scholarly journals Characteristics of a potyvirus associated with a mosaic-like disease of yellow oat-grass

2010 ◽  
Vol 40 (No. 2) ◽  
pp. 37-41 ◽  
Author(s):  
L. Širlová ◽  
J. Vacke ◽  
M. Jokeš
Keyword(s):  
Host Range ◽  
Mosaic Virus ◽  
Host Plants ◽  
Indirect Elisa ◽  
Mottle Virus ◽  
Mechanical Transmission ◽  
Rt Pcr ◽  
Virus Particles ◽  
Bromus Mollis ◽  
Das Elisa

The properties of a new filamentous virus found on yellow oat-grass (suggested name <I>Trisetum flavescens mosaic virus</I> – TFMV) were compared with those of the two potyviruses <I>Oat mosaic virus (</I>OMV) and <I>Oat necrotic mottle virus</I> (ONMV). The latter viruses were chosen because their host range, virus particles and some other characteristics are similar to TFMV. Mechanical transmission of TFMV to some OMV and ONMV host plants, drop precipitation, indirect-ELISA, DAS-ELISA, SSEM and RT-PCR were used in the study. However, there was no proof that TFMV is identical with OMV or ONMV. <I>Avena abyssinica</I> and <I>Bromus mollis</I> were found to be new experimental hosts of TFMV.&nbsp;

scholarly journals Bidens mottle virus Identified in Tropical Soda Apple in Florida

Plant Disease ◽  
2007 ◽  
Vol 91 (7) ◽  
pp. 905-905 ◽  
Author(s):  
C. A. Baker ◽  
I. Kamenova ◽  
R. Raid ◽  
S. Adkins
Keyword(s):  
Host Range ◽  
Mosaic Virus ◽  
Plant Viruses ◽  
Mottle Virus ◽  
Tomato Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Product ◽  
Potyvirus Species ◽  
Tropical Soda Apple

Tropical soda apple (TSA) (Solanum viarum Dunal), a plant native to South America, was first identified in Florida in 1988 (4). It rapidly became a noxious weed in pastures throughout the state and it is known to be a reservoir for Cucumber mosaic virus, Potato leafroll virus, Potato virus Y (PVY), Tobacco etch virus (TEV), Tomato mosaic virus, and Tomato mottle virus, viruses that infect important vegetable crops in Florida (3). During a routine survey of Florida weeds during May of 2004, a TSA plant with chlorotic, young leaves found near Okeechobee, FL was determined to be infected with a potyvirus by using a commercially available enzyme linked immunosorbent assay kit (Agdia, Elkhart, IN). The results of a host range study indicated this potyvirus was neither PVY nor TEV. The virus caused local lesions in Chenopodium amaranticolor and systemic symptoms in C quinoa, Coreopsis sp. (C. A. Baker, unpublished), Helianthus annus, Nicotiana benthamiana, Petunia × hybrida, Verbena hybrida, and Zinnia elegans. It did not infect Gomphrena globosa, N. glutinosa, Pisum sativum, or Phaseolus vulgaris (1). Cylindrical inclusions consistent with those observed in plants infected with Bidens mottle virus (BiMoV) were observed in Z. elegans. Immunodiffusion tests with antiserum to BiMoV (Department of Plant Pathology, University of Florida) gave a reaction of identity with leaf extracts of the symptomatic zinnia, a known sample of BiMoV originally isolated from Bidens pilosa and a recent isolate of BiMoV from lettuce in Belle Glade, FL (C. A. Baker and R. Raid, unpublished). A partial polyprotein gene fragment (GenBank Accession No. EF467235) was amplified from total RNA of an inoculated C. quinoa plant by reverse transcription (RT)-PCR with previously described degenerate potyvirus primers (2). Analysis of the RT-PCR product sequence confirmed the host range results and indicated that the potyvirus infecting TSA was neither PVY nor TEV. However, the nucleotide and deduced amino acid sequences of a 247-bp portion of the RT-PCR product were 94 and 98% identical, respectively, with the coat protein sequence (GenBank Accession No. AF538686) of Sunflower chlorotic spot virus (SCSV). SCSV is a tentative potyvirus species described from Taiwan that is not yet recognized as an accepted species by the International Committee on Taxonomy of Viruses. On the basis of our concurrent host range, inclusion body, and serological data, it is likely that SCSV is in actuality the previously described and currently accepted potyvirus species BiMoV, for which no previous sequence data existed. As part of a comprehensive viral disease management plan, it is recommended that TSA plants growing in and around lettuce-production areas be controlled along with other weed hosts of this virus. References: (1) A. A. Brunt et al., eds. Plant Viruses Online: Descriptions and Lists from the VIDE Database. Version: 20 at http://biology.anu.edu.au/Groups/MES/vide/ , 1996. (2) A. Gibbs and A. J. Mackenzie. Virol. Methods 63:9, 1997. (3) R. J. McGovern et al. Int. J. Pest Manag. 40:270, 1994. (4) J. J. Mullahey et al. Weed Technol. 7:783, 1993.

