scholarly journals First Report of Sweet potato chlorotic stunt virus and Sweet potato feathery mottle virus Infecting Sweet Potato in Spain

Plant Disease ◽  
2004 ◽  
Vol 88 (4) ◽  
pp. 428-428 ◽  
Author(s):  
R. A. Valverde ◽  
G. Lozano ◽  
J. Navas-Castillo ◽  
A. Ramos ◽  
F. Valdés
Keyword(s):  
Nucleotide Sequence ◽  
Sweet Potato ◽  
Morning Glory ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Degenerate Primers ◽  
Potato Plants ◽  
Elisa Testing ◽  
Two Samples

Sweet potato chlorotic stunt virus (SPCSV), family Closteroviridae and Sweet potato feathery mottle virus (SPFMV), family Potyviridae are whitefly and aphid transmitted, respectively, which in double infections cause sweet potato virus disease (SPVD) that is a serious sweet potato (Ipomoea batatas Lam.) disease in Africa (2). During the past decade, sweet potato plants showing symptoms similar to SPVD have been observed in most areas of Spain. Nevertheless, not much information is available about the identity of the viruses infecting this crop in Spain. During the summer of 2002, sweet potato plants with foliar mosaic, stunting, leaf malformation, chlorosis, and ringspot symptoms were observed in several farms in Málaga (southern Spain) and Tenerife and Lanzarote (Canary Islands, Spain). Vine cuttings were collected from 21 symptomatic plants in Málaga and from eight plants on Lanzarote and six on Tenerife. Scions were grafted to the indicator hosts, Brazilian morning glory (I. setosa) and I. nil cv. Scarlett O'Hara. Three weeks after graft inoculations, all plants showed various degrees of mosaic, chlorosis, leaf malformation, and stunting. Four field collections (two from Málaga, one from Tenerife, and one from Lanzarote) with severe symptoms on I. setosa were selected for whitefly (Bemisia tabaci biotype Q) transmission experiments. Acquisition and transmission periods were 48 h. I. setosa was the acquisition host, and I. nil was the transmission host. For each isolate, groups of 10 whiteflies per I. nil plant were used. All I. nil plants used as transmission hosts with the four, field collections showed chlorosis and leaf malformation. Reverse-transcription polymerase chain reaction (RT-PCR) was performed on I. setosa (grafted with the four selected field collections) and I. nil plants (from the whitefly transmission experiments) with primers for the HSP70h gene of SPCSV. A 450-bp DNA fragment was obtained with all I. setosa and I. nil samples. Sequencing of the 450-bp DNA from two samples from Málaga yielded a nucleotide sequence with 98 to 99% similarity to the HSP70h gene of West African SPCSV isolates. Foliar samples from I. setosa, originally grafted with the 21 vine cuttings, were used for nitrocellulose membrane enzyme-linked immunosorbent assay (NCM-ELISA) testing with antiserum specific to SPFMV-RC (provided by J. Moyer, North Carolina State University, Raleigh). Positive control was sap extract from I. setosa that was infected with the common strain of SPFMV. Procedures for NCM-ELISA were as described (4). NCM-ELISA testing suggested that SPFMV was present in all samples. RT-PCR was conducted with degenerate primers POT1/POT2 (1). The nucleotide sequence that was amplified by these two primers spans part of the NIb protein and part of the coat protein gene of potyviruses. All samples yielded the expected 1.3-kb DNA. Sequencing of the RT-PCR products of two isolates from Malaga and sequence comparisons yielded nucleotide sequences with 97% similarity to two East African isolates (Nam 1 and Nam 3) of SPFMV (3). These results confirm the presence of SPCSV and SPFMV in sweet potato in Spain. References: (1) D. Colinet and J. Kummert. J. Virol. Methods 45:149, 1993. (2) R. W. Gibson et al. Plant Pathol. 47:95, 1998. (3) J. F. Kreuze et al. Arch. Virol. 145:567, 2000. (4) E. R. Souto et al. Plant Dis. 87:1226, 2003.

scholarly journals First Report of Cherry virus A and Little cherry virus-1 in Poland

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 909-909 ◽  
Author(s):  
B. Komorowska ◽  
M. Cieślińska
Keyword(s):  
Nucleotide Sequence ◽  
Coat Protein ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Germplasm Collection ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
First Report ◽  
Primer Sets

