scholarly journals First Report of Pepino mosaic virus on Natural Hosts

Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1292-1292 ◽  
Author(s):  
C. Jordá ◽  
A. Lázaro Pérez ◽  
P. V. Martínez Culebras ◽  
A. Lacasa
Keyword(s):  
Mosaic Virus ◽  
Solanum Nigrum ◽  
Enzyme Linked Immunosorbent Assay ◽  
New Disease ◽  
Nicotiana Glauca ◽  
Pepino Mosaic Virus ◽  
Tomato Plants ◽  
First Report ◽  
Sonchus Oleraceus ◽  
Chenopodium Murale

Pepino mosaic virus (PepMV) is a potexvirus recently identified as the causal agent of a new disease occurring in protected tomato (Lycopersicon esculentum Mill.) crops in the Netherlands (2). PepMV has been subsequently identified in England, Germany, Italy, Morocco, Portugal, and Spain. The new disease has become a serious problem for tomato production in Europe. Most infected tomato plants expressed leaf distortion, chlorosis, and yellow mosaic. Other plants expressed mosaic and bubbling of the leaf surface. Tomato fruits showing severe discoloration and mosaic were observed in protected tomato crops. Symptoms attenuated in tomato plants as the ambient temperature increased. At present, only Solanum muricatum Ait. (Peruvian pepino) and L. esculentum are affected by PepMV.To determine possible reservoir hosts for this virus, 70 samples from Amaranthus sp., A. viridis (L.) Britton et al., Chenopodium murale L., Convolvulus arvensis L., Malva parviflora L., Nicotiana glauca Grah., Polypogon monspeliensis (L.) Desf., Senecio vulgaris L., Sisybrium sp., Solanum nigrum L., and Sonchus oleraceus L. were analyzed. The plants were collected around greenhouses affected by PepMV from different regions in Spain (Murcia and Canary Islands). The samples were analyzed for PepMV by double-antibody sandwich enzyme-linked immunosorbent assay with a commercial antiserum (DSMZ AS-0554, Biologische Bundesantstal, Braunschweig, Germany). Only Amaranthus sp., M. parviflora, N. glauca, Solanum nigrum, and Sonchus oleraceus tested postive. The presence of PepMV in these weed species was confirmed by electron microscopy and reverse transcription-polymerase chain reaction using degenerate primers for potexvirus (1). All the hosts analyzed were asymptomatic. However, symptoms were reproduced by mechanically inoculating tomato plants with sap from naturally infected weeds. To our knowledge, this is the first report of natural infection of weeds by PepMV. References: (1) A. Gibbs et al. J. Virol. Methods 74:67, 1998. (2) R. A. A. Van der Vlugt et al. Plant Dis. 84:103, 2000.

scholarly journals First Report of Pepino mosaic virus on Tomato in Spain

Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1292-1292 ◽  
Author(s):  
C. Jordá ◽  
A. Lázaro Pérez ◽  
P. Martínez-Culebras ◽  
P. Abad ◽  
A. Lacasa ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Canary Islands ◽  
Plant Protection ◽  
Polyclonal Antibodies ◽  
Datura Stramonium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chenopodium Amaranticolor ◽  
Pepino Mosaic Virus ◽  
Tomato Plants ◽  
First Report

At the beginning of 2000, a damaging disease developed on protected tomato (Lycopersicon esculentum) crops grown in polyethylene greenhouses in different regions of Spain. Production losses were estimated at 15 to 80%. The tomato plants showed a variety of symptoms. The most common symptoms were leaf distortion, chlorosis, and mosaic. Some plants showed a dark green mosaic and bubbling of the leaf surface. Green striations were also observed on the stem and sepals. Most of the diseased plants had discolored fruits. Symptoms decreased as environmental temperature increased. The involvement of Pepino mosaic virus (PepMV) was suspected. To identify the etiological agent, ≈500 symptomatic tomato plants were collected from several locations in Alicante, Murcia, Almeria and the Canary Islands. Flexuous viral particles 510 nm long were observed by transmission electron microscopy, suggesting the presence of a potexvirus in the tissue extracts analyzed. All samples were tested by ELISA (enzyme-linked immunosorbent assay), using polyclonal antibodies to Narcissus mosaic virus (Adgen, Auchincriuve, Scotland), a virus serologically related to PepMV, and two antisera specific to PepMV (Adgen, Scotland and DMSZ, Braunschweig, Germany). PepMV was detected in 35% of the samples. Like PepMV, the virus infected (as confirmed by ELISA) greenhouse-grown Datura stramonium, Nicandra physalodes, Nicotiana benthamiana, N. clevelandii, Solanum tuberosum, and Vigna sinensis and did not infect Capsicum anuum, Cucumis sativus, Chenopodium amaranticolor, C. quinoa, Petunia × hybrida, Phaseolus vulgaris, Physalis floridana, N. glutinosa, N. rustica, or N. tabacum. The virus did infect Gomphrena globosa, which normally is not infected by PepMV. The first report of PepMV was on pepino (Solanum muricatum) in Peru in 1974 (1), but this virus has been recently reported in the Netherlands, England, Germany, and France on protected tomato crops (2). To our knowledge, this is the first report of PepMV in Spain, including the Canary Islands. References: (1) R. A. C. Jones et al. Ann. Appl. Biol. 94:61, 1980. (2) European and Mediterranean Plant Protection Organisation (EPPO). Alert List Viruses. On-line publication/2000/003.