scholarly journals Duplex-RT-PCR assay for the simultaneous detection and discrimination of Brome mosaic virus and Cocksfoot mottle virus in cereal plants

2021 ◽  
Author(s):  
Katarzyna Trzmiel
Keyword(s):  
Mosaic Virus ◽  
Simultaneous Detection ◽  
Brome Mosaic Virus ◽  
Mottle Virus ◽  
Grass Species ◽  
Replicase Gene ◽  
Rt Pcr ◽  
Pcr Products ◽  
Cereal Plants ◽  
Cocksfoot Mottle Virus

AbstractBrome mosaic virus (BMV) and cocksfoot mottle virus (CfMV) are pathogens of grass species including all economically important cereals. Both viruses have been identified in Poland therefore they create a potential risk to cereal crops. In this study, a duplex—reverse transcription—polymerase chain reaction (duplex-RT-PCR) was developed and optimized for simultaneous detection and differentiation of BMV and CfMV as well as for confirmation of their co-infection. Selected primers CfMVdiag-F/CfMVdiag-R and BMV2-F/BMV2-R amplified 390 bp and 798 bp RT-PCR products within coat protein (CP) region of CfMV and replicase gene of BMV, respectively. Duplex-RT-PCR was successfully applied for the detection of CfMV-P1 and different Polish BMV isolates. Moreover, one sample was found to be co-infected with BMV-ML1 and CfMV-ML1 isolates. The specificity of generated RT-PCR products was verified by sequencing. Duplex-RT-PCR, like conventional RT-PCR, was able to detect two viruses occurring in plant tissues in very low concentration (as low as 4.5 pg/µL of total RNA). In contrast to existing methods, newly developed technique offers a significant time and cost-saving advantage. In conclusion, duplex-RT-PCR is a useful tool which can be implemented by phytosanitary services to rapid detection and differentiation of BMV and CfMV.

scholarly journals Isolado do vírus do mosaico do pepino obtido de bananeira no Estado de São Paulo pertence ao subgrupo Ia

2001 ◽  
Vol 26 (1) ◽  
pp. 53-59 ◽  
Author(s):  
MARCELO EIRAS ◽  
ADDOLORATA COLARICCIO ◽  
ALEXANDRE L.R. CHAVES
Keyword(s):  
Mosaic Virus ◽  
Vigna Unguiculata ◽  
Sao Paulo ◽  
São Paulo ◽  
Rt Pcr ◽  
Nicotiana Glutinosa ◽  
Musa Sp ◽  
De Se ◽  
Pcr Rflp ◽  
Das Elisa