Cherry virus A (CVA), a member of the genus Capillovirus, has been reported in sweet cherry in Germany, Canada, and Great Britain. No data are available on the effects of CVA on fruit quality and yield of infected trees. Little cherry disease (LChD) occurs in most cherry growing areas of the world. Symptoms on sensitive cultivars include discolored fruit that remain small, pointed in shape, and tasteless. Three Closterovirus spp. associated with LChD have been described (Little cherry virus-1 [LChV-1], LChV-2, and LChV-3). Diseased local and commercial cultivars of sour cherry trees were found in a Prunus sp. germplasm collection and orchards in Poland during the 2003 growing season. The foliar symptoms included irregular, chlorotic mottling, distortion, and premature falling of leaves. Some of the diseased trees developed rosette as a result of decreased growth and shortened internodes. Severely infected branches exhibited dieback symptoms. Because the symptoms were suggestive of a possible virus infection, leaf samples were collected from 38 trees and assayed for Prune dwarf virus and Prunus necrotic ringspot virus using double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). RNA extracted from leaves was used in a reverse transcription-polymerase chain reaction (RT-PCR) with the One-Step RT-PCR with Platinum Taq (Invitrogen Life Technologies) and primer sets specific for CVA (1), LChV-1 (3), and LChV-2 (3). The RNA samples were also tested using RT-PCR for detection of Cherry mottle leaf virus (CMLV), Cherry necrotic rusty mottle virus (CNRMV), and Cherry green ring mottle virus (CGRMV) with specific primer sets (2). Amplification of a 397-bp coat protein gene product confirmed infection of 15 trees with CVA. A 419-bp fragment corresponding to the coat protein gene of LChV-1 was amplified from cv. Gisela rootstock and local cv. WVIII/1. To confirm RT-PCR results, CVA amplification products from local cv. WX/5 and LChV-1 from cvs. Gisela and WVIII/1 were cloned in bacterial vector pCR 2.1-TOPO and then sequenced. The sequences were analyzed with the Lasergene (DNASTAR, Madison, WI) computer program. The alignment indicated that the nucleotide sequence of cv. WX/5 was closely related to the published sequences of CVA (Genbank Accession No. NC_003689) and had an 89% homology to the corresponding region. The nucleotide sequence similarity between the 419-bp fragment obtained from cvs. Gisela and WVIII/1 was 87% and 91%, respectively, compared with the reference isolate of LChV-1 (Genbank Accession No. NC_001836). The sampled trees tested negative for LChV-2, CGRMV, CMLV, and CNRMV using RT-PCR. Some trees tested positive for PNRSV and PDV. To our knowledge, this is the first report of CVA and LChV-1 in Poland. References: (1) D. James and W. Jelkmann. Acta Hortic. 472:299, 1998. (2) M. E. Rott and W. Jelkmann. Eur. J. Plant Pathol. 107:411,2001. (3) M. E. Rott and W. Jelkmann. Phytopathology. 91:61, 2001.

scholarly journals First Report of Pepper mottle virus in Tomato

Plant Disease ◽  
2002 ◽  
Vol 86 (2) ◽  
pp. 186-186 ◽  
Author(s):  
J. Th. J. Verhoeven ◽  
T. M. Willemen ◽  
J. W. Roenhorst
Keyword(s):  
Nucleotide Sequence ◽  
Lycopersicon Esculentum ◽  
Potato Virus ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Degenerate Primers ◽  
Pepper Mottle Virus ◽  
First Report ◽  
Transmission Electron ◽  
Nontranslated Region

