scholarly journals A New Geminivirus