Em 1996, foi feita a caracterização parcial de um isolado do vírus do mosaico do pepino (Cucumis mosaic virus, CMV) obtido de bananeira (Musa sp.) proveniente do município de Miracatu, SP. Com o objetivo de se determinar o subgrupo do isolado de CMV, recorreu-se às técnicas de ELISA, RT-PCR, RFLP e seqüenciamento de fragmentos de RNA genômico. Amostras de folhas infetadas, desidratadas com cloreto de cálcio e armazenadas à -20 °C desde 1994 na viroteca do Laboratório de Fitovirologia e Fisiopatologia, foram inoculadas em plantas de Nicotiana glutinosa. Dez dias após a inoculação, folhas apresentando mosaico foram utilizadas para DAS-ELISA e extração de RNAs totais. Em ELISA, houve reação apenas contra o anti-soro específico para CMV subgrupo I. Através de RT-PCR com primers desenhados para anelar em regiões conservadas da porção terminal 3' do gene da capa protéica, foi amplificado um fragmento de DNA com 486 pares de bases. O produto obtido via RT-PCR foi submetido à digestão com as enzimas EcoRI, HindIII, BamHI e MspI, obtendo-se um padrão de restrição esperado para o subgrupo I. Estes resultados foram confirmados através do seqüenciamento do produto de PCR, o qual apresentou homologia de 96% a 98% com os isolados do CMV pertencentes ao subgrupo I. Pelos sintomas observados na hospedeira diferencial Vigna unguiculata, o isolado foi confirmado como sendo do subgrupo Ia.

scholarly journals Variabilidade genética de Sugarcane mosaic virus, causando mosaico em milho no Brasil

2011 ◽  
Vol 46 (4) ◽  
pp. 362-369 ◽  
Author(s):  
Marcos Cesar Gonçalves ◽  
Diogo Manzano Galdeano ◽  
Ivan de Godoy Maia ◽  
César Martins Chagas
Keyword(s):  
Mosaic Virus ◽  
Sao Paulo ◽  
Sugarcane Mosaic Virus ◽  
Maize Dwarf Mosaic Virus ◽  
São Paulo ◽  
Rt Pcr ◽  
Das Elisa

O objetivo deste trabalho foi caracterizar biológica e molecularmente três isolados de Sugarcane mosaic virus (SCMV) de lavouras de milho, analisá-los filogeneticamente e discriminar polimorfismos do genoma. Plantas com sintomas de mosaico e nanismo foram coletadas em lavouras de milho, no Estado de São Paulo e no Município de Rio Verde, GO, e seus extratos foliares foram inoculados em plantas indicadoras e submetidos à análise sorológica com antissoros contra o SCMV, contra o Maize dwarf mosaic virus (MDMV) e contra o Johnsongrass mosaic virus (JGMV). Mudas de sorgo 'Rio' e 'TX 2786' apresentaram sintomas de mosaico após a inoculação dos três isolados, e o DAS-ELISA confirmou a infecção pelo SCMV. O RNA total foi extraído e usado para amplificação por transcriptase reversa seguida de reação em cadeia de polimerase (RT-PCR). Fragmentos específicos foram amplificados, submetidos à análise por polimorfismo de comprimento de fragmento de restrição (RFLP) e sequenciados. Foi possível discriminar os genótipos de SCMV isolados de milho de outros isolados brasileiros do vírus. Alinhamentos múltiplos e análises dos perfis filogenéticos corroboram esses dados e mostram diversidade nas sequências de nucleotídeos que codificam para a proteína capsidial, o que explica o agrupamento separado desses isolados e sugere sua classificação como estirpes distintas, em lugar de simples isolados geográficos.

scholarly journals Detection and molecular characterization of Pepper mild mottle virus in Serbia

Genetika ◽  
2015 ◽  
Vol 47 (2) ◽  
pp. 651-663 ◽  
Author(s):  
Dragana Milosevic ◽  
Ivana Stankovic ◽  
Aleksandra Bulajic ◽  
Maja Ignjatov ◽  
Zorica Nikolic ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Molecular Characterization ◽  
Great Influence ◽  
Alfalfa Mosaic Virus ◽  
Resistance Breeding ◽  
Elisa Test ◽  
Mottle Virus ◽  
Pepper Mild Mottle Virus ◽  
Das Elisa

During 2009 and 2010, a survey was conducted in pepper crops to detect the possible presence of Pepper mild mottle virus (PMMoV) in Serbia. A total of 239 pepper samples from 39 crops at 26 localities were collected and analyzed for the presence of PMMoV, Cucumber mosaic virus (CMV), Potato virus Y (PVY), and Alfalfa mosaic virus (AMV), using DAS-ELISA test. Although it was detected in a small percentage, PMMoV could pose a threat to pepper production in Serbia due to its rapid seed-borne spread. Presence of PMMoV was confirmed by serological and biological detection, followed by conventional reverse transcription RT-PCR, using primers specific for the RNA-dependent RNA polymerase (RdRp) and the coat protein (CP) genes. Molecular identification confirmed that the Serbian isolates belong to PMMoV pathotypes P1,2 which do not break the resistance gene L3. Reconstructed phylogenetic tree confirmed the allocation of the Serbian isolates together with the majority of PMMoV isolates which belong to pathotypes P1,2. This study represents the first serological and molecular characterization of PMMoV infection of pepper in Serbia, and provides important data on the population structure. The obtained data could have great influence on pepper production in Serbia as well as future pepper resistance breeding in the country.