In 2000, a breeding company submitted a tomato (Lycopersicon esculentum) sample from Guatemala for diagnosis. The plants showed necrotic lesions on leaves surrounded by some chlorosis, necrotic streaks on stems, and large superficial necrotic lesions on fruits. By mechanical inoculation of plant sap to different plant species, symptoms appeared in Capsicum annuum ‘Westlandse Grote Zoete’, Lycopersicon esculentum ‘Money-maker’, Nicotiana benthamiana, N. bigelovii, N. glutinosa, N. hesperis-67A, N. occidentalis-P1, N. tabacum ‘White Burley’, and Physalis floridana. Systemically infected leaves from N. occidentalis-P1 were used for all further experiments. Leaf dip preparations were analyzed by transmission electron microscopy and revealed the presence of filamentous virus particles typical of a potyvirus. Double-antibody sandwich enzyme-linked immunosorbent assay tests for Potato virus A, V, and Y, Tobacco etch virus, and Wild potato mosaic virus were negative. An antiserum (PepMoV/DSMZ As 0186) to Pepper mottle virus (PepMoV), however, gave a positive reaction. To obtain further evidence for the presence of this virus, the nucleotide sequence of the complete 3′ nontranslated region (3NTR) and the 3′ terminal part of the coat protein gene (3CPG) was determined using the set of degenerate primers P9502/CPUP (1). The obtained nucleotide sequence (approximately 700 bp) was deposited in GenBank under Accession No. AF440801. It showed 93 to 94% 3NTR and 90 to 93% 3CPG homology with the three sequences of PepMoV from pepper already present in GenBank. The two viruses with the next closest nucleotide sequence homology were Potato virus V and Potato virus Y showing up to 80 and 75% homology with the 3CPG and up to 53 and 48% homology with the 3NTR, respectively. Based on these results, we concluded that the virus isolated from the symptomatic tomato plants was PepMoV. Because of the relatively low homologies with the pepper isolates of PepMoV, this tomato isolate might be a separate strain of the virus. To our knowledge, this is the first report of the occurrence of PepMoV in tomato. Reference: (1) R. A. A. van der Vlugt et al. Phytopathology 89:148, 1999.

scholarly journals First report of groundnut ringspot virus infecting Zinnia sp. in Brazil

Plant Disease ◽  
2021 ◽  
Author(s):  
Felipe Franco de Oliveira ◽  
Gabriel Madoglio Favara ◽  
Camila Geovana Ferro ◽  
Heron Delgado Kraide ◽  
Eike Yudi Nishimura Carmo ◽  
...  
Keyword(s):  
Nucleotide Sequence ◽  
Nucleotide Sequences ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Concentric Ring ◽  
Leaf Extracts ◽  
Total Rna ◽  
First Report ◽  
Two Samples

Zinnia sp. is a genus belonging to Asteraceae family, originated in Mexico and adapted to a warm-hot climate (Hemmati and Mehrnoosh, 2017). Several types of zinnias with different flower color and forms are cultivated in Brazil (Min et al., 2020 and Souza Jr. et al., 2020). Characteristic symptoms of infection caused by orthotospovirus, including chlorotic spots and concentric rings on the leaves, were observed in two plants of Zinnia sp. of a florist located in the city of Piracicaba, State of São Paulo, Brazil. Orthotospovirus-like particles were observed by transmission electron microscope in leaf extracts from both plants, stained negatively with 1% uranyl acetate. By analyzing ultrathin sections of infected leaf tissues, particles of 80-100 nm in diameter were found in the lumen of the endoplasmic reticulum and nucleocapsid aggregates in the cytoplasm. Total RNA extracted separately from the leaves of both samples, using the Purelink Viral DNA / RNA kit (Thermo Fisher Scientific), was used to detect the virus by reverse transcription polymerase chain reaction (RT-PCR), using the universal primers for orthotospovirus BR60, complementary to the 3’ end of the non-translated region of the S RNA (position 1 to 15 nt), and BR65, matching the nucleocapsid gene (N) (position 433 to 453 nt), generating and amplicon of 453 nt (Eiras et al., 2001). Amplicons of the expected size were obtained for the two samples. An amplicon was purified with the Wizard SV Gel and PCR Clean-Up System kit (Promega) and sequenced in both directions at Macrogen Inc (South Korea). The nucleotide sequence (GenBank MW629018) showed 99.29-99.76% identity with nucleotide sequences of the orthotospovirus groundnut ringspot virus (GRSV) isolates (GenBank MH686229 and KY400110). Leaf extracts from symptomatic plants were also analyzed by plate-trapped antigen-enzyme-linked immunosorbent assay (PTA-ELISA), using polyclonal antiserum produced against the GRSV nucleocapsid protein (Esquivel et al., 2019). The absorbance values obtained for the extracts of the two symptomatic plants of Zinnia sp. (1.3 and 1.7) were twice as high as the value obtained for the healthy plant extract (0.5). Leaf extract of symptomatic Zinnia sp. was inoculated mechanically onto leaves of healthy plants of Zinnia sp., Capsicum annuum cv. Dara, Cucumis sativus, Cucurbita pepo cv. Caserta, Chenopodium amaranticolor, Datura stramonium, Nicotiana tabacum cv. Turkish and Solanum lycopersicum cv. Compack. At 5 days post inoculation (dpi), inoculated leaves of D. stramonium reacted with local lesions, and at 9 dpi, newly developed leaves of inoculated S. lycopersicum plants showed necrotic spot and concentric ring symptoms, whereas C. annuum exhibited concentric rings at 10 dpi. Inoculated zinnia plants showed systemic chlorotic spot and concentric ring symptoms at 20 dpi, indistinguishable from those observed under natural infection. The other inoculated plant species were not symptomatic, nor the virus was detected. PTA-ELISA and RT-PCR confirmed infection with GRSV in symptomatic plants. The amplicons generated by RT-PCR of total RNA extracted from an experimentally infected plant of C. annuum and D. stramonium, and two plants of Zinnia sp. were sent for nucleotide sequencing. The obtained nucleotide sequences (MW629019, MW629020, MW629021, MW629022) shares 100% identity with the nucleotide sequence corresponding to the original GRSV isolate (MW629018) identified in Zinnia sp. This is the first report of the natural occurrence of GRSV in Zinnia sp. in Brazil. Studies on incidence and damage are needed to recommend alternatives for management.