scholarly journals New Natural Hosts of Pepino mosaic virus in Spain

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 906-906 ◽  
Author(s):  
M. C. Córdoba ◽  
Ll. Martínez-Priego ◽  
C. Jordá
Keyword(s):  
Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Weed Species ◽  
Convolvulus Arvensis ◽  
Pepino Mosaic Virus ◽  
Chenopodium Murale ◽  
Natural Hosts ◽  
Weed Hosts ◽  
Heliotropium Europaeum ◽  
Diplotaxis Erucoides

Pepino mosaic virus (PepMV) was first detected in Spain in 2000 (1). The virus infects tomato (Lycopersicon esculentum Mill.) crops and causes a variety of symptoms including leaf distortion, chlorosis, mosaic, blistering of the leaf surface, green striations on the stem and sepals, and fruit discoloration. PepMV is present along the southern and eastern regions of Spain (provinces of Granada, Almeria, Murcia, Alicante, Valencia, and Barcelona), Balearic, and the Canary Islands. In the summer and autumn of 2001 and 2002, virus-like symptoms were observed in native plants growing in or around tomato fields in Murcia and Almeria provinces. To study the alternate hosts that may serve as virus reservoirs, 62 samples of 42 common weed species, including asymptomatic plants, were collected and analyzed for PepMV using double-antibody sandwich enzyme-linked immunosorbent assay with a commercial antiserum (DSMZ As-0554; Biologische Bundesantstal, Braunschweig, Germany). The following weed hosts tested positive for PepMV: Bassia scoparia (L.) Voss., Calystegia sepium (L.) R.Br., Chenopodium murale L., Convolvulus althaeoides L., Convolvulus arvensis L., Conyza albida Willd. ex Spreng., Coronopus sp., Diplotaxis erucoides (L.) DC, Echium creticum L., E. humile Desf., Heliotropium europaeum L., Moricandia arvensis (L.) DC., Onopordum sp., Piptatherum multiflorum (Cav.) Beauv., Plantago afra L., Rumex sp., Sisymbrium irio L., Sonchus tenerrimus L., and Taraxacum vulgare (Lam.) Schrank. The presence of PepMV in these weed species was confirmed using reverse transcription-polymerase chain reaction with primers specific for PepMV (2). Although the number of samples examined may be insufficient to assess precisely the role of weed reservoirs in outbreaks of PepMV, these findings reveal potential virus sources and contribute to further understanding of PepMV epidemiology in Spain. References: (1) C. Jordá et al. Plant Dis. 85:1292, 2001. (2) P. Martínez-Culebras et al. Eur. J. Plant Pathol. 108:887, 2002.

scholarly journals First Report of Pepino mosaic virus Infecting Greenhouse Cherry Tomatoes in Greece

Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 78-78 ◽  
Author(s):  
K. E. Efthimiou ◽  
A. P. Gatsios ◽  
K. C. Aretakis ◽  
L. C. Papayiannis ◽  
N. I. Katis
Keyword(s):  
Mosaic Virus ◽  
Leaf Tissue ◽  
Tomato Mosaic Virus ◽  
Sequencing Analysis ◽  
Rt Pcr ◽  
Pepino Mosaic Virus ◽  
Tomato Plants ◽  
First Report ◽  
Negative Controls ◽  
Das Elisa