scholarly journals KETAHANAN BEBERAPA KULTIVAR CABAI TERHADAP CUCUMBER MOSAIC VIRUS DAN CHILLI VEINAL MOTTLE VIRUS

2012 ◽  
Vol 7 (2) ◽  
pp. 130-139
Author(s):  
Muhammad Taufik ◽  
Sri Hendrastuti Hidayat ◽  
Sriani Sujiprihati ◽  
Gede Suastika ◽  
Sientje Mandang Sumaraw
Keyword(s):  
Virus Infection ◽  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Yield Loss ◽  
Disease Incidence ◽  
Growth And Yield ◽  
Mottle Virus ◽  
Tolerance Response ◽  
Chilli Veinal Mottle Virus ◽  
Das Elisa

Resistance Evaluation of Chillipepper Cultivars for Cucumber Mosaic Virus and Chilli Veinal Mottle Virus.  The use of resistance culivars is an important strategy for management of virus infection in chillipepper. A research was undergone to study the effect of single and mix infection of CMV and ChiVMV on the disease incidence and on the growth and yield of nine chillipepper cultivars, i.e. Cilibangi 4, Cilibangi 5, Cilibangi 6, Helem, Jatilaba, Tit Bulat, Tit Segitiga, Tit Super and Tampar. Mechanical inoculation was conducted to transmit the virus. Infection of the virus was then confirmed with DAS-ELISA.  In general, inoculated chillipepper cultivars developed similar symptoms, i.e. mosaic type for CMV and mottle type for ChiVMV.  More severe symptom was not always observed from mix infection of CMV and ChiVMV. Disease incidence occurred in the range of 16.67 – 86.0% and this caused 18.3 – 98.6% yield loss.  Based on symptom expression, ELISA result, and reduction on yield, it can be concluded that all chillipepper cultivars used in this study could not hold up the virus infection. However, several cultivars showed tolerance response :  Jatilaba, Tit Super, and Tampar for CMV; Cilibangi 4 for ChiVMV; Tit Super for mix infection; and Cilibangi 5 for CMV, ChiVMV, and mix infection.  Further evaluation and investigation involving different chillipepper cultivars should be conducted.

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 Lamium maculatum is a Natural Host for Cucumber mosaic virus

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 150-150 ◽  
Author(s):  
R. Bešta-Gajević ◽  
A. Jerković-Mujkić ◽  
S. Pilić ◽  
I. Stanković ◽  
A. Vučurović ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Impatiens Necrotic Spot Virus ◽  
Natural Occurrence ◽  
Rt Pcr ◽  
Natural Ecosystems ◽  
New Host ◽  
Total Rna ◽  
Negative Controls ◽  
Das Elisa