scholarly journals Incidence and distribution of Sweetpotato viruses and their implication on sweetpotato seed system in Malawi

2021 ◽  
Author(s):  
Willard Mbewe ◽  
Andrew Mtonga ◽  
Margret Chiipanthenga ◽  
Kennedy Masamba ◽  
Gloria Chitedze ◽  
...  
Keyword(s):  
Sweet Potato ◽  
Virus Disease ◽  
Disease Incidence ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Foliar Symptoms ◽  
Membrane Enzyme ◽  
Sweet Potato Virus ◽  
Seed System ◽  
Clean Seed

AbstractA survey was carried out in 19 districts to investigate the prevalence and distribution of sweetpotato virus disease (SPVD) and its implication on the sustainability of clean seed system in Malawi. A total of 166 leaf samples were collected and tested for the presence of 8 viruses using nitrocellulose membrane enzyme-linked immunosorbent assay (NCM-ELISA). SPVD foliar symptoms were observed in 68.42% of the surveyed districts. There were significant variations in disease incidence and severity (p < 0.001) among districts, with the highest incidence in Mulanje (28.34%). Average SPVD severity score was 3.05. NCM-ELISA detected sweet potato feathery mottle virus (SPFMV, 30.54%), sweet potato mild mottle virus (SPMMV, 31.14%), sweet potato mild speckling virus (SPMSV, 16.17%), sweet potato C-6 virus (SPC6V, 13.77%), sweet potato chlorotic stunt virus (SPCSV, 22.16%), sweet potato collusive virus (SPCV, 30.54%), sweet potato virus G (SPVG, 11.38%), cucumber mosaic virus (CMV, 7.78%) either in single or mixed infections. Data from this study indicate a significant SPVD occurrence in the country, and the consequence implications towards national sweetpotato seed system.

scholarly journals First Report of Mosaic Disease Caused by Colombian datura virus on Solanum lycopersicum Plants Commercially Cultivated in Japan

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 698-698 ◽  
Author(s):  
Y. Tomitaka ◽  
T. Usugi ◽  
R. Kozuka ◽  
S. Tsuda
Keyword(s):  
Nucleotide Sequence ◽  
Solanum Lycopersicum ◽  
Mosaic Disease ◽  
Causal Agent ◽  
Cultivated Plants ◽  
Rt Pcr ◽  
Degenerate Primers ◽  
Specific Primers ◽  
Tomato Plants ◽  
First Report

