia. Symptoms included stunting, mosaic, green veinbanding, blistering, yellowing, chlorotic spots, leaf deformation, and fruit distortion. Leaf samples from 25 symptomatic plants were collected from two localities for virus identification using mechanical transmission and serological testing. Crude sap extract from leaf samples was mechanically inoculated onto bottlegourd and pumpkin (Cucurbita pepo) under greenhouse conditions. Field-collected bottlegourd and inoculated plants were tested using double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA). Positive reactions were obtained on collected and inoculated plants with polyclonal antiserum (Loewe Biochemica, Sauerlach, Germany) to Zucchini yellow mosaic virus(ZYMV) in 23 samples, with antiserum to Watermelon mosaic virus (WMV) in eight samples, and with antiserum to Cucumber mosaic virus (CMV) in seven samples. Each of the three viruses was detected in single as well as in mixed infections with the other two viruses. Biological characterization of viruses detected in single infections was done on the following indicator plants: Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Cucumis melo, Citrullus lanatus, Nicotiana glutinosa, and N. tabacum cv. Samsun. The symptoms observed on indicator plants for each isolate corresponded to the results of DAS-ELISA (2,3). All three viruses are known to be important pathogens of cucurbit plants and were previously reported in pumpkin in Serbia (1). To our knowledge, this is the first report of ZYMV, WMV, and CMV in bottlegourd in Serbia. References: (1) N. Dukić et al. J. Agric. Sci. 47:149, 2002. (2) D. E. Lesemann et al. Phytopathol. Z. 108:304, 1983. (3) H. Rahimian and K. Izadpanah. Phytopathol. Z. 92:305, 1978.

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First Report of Cucumber mosaic virus in Desert Rose in Taiwan

Plant Disease ◽

10.1094/pdis-11-11-0917 ◽

2012 ◽

Vol 96 (4) ◽

pp. 593-593 ◽

Cited By ~ 1

Author(s):

Y. K. Chen ◽

Y. S. Chang ◽

Y. W. Lin ◽

M. Y. Wu

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Chenopodium Quinoa ◽

Rabbit Antiserum ◽

Serological Tests ◽

Reading Frame ◽

First Report ◽

Reverse Transcription Pcr ◽

Wilt Virus ◽

Line Patterns

Desert rose (Adenium obesum (Forssk.) Roem. & Schult, family Apocynaceae) is native to southeastern Africa, and is a perennial potted ornamental with colorful flowers that are popular in Taiwan. Symptoms of mosaic and chlorotic ringspots and line patterns on leaves were observed in July 2010, on all eight plants in a private garden in Potzu, Chiayi, Taiwan. Spherical virus particles with a diameter of approximately 28 nm were observed in crude sap prepared from symptomatic leaves. Virus culture was established by successive local lesion isolation in Chenopodium quinoa and was maintained in the systemic host Nicotiana tabacum van Hicks. The virus was mechanically transmissible to indicator plants and induced symptoms similar to those incited by Cucumber mosaic virus (CMV). Observed symptoms included local lesions on inoculated leaves of C. amaranticolor and systemic mosaic in Cucumis sativus, Lycopersicon esculentum, N. benthamiana, N. glutinosa, and N. rustica. On N. tabacum, necrotic ringspots developed on inoculated leaves followed by systemic mosaic. Serological tests using ELISA assays and western blotting indicated that the virus reacted positively to a rabbit antiserum prepared to CMV (4). Amplicons of an expected size (1.1 kb) were obtained in reverse transcription-PCR with primers specific to the 3′-half of CMV RNA 3 (3) using total RNA extracted from infected desert rose and N. tabacum. The amplified cDNA fragment was cloned and sequenced (GenBank Accession No. AB667971). Nucleotide sequences of the coat protein open reading frame (CP ORF) (657 nt) had 92 to 96% and 76 to 77% sequence identity to those of CMV in subgroups I (GenBank Accession Nos. NC_001440, D00385, M57602, D28780, and AB008777) and II (GenBank Accession Nos. L15336, AF127976, AF198103, and M21464), respectively. Desert roses infected by Tomato spotted wilt virus (TSWV) (1) and CMV (2) have been reported previously. In spite of the plants showing mosaic symptoms similar to that caused by CMV (2) and chlorotic ringspots and line patterns caused by TSWV (1), only CMV was detected in and isolated from these infected desert roses. However, the possibility of mixed infection of CMV and other viruses were not excluded in this research. To our knowledge, this is the first report of CMV infection in desert rose plants occurring in Taiwan. References: (1) S. Adkins and C. A. Baker. Plant Dis. 89:526, 2005. (2) C. A. Baker et al. Plant Dis. 87:1007, 2003. (3) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (4) Y. K. Chen and C. C. Yang. Plant Dis. 89:529, 2005.