Lamium maculatum L. (spotted dead-nettle) is a flowering perennial ornamental that is commonly grown as a landscape plant for an effective ground cover. In June 2010, severe mosaic accompanied by reddish brown necrosis and leaf deformation was noticed on 80% of L. maculatum growing in shade under trees and shrubs in Sarajevo (Bosnia and Herzegovina). Leaves from 10 symptomatic L. maculatum plants were sampled and analyzed by double-antibody sandwich (DAS)-ELISA using commercial diagnostic kits (Bioreba AG, Reinach, Switzerland) against Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV), and Impatiens necrotic spot virus (INSV), the most important viral pathogens of ornamental plants (1,2). Commercial positive and negative controls and extracts from healthy L. maculatum leaves were included in each assay. All samples tested negative for TSWV and INSV and positive for CMV. The virus was mechanically transmitted to test plants and young virus-free plants of L. maculatum using 0.01 M phosphate buffer (pH 7). The virus caused chlorotic local lesions on Chenopodium quinoa, while systemic mosaic was observed on Capsicum annuum ‘Rotund,’ Nicotiana rustica, N. glutinosa, N. tabacum ‘White Burley,’ and Phaseolus vulgaris ‘Top Crop.’ The virus was transmitted mechanically to L. maculatum and induced symptoms resembling those observed on the source plants. Inoculated plants were assayed by DAS-ELISA and all five inoculated plants of each species tested positive for CMV. The presence of CMV in L. maculatum as well as mechanically infected N. glutinosa plants was further confirmed by RT-PCR. Total RNA from symptomatic leaves was isolated using RNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and RT-PCR was performed with the One-Step RT-PCR Kit (Qiagen) following the manufacturer's instructions. The primer pair, CMVAu1u/CMVAu2d, that amplifies the entire coat protein (CP) gene and part of 3′- and 5′-UTRs was used for both amplification and sequencing (4). Total RNA obtained from the Serbian CMV isolate from pumpkin (GenBank Accession No. HM065510) and a healthy L. maculatum plant were used as positive and negative controls, respectively. All naturally and mechanically infected plants as well as the positive control yielded an amplicon of the expected size (850 bp). No amplicon was observed in the healthy control. The amplified product derived from isolate 3-Lam was purified (QIAquick PCR Purification Kit, Qiagen), directly sequenced in both directions and deposited in GenBank (JX436358). Sequence analysis of the CP open reading frame (657 nt), conducted with MEGA5 software, revealed that the isolate 3-Lam showed the highest nucleotide identity of 99.4% (99.1% amino acid identity) with CMV isolates from Serbia, Australia, and the USA (GQ340670, U22821, and U20668, respectively). To our knowledge, this is the first report of the natural occurrence of CMV on L. maculatum worldwide and it adds a new host to over 1,241 species (101 plant families) infected by this virus (3). This is also an important discovery for the ornamental industry since L. maculatum is commonly grown together with other ornamental hosts of CMV in nurseries and the urban environment as well as in natural ecosystems. References: (1) Y. K. Chen et al. Arch. Virol. 146:1631, 2001. (2) M. L. Daughtrey et al. Plant Dis. 81:1220, 1997. (3) M. Jacquemond. Adv. Virus Res. 84:439, 2012. (4) I. Stankovic et al. Acta Virol. 55:337, 2011.

scholarly journals Occurrence of Stone Fruit Viruses in Plum Orchards in Latvia

2013 ◽  
Vol 67 (2) ◽  
pp. 116-123
Author(s):  
Alina Gospodaryk ◽  
Inga Moročko-Bičevska ◽  
Neda Pūpola ◽  
Anna Kāle
Keyword(s):  
Mosaic Virus ◽  
Large Scale ◽  
Ringspot Virus ◽  
Stone Fruit ◽  
Detection Methods ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Detection Rates ◽  
Arabis Mosaic Virus ◽  
Das Elisa

To evaluate the occurrence of nine viruses infecting Prunus a large-scale survey and sampling in Latvian plum orchards was carried out. Occurrence of Apple mosaic virus (ApMV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), Apple chlorotic leaf spot virus (ACLSV), and Plum pox virus (PPV) was investigated by RT-PCR and DAS ELISA detection methods. The detection rates of both methods were compared. Screening of occurrence of Strawberry latent ringspot virus (SLRSV), Arabis mosaic virus (ArMV), Tomato ringspot virus (ToRSV) and Petunia asteroid mosaic virus (PeAMV) was performed by DAS-ELISA. In total, 38% of the tested trees by RT-PCR were infected at least with one of the analysed viruses. Among those 30.7% were infected with PNRSV and 16.4% with PDV, while ApMV, ACLSV and PPV were detected in few samples. The most widespread mixed infection was the combination of PDV+PNRSV. Observed symptoms characteristic for PPV were confirmed with RT-PCR and D strain was detected. Comparative analyses showed that detection rates by RT-PCR and DAS ELISA in plums depended on the particular virus tested. The results obtained in this study revealed that commonly grown plum cultivars in Latvia are infected with economically important stone fruit viruses and highlight the need to implement a programme to produce and propagate virus-free planting material.

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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