In 2009, some commercially grown tomato (Solanum lycopersicum) plants in Chiba Prefecture, Japan, exhibited mosaic symptoms. Ten plants from a total of about 72,000 cultivated plants in the greenhouses showed such symptoms. To identify the causal agent, sap from leaves of the diseased plants was inoculated into Chenopodium quinoa and Nicotiana benthamiana plants. Local necrotic lesions appeared on inoculated leaves of C. quinoa, but no systemic infection was observed. Systemic mosaic symptoms were observed on the N. benthamiana plants inoculated. Single local lesion isolation was performed three times using C. quinoa to obtain a reference isolate for further characterization. N. benthamiana was used for propagation of the isolate. Sap from infected leaves of N. benthamiana was mechanically inoculated into three individual S. lycopersicum cv. Momotaro. Symptoms appearing on inoculated tomatoes were indistinguishable from those of diseased tomato plants found initially in the greenhouse. Flexuous, filamentous particles, ~750 nm long, were observed by electron microscopy in the sap of the tomato plants inoculated with the isolate, indicating that the infecting virus may belong to the family Potyviridae. To determine genomic sequence of the virus, RT-PCR was performed. Total RNA was extracted from the tomato leaves experimentally infected with the isolate using an RNeasy Plant Mini kit (QIAGEN, Hilden, Germany). RT-PCR was performed by using a set of universal, degenerate primers for Potyviruses as previously reported (2). Amplicons (~1,500 bp) generated by RT-PCR were extracted from the gels using the QIAquick Gel Extraction kit (QIAGEN) and cloned into pCR-BluntII TOPO (Invitrogen, San Diego, CA). DNA sequences of three individual clones were determined using a combination of plasmid and virus-specific primers, showing that identity among three clones was 99.8%. A consensus nucleotide sequence of the isolate was deposited in GenBank (AB823816). BLASTn analysis of the nucleotide sequence determined showed 99% identity with a partial sequence in the NIb/coat protein (CP) region of Colombian datura virus (CDV) tobacco isolate (JQ801448). Comparison of the amino acid sequence predicted for the CP with previously reported sequences for CDV (AY621656, AJ237923, EU571230, AM113759, AM113754, and AM113761) showed 97 to 100% identity range. Subsequently, CDV infection in both the original and experimentally inoculated plants was confirmed by RT-PCR using CDV-specific primers (CDVv and CDVvc; [1]), and, hence, the causal agent of the tomato disease observed in greenhouse tomatoes was proved to be CDV. The first case of CDV on tomato was reported in Netherlands (3), indicating that CDV was transmitted by aphids from CDV-infected Brugmansia plants cultivated in the same greenhouse. We carefully investigated whether Brugmansia plants naturally grew around the greenhouses, but we could not find them inside or in proximity to the greenhouses. Therefore, sources of CDV inoculum in Japan are still unclear. This is the first report of a mosaic disease caused by CDV on commercially cultivated S. lycopersicum in Japan. References: (1) D. O. Chellemi et al. Plant Dis. 95:755, 2011. (2) J. Chen et al. Arch. Virol. 146:757, 2001. (3) J. Th. J. Verhoeven et al. Eur. J. Plant. Pathol. 102:895, 1996.

scholarly journals Epidemiological Survey and Screening Strategy for Dengue Virus in Blood Donors from Yunnan Province

2020 ◽  
Author(s):  
LING LI ◽  
YING LI ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Prevalence Rate ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Igm Antibodies ◽  
Blood Center ◽  
Nonstructural Protein 1 ◽  
Elisa Testing

Abstract BACKGROUND Dengue virus (DENV) can be transmitted through blood transfusion. DENV was not screened regularly in Xishuangbanna Blood Center. This study was conducted in Xishuangbanna Blood Center with an attempt to develop DENV screening strategies in one of China’s high-incidence areas.METHODS Blood samples were collected randomly between June 2019 and August 2019. These samples were first screened for dengue IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). All reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real time polymerase-chain-reaction (RT-PCR) assay. After RT-PCR assay, these samples were further tested for soluble nonstructural protein 1 (NS1) using colloidal gold method. The demographic data of DENV positive donors were collected.RESULTS A total of 2,254 donor samples were collected and tested for dengue IgG and IgM antibodies by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-IgG and/or anti-IgM reactive, with a serological prevalence rate of 26.53%. Among all the donor samples, 26 were RT-PCR positive and/or NS1 positive. Moreover, there were significant differences in the prevalence rate of DENV in terms of occupation (P=0.001), education(P<0.001) and ethnicity (P=0.026). CONCLUSION The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides the first-hand data about the prevalence of DENV and allows development of a screening strategy for clinical use.

scholarly journals Effects of sweet potato feathery mottle virus, sweet potato chlorotic stunt virus and their co-infection on sweet potato yield in Western Burkina Faso

2019 ◽  
Vol 4 (1) ◽  
pp. 758-766
Author(s):  
E. B. Tibiri ◽  
K. Somé ◽  
J. S. Pita ◽  
F. Tiendrébéogo ◽  
M. Bangratz ◽  
...  
Keyword(s):  
Burkina Faso ◽  
Sweet Potato ◽  
Virus Disease ◽  
Block Design ◽  
Potato Yield ◽  
Mottle Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Yield Reduction ◽  
Potato Varieties ◽  
Western Burkina Faso