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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.

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First Report of Cucumber mosaic virus in Paeonia lactifera in France

Plant Disease ◽

10.1094/pdis-94-6-0790c ◽

2010 ◽

Vol 94 (6) ◽

pp. 790-790 ◽

Cited By ~ 3

Author(s):

L. Cardin ◽

J. P. Onesto ◽

B. Moury

Keyword(s):

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Polyclonal Antibodies ◽

Tobacco Rattle Virus ◽

Meristem Culture ◽

Cut Flowers ◽

Leaf Extracts ◽

First Report ◽

The Family ◽

Das Elisa

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.

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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.

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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.

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First Report of Cucumber mosaic virus Infecting Blephilia hirsuta in North America

Plant Disease ◽

10.1094/pdis-94-8-1070c ◽

2010 ◽

Vol 94 (8) ◽

pp. 1070-1070 ◽

Cited By ~ 1

Author(s):

B. Poudel ◽

A. G. Laney ◽

I. E. Tzanetakis

Keyword(s):

North America ◽

Endangered Species ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Chenopodium Quinoa ◽

Dot Blot ◽

Degenerate Primers ◽

First Report ◽

Wide Range ◽

Symptomatic Tissue

Blephilia hirsuta (Pursh) Benth. var. hirsuta, an ornamental plant known as hairy pagoda or hairy wood mint (Lamiaceae), is native to eastern North America and is listed as an endangered species or a species of special concern in several northeastern states ( http://www.ct.gov/dep/cwp/view.asp?a=2702&q=323482&depNav_GID=1628 and http://www.mass.gov/dfwele/dfw/nhesp/species_info/mesa_list/mesa_list.htm ). B. hirsuta, grown as an ornamental on the University of Arkansas campus in Fayetteville, exhibited mottling symptoms indicative of viral infection. Double-stranded RNA extractions (3) yielded four bands of approximately 3.2, 2.9, 2.2, and 0.9 kb, a pattern identical to that of Cucumber mosaic virus (CMV [2]). Nicotiana benthamiana and Chenopodium quinoa seedlings were mechanically inoculated with sap from symptomatic tissue. N. benthamiana inoculated plants were stunted and developed systemic mosaic and C. quinoa inoculated plants developed local lesions, whereas mock inoculated plants remained symptomless. Dot-blot and indirect ELISA using antisera against CMV (developed by H. A Scott) gave strong reactions when testing symptomatic tissue from B. hirusta, N. benthamiana, and C. quinoa compared with no reaction for symptomless plants. Total nucleic acid extractions (4) from symptomatic tissue was subjected to reverse transcription-PCR using Cucumovirus degenerate primers (1). An amplicon of approximately 940 bases was obtained and sequenced. The sequence, deposited in GenBank under Accession No. GU453918, confirmed the results of the immunological assays that B. hirsuta was infected with CMV. The nucleotide identities between the B. hirsuta isolate and those of the Fny CMV group exceeded 98%. To our knowledge, this is the first report of CMV infecting B. hirsuta, not only in North America, but globally. This finding has major implications for the ornamental industry and the viability of the endangered species. Given the wide range of CMV, B. hirsuta may act as a reservoir for the virus and facilitate transmission to ornamentals and other plants. In addition, the virus may reduce host fitness and undermine the efforts to preserve the species in areas that is threatened. References: (1) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (2) I. E. Tzanetakis. Plant Dis. 93:431, 2009. (3) I. E. Tzanetakis and R. R. Martin. J. Virol. Methods 149:167, 2008. (4) I. E. Tzanetakis et al. Virus Res. 127:26, 2007.