Pepino mosaic virus (PepMV) (genus Potexvirus, family Flexiviridae) is a mechanically transmitted virus that has emerged as a significant problem of greenhouse tomato crops in Europe and around the world during the past 10 years (1). In spring of 2010, mosaic symptoms were observed on leaves of cherry tomato (Lycopersicon esculentum var. cerasiforme) greenhouse crops (hybrids Shiren, Tomito, and Rubino top) in the areas of Drymos and Vonitsa, located at Aitoloakarnania Prefecture, in Greece. A total of 63 tomato samples (55 from symptomatic and 8 from asymptomatic plants) were collected from 11 greenhouses where disease incidence ranged from 10 to 20%. All samples were tested by double-antibody sandwich (DAS)-ELISA using polyclonal antibodies from BIOREBA, AG (Reinach, Switzerland) for the presence of PepMV, Cucumber mosaic virus (CMV), and Tomato mosaic virus (ToMV). Leaf tissue from PepMV-, CMV-, and ToMV-infected samples and virus-free tomato plants were included in all tests as positive and negative controls, respectively. Results showed that 53 symptomatic samples collected from all greenhouses were infected with PepMV and two were co-infected with PepMV and CMV. Total RNA was extracted from all infected plants with a commercially available kit (Qiagen, Hilden, Germany) and amplified by conventional and real-time reverse transcription (RT)-PCR, using previously reported protocols (2). Positive and negative controls were also included in each assay. The 200-bp amplified PCR fragments of Triple Gene Block 3 (TGB3) obtained from five infected samples were purified and both strands were sequenced. Sequencing data were analyzed, deposited in the GenBank, and compared with other reported sequences. In addition, leaf tissue from five samples infected with only PepMV was used for mechanical inoculation of four plants of Nicotiana glutinosa, N. benthamiana, and tomato (L. esculentum FA 179 hybrid) plants. As negative controls, two plants from each species were used. Sequencing analysis showed that all five PepMV sequences were identical (GenBank Accession Nos. FR686904 to FR686908) and possessed 100% identity PepMVstrain CH2 (DQ000985). Inoculation results showed that the virus was successfully transmitted to N. benthamiana and tomato plants which developed mosaic symptoms, and tested positive by DAS-ELISA and RT-PCR. N. glutinosa plants did not develop any symptoms and were found to be free of PepMV when tested by DAS-ELISA and RT-PCR. To our knowledge, this is the first report of PepMV in Greece. Further studies on the disease prevalence and incidence and its economic impact on tomato production are required. PepMV is currently under quarantine status in the EU and therefore new protective measures should be recommended to prevent the spread of PepMV to other regions of Greece. References: (1) I. M. Hanssen and B. P. H. J. Thomma. Mol. Plant Pathol. 11:179, 2010. (2) K. S. Ling et al. J. Virol. Methods 144:65, 2007.

scholarly journals First Report of Cucumber mosaic virus Subgroup II Infecting Lycopersicon esculentum in India

Plant Disease ◽  
2006 ◽  
Vol 90 (11) ◽  
pp. 1457-1457 ◽  
Author(s):  
N. Sudhakar ◽  
D. Nagendra-Prasad ◽  
N. Mohan ◽  
K. Murugesan
Keyword(s):  
Coat Protein ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Tamil Nadu ◽  
Indian Subcontinent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Leaf ◽  
Polymorphism Analysis ◽  
First Report ◽  
Subgroup Ii

During a survey in January 2006 near Salem in Tamil Nadu (south India), Cucumber mosaic virus was observed infecting tomatoes with an incidence of more than 70%. Plants exhibiting severe mosaic, leaf puckering, and stunted growth were collected, and the virus was identified using diagnostic hosts, evaluation of physical properties of the virus, compound enzyme-linked immunosorbent assay (ELISA) (ELISA Lab, Washington State University, Prosser), reverse-transcription polymerase chain reaction (RT-PCR), and restriction fragment length polymorphism analysis (DSMZ, S. Winter, Germany). To determine the specific CMV subgroup, total RNA was extracted from 50 infected leaf samples using the RNeasy plant RNA isolation kit (Qiagen, Hilden, Germany) and tested for the presence of the complete CMV coat protein gene using specific primers as described by Rizos et al. (1). A fragment of the coat protein was amplified and subsequently digested with MspI to reveal a pattern of two fragments (336 and 538 bp), indicating CMV subgroup II. No evidence of mixed infection with CMV subgroup I was obtained when CMV isolates representing subgroups I (PV-0419) and II (PV-0420), available at the DSMZ Plant Virus Collection, were used as controls. Only CMV subgroup I has been found to predominantly infect tomato in the Indian subcontinent, although Verma et al. (2) identified CMV subgroup II infecting Pelargonium spp., an ornamental plant. To our knowledge, this is the first report of CMV subgroup II infecting tomato crops in India. References: (1) H. Rizos et al. J. Gen. Virol. 73:2099, 1992. (2) N. Verma et al. J. Biol. Sci. 31:47, 2006.