AbstractTo determine the effects of sweet potato feathery mottle virus (SPFMV), Sweet potato chlorotic stunt virus (SPCSV) and their co-infection on sweet potato yield, twelve sweet potato varieties were assessed in a hotspot area in Western Burkina Faso. The experiment was carried out in a randomized complete-block design with the twelve varieties in three replications. Data were collected on plant growth parameters, plant virus symptoms and yield parameters. Additional testing for selected sweet potato viruses was done using a nitrocellulose membrane enzyme-linked immunosorbent assay (NCM-ELISA) and RT-PCR. SPFMV and SPCSV were the viruses detected in this study. Varieties Djakani and Ligri were virus-free and had the highest average yields out of twelve sweet potato varieties assessed. Field monitoring indicated that 58% of plants were found to be virus-infected. The results suggest that severe symptoms were associated with sweet potato virus disease (SPVD) and yield reduction. However, the interaction of SPCSV with other viruses, which may result in synergistic negative effects on sweet potato yield and quality, needs further research.

scholarly journals First Report of Potato mop-top virus on Potato from the United States

Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 872-872 ◽  
Author(s):  
D. H. Lambert ◽  
L. Levy ◽  
V. A. Mavrodieva ◽  
S. B. Johnson ◽  
M. J. Babcock ◽  
...  
Keyword(s):  
Triple Gene Block ◽  
The United States ◽  
Russet Burbank ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coding Region ◽  
Rt Pcr ◽  
Protein Coding ◽  
Gene Block ◽  
Elisa Testing ◽  
Potato Mop Top Virus

Potato mop-top virus (PMTV) is a tripartite pomovirus vectored by the powdery scab plasmodiophoromycete Spongospora subterranea pv. subterranea (1). PMTV occurs on potato (Solanum tuberosum) in Europe, the Andes, Asia, and Canada. Internal necrotic arc and fleck tuber symptoms (“spraing”) may reduce commercial acceptance of some cultivars (3). PMTV symptoms were discovered in ‘Shepody’ tubers at the Aroostook Research Farm, Presque Isle, ME in May 2002 and subsequently in ‘Russet Burbank’ tubers in commercial storage from the 2001 Maine crop. Symptomatic tubers exhibited single or multiple concentric necrotic arcs that were partial or complete, but exhibited no distinct external symptoms. The presence of PMTV in eight ‘Shepody’ tubers was indicated by positive enzyme-linked immunosorbent assay (ELISA; Adgen, Ltd., Auchincruive, Ayr, Scotland) and confirmed by reverse transcription polymerase chain reaction (RT-PCR). ‘Russet Burbank’ potatoes were visually diagnosed, and the corresponding halves of 128 symptomatic tubers were forwarded to the University of Maine and APHIS (Beltsville, MD). Of these, ELISA readings in Maine were strongly positive (>3 × background) for 88, ambiguous (1.5-3 × background) for 13, and negative for 27. Subsamples from these three categories were positive by PCR in 17 of 17, 9 of 9, and 12 of 14 cases, respectively. A similar rating, positive or ambiguous, in ELISA testing was identical for all but one case at Beltsville. Confirmation of PMTV required PCR testing, resulting in a characteristic PCR product of 401 bp that was generated from the coat protein coding region on RNA 2 (2) using the primer pair PMTV 1 5′-GCAGCCGTCGAGAATAGATA-3′ (RNA nucleotides 316–335) and PMTV 4 5′-GCGAGTTGATGTGCC ACATT-3′ (complementary to RNA 2 nucleotides 716–697). An immunocapture RT-PCR using this primer set and the coating antibody from the Adgen ELISA kit was also successful in detecting PMTV. In separate reactions, a second product of 646 bp was generated from the triple gene block on RNA 3 (4) using the primer pair PMTV 5 5′-GGTGAACACGAGGACAAGGT-3′ (RNA 3 nucleotides 1417–1436) and PMTV 7 5′-AACAGTCCGGTCTTGTGAAC-3′ (complementary to RNA 3 nucleotides 2063–2044). The sequence of these products was 98 to 100% identical to PMTV published sequences. The discovery of this virus will result in adjustments to U.S. and Canadian seed potato certification standards and symptom characterization for common North American cultivars. References: (1) R. A. C. Jones and B. D. Harrison. Ann. Appl. Biol 63:1, 1969. (2) S. Kashiwazak et al. Virology 206:701, 1995. (3) M. Sandgren et al. Am. J. Potato Res. 79:205, 2002. (4) K. P. Scott et al. J. Gen. Virol.75:3561, 1994.

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.

Sign in / Sign up

Export Citation Format

Share Document