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First Report of Cucumber mosaic virus and Turnip vein-clearing virus in Dichondra repens in France, Italy, and China

Plant Disease ◽

10.1094/pdis-93-2-0201b ◽

2009 ◽

Vol 93 (2) ◽

pp. 201-201 ◽

Cited By ~ 2

Author(s):

L. Cardin ◽

B. Delecolle ◽

B. Moury

Keyword(s):

Electron Microscope ◽

Cucumber Mosaic Virus ◽

Mosaic Virus ◽

Plantago Lanceolata ◽

Aphis Gossypii ◽

Chenopodium Quinoa ◽

Chenopodium Amaranticolor ◽

First Report ◽

Vein Clearing ◽

Local Lesions

During surveys of Dichondra repens (kidneyweed, family Convolvulaceae) turfs in public gardens of the Franco-Italian Riviera from 1993 to 2003, leaf mosaic and yellow ringspot symptoms have been observed in Antibes, Menton, Nice, and Vallauris (France) and San Remo and La Mortola (Italy). Isolates from these six locations and from two locations in China (Shanghai and Kunming) have revealed the presence of Cucumber mosaic virus (CMV) based on the behavior of a range of manually inoculated plants (1), the observation of 30 nm isometric particles in semipurified extracts of inoculated Nicotiana tabacum ‘Xanthi’ plants with the electron microscope, and positive reactions in double antibody sandwich (DAS)-ELISAs with specific polyclonal antibodies. All isolates were shown to belong to group II of CMV isolates (3) by double-immunodiffusion analysis. CMV was previously identified in D. repens in California in 1972 (4). Following isolation from local lesions on Vigna unguiculata and multiplication in ‘Xanthi’ tobacco plants, two of the isolates were used to inoculate seedlings of D. repens manually or by Aphis gossypii aphids. Two months later, all inoculated plants showed symptoms similar to those previously observed and were positive in DAS-ELISA. In 2000, a D. repens sample collected in Antibes showing similar symptoms as above, induced necrotic local lesions in inoculated ‘Xanthi’ plants in 48 h, followed by systemic mosaic symptoms typical of CMV, therefore revealing the presence of a second virus. That virus was separated from CMV in apical, noninoculated leaves of Chenopodium quinoa and then used to inoculate a range of test plants. It was infectious in most plants of the families Solanaceae (including Cyphomandra betacea) and Brassicaceae, together with in Chenopodium amaranticolor, C. quinoa, Claytonia perfoliata, Convolvulus spp. ‘Belle de jour’, Digitalis purpurea, Gomphrena globosa, Ocimum basilicum, Plantago lanceolata, and Valerianella olitoria. It induced asymptomatic systemic infections in D. repens. Numerous, rod-shaped, 300 nm long particles were observed in sap extracts of infected plants with the electron microscope, suggesting the presence of a tobamovirus. A set of primers polyvalent for tobamoviruses (2) allowed the amplification of a DNA product of approximately 800 bp through reverse transcription-PCR performed with total RNA extracts from inoculated ‘Xanthi’ plants. The DNA product was cloned and sequenced (GenBank Accession No. EU927306) revealing that the virus belonged to a tobamovirus lineage including Ribgrass mosaic virus and viruses infecting cruciferous plants (Turnip vein-clearing virus [TVCV] and Youcai mosaic virus) and was closest to TVCV (95% amino acid identity; GenBank Accession No. NC_001873). To our knowledge, this is the first report of TVCV in D. repens. References: (1) L. Cardin et al. Plant Dis. 87:200, 2003. (2) A. Gibbs et al. J. Virol. Methods 74:67, 1998. (3) M. J. Roossinck. J. Virol. 76:3382, 2002. (4) L. G. Weathers and D. J. Gumpf. Plant Dis. Rep. 56:27, 1972.

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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.

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