scholarly journals Short communication: Pepino mosaic virus, a new threat for Serbia’s tomatoes

2020 ◽  
Vol 18 (4) ◽  
pp. e10SC05
Author(s):  
Ivana Stankovic ◽  
Ana Vucurovic ◽  
Katarina Zecevic ◽  
Branka Petrovic ◽  
Danijela Ristic ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Rt Pcr ◽  
Tomato Production ◽  
Pepino Mosaic Virus ◽  
Tomato Plants ◽  
Rapid Spread ◽  
Infected Tomato ◽  
Dark Green ◽  
Relationship Of ◽  
Das Elisa

Aim of study: To report the occurrence of Pepino mosaic virus (PepMV) on tomato in Serbia and to genetically characterize Serbian PepMV isolates.Area of study: Tomato samples showing virus-like symptoms were collected in the Bogojevce locality (Jablanica District, Serbia).Material and methods: Collected tomato samples were assayed by DAS-ELISA using antisera against eight economically important or quarantine tomato viruses. Three selected isolates of naturally infected tomato plants were mechanically transmitted to tomato ‘Novosadski jabučar’ seedlings. For confirmation of PepMV infection, RT-PCR was performed using specific primers PepMV TGB F/PepMV UTR R. Maximum-likelihood phylogenetic tree was constructed with 47 complete CP gene sequences of PepMV to determine the genetic relationship of Serbian PepMV isolates with those from other parts of the world.Main results: The results of DAS-ELISA indicated the presence of PepMV in all tested samples. Mechanically inoculated ‘Novosadski jabučar’ seedlings expressed yellow spots and light and dark green patches, bubbling, and curled leaves. All tested tomato plants were RT-PCR positive for the presence of PepMV. The CP sequence analysis revealed that the Serbian PepMV isolates were completely identical among themselves and shared the highest nucleotide identity of 95.1% (99.2% aa identity) with isolate from Spain (FJ263341). Phylogenetic analysis showed clustering of the Serbian PepMV isolates into CH2 strain, but they formed separate subgroup within CH2 strain.Research highlights: This is the first data of the presence of PepMV in protected tomato production in Serbia. Considering increased incidence and rapid spread in Europe, the presence of PepMV on tomato could therefore represent serious threat to this valuable crop in Serbia.

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).

High population densities of Macrolophus pygmaeus on tomato plants can cause economic fruit damage: interaction with Pepino mosaic virus ?

2015 ◽  
Vol 72 (7) ◽  
pp. 1350-1358 ◽  
Author(s):  
Rob Moerkens ◽  
Els Berckmoes ◽  
Veerle Van Damme ◽  
Nelia Ortega-Parra ◽  
Inge Hanssen ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
High Population ◽  
Population Densities ◽  
Pepino Mosaic Virus ◽  
Tomato Plants ◽  
Macrolophus Pygmaeus ◽  
Fruit Damage

FIRST REPORT OF PEPINO MOSAIC VIRUS (PEPMV) IN CROATIA

2012 ◽  
pp. 321-325
Author(s):  
A. Novak ◽  
J. Milanović ◽  
V. Kajić
Keyword(s):  
Mosaic Virus ◽  
Pepino Mosaic Virus ◽  
First Report

scholarly journals Pepino mosaic virus on Tomato Seed: Virus Location and Mechanical Transmission

Plant Disease ◽  
2008 ◽  
Vol 92 (12) ◽  
pp. 1701-1705 ◽  
Author(s):  
Kai-Shu Ling
Keyword(s):  
Mosaic Virus ◽  
Large Scale ◽  
Seed Transmission ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mechanical Transmission ◽  
Tomato Seed ◽  
Tomato Production ◽  
Pepino Mosaic Virus ◽  
Infectivity Assay ◽  
Seed Coats

In just a few years, Pepino mosaic virus (PepMV) has become a major threat to greenhouse tomato production around the world. Although tomato seed is suspected to spread the disease, its importance as an initial virus inoculum for PepMV has not been established. To determine the potential for seed transmission, a tomato seed lot highly contaminated with PepMV was used for large-scale seedling grow-out tests. None of 10,000 grow-out seedlings was infected as determined by symptom expression, enzyme-linked immunosorbent assay (ELISA), or infectivity assay on Nicotiana benthamiana. Even though PepMV was not seed transmitted on tomato, the virus was effectively transmitted to tomato and N. benthamiana seedlings through mechanical transmission with seed extract. To examine the exact location where PepMV particles accumulated on the tomato seed, seed coats and embryos were carefully isolated and tested separately by ELISA, real-time RT-PCR, and bioassay on N. benthamiana. PepMV was detected in the seed coat fraction in both immature and mature tomato seeds, but not in the embryo. However, in N. benthamiana, the virus was neither seedborne nor seed-transmitted. Because PepMV is seedborne in tomato, efficient mechanical transmission of PepMV from the virus-contaminated tomato seed to seedlings could initiate a disease epidemic in a new tomato growing area. Thus, it is important to plant certified tomato seed that has been tested free of PepMV.

scholarly journals First Report of Chilli veinal mottle virus Infecting Tomato (Solanum lycopersicum) in China

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1589-1589 ◽  
Author(s):  
F.-F. Zhao ◽  
D.-H. Xi ◽  
J. Liu ◽  
X.-G. Deng ◽  
H.-H. Lin
Keyword(s):  
Mosaic Virus ◽  
Southwest China ◽  
Sichuan Province ◽  
Mottle Virus ◽  
Rt Pcr ◽  
Tomato Plants ◽  
First Report ◽  
Two Samples ◽  
Leaf Distortion ◽  
Chilli Veinal Mottle Virus

Chilli veinal mottle virus (ChiVMV), a potyvirus, is widespread over the world. In China, it was first reported in chili pepper (Capsicum annuum) in Hainan Province (south China) in 2006 (2). Subsequently, it was reported in tobacco (Nicotiana tabacum) in Yunnan Province (southwest China) in 2011 (1). Sichuan Province is one of the largest vegetable producing areas of China. In May 2012, tomatoes with leaves displaying virus-infected symptoms like mottling, mosaic, narrowing, or curling were observed in several fields of Chengdu, eastern Sichuan Province, southwest China. Of the 20 fields we investigated, four fields with 90% tomato plants were infected. During 2012 and 2013, six samples were collected from symptomatic tomato leaves based on different symptoms and locations. All six samples were assayed by western blotting using polyclonal antisera (Cucumber mosaic virus [CMV], Tobacco mosaic virus [TMV]) obtained from Agdia (Elkhart) and one antiserum to ChiVMV obtained from Yunnan Academy of Agricultural Science (China). Two samples from Pengzhou and one sample from Shuangliu exhibiting mosaic leaves were positive for TMV, one sample from Pixian exhibiting narrowing leaves was positive for CMV, and the other two samples from Shuangliu exhibiting mottle and leaf distortion were positive for ChiVMV. Total RNAs was extracted from all six samples and healthy tomato leaves using Trizol reagent (Invitrogen), First-strand cDNA synthesis primed with oligo(dT) by SuperScript III Reverse Transcriptase (Invitrogen). RT-PCR was performed using primer pairs ChiVMV-CP F (5′-GCAGGAGAGAGTGTTGATGCTG-3′) and ChiVMV–CP R (5′-(T)16AACGCCAACTATTG-3′), which were designed to direct the amplification of the entire capsid protein (CP) gene and 3′ untranslated region (3′-UTR) of ChiVMV (GenBank Accession No. KC711055). The expected 1,166-bp DNA fragment was amplified from the two tomato samples from Shuangliu that were positive for ChiVMV in the western blot tests, but not from the others. The obtained fragments were purified and cloned into the PMD18-T vector (TaKaRa) and sequenced. The sequencing results showed that the two ChiVMV isolates from tomato in Shuangliu were identical (KF738253). Nucleotide BLAST analysis revealed that this ChiVMV isolate shared ~84 to 99% nucleotide identities with other ChiVMV isolates available in GenBank (KC711055 to KF220408). To fulfill Koch's postulates, we isolated this virus by three cycle single lesion isolation in N. tabacum, and mechanically inoculated it onto tomato leaves. The same mottle and leaf distortion symptoms in systemic leaves were observed. Subsequent RT-PCR, fragment clone, and sequence determination tests were repeated and the results were the same. All the evidence from these tests revealed that the two tomato plants were infected by ChiVMV. To our knowledge, this is the first report of ChiVMV naturally infecting tomato in China. It shows that ChiVMV is spreading in China and is naturally infecting a new solanaceous crop in the southwest area, and the spread of the virus may affect tomato crop yields in China. Thus, it is very important to seek an effective way to control this virus. References: (1) M. Ding et al. Plant Dis. 95:357, 2011. (2) J. Wang et al. Plant Dis. 90:377, 2006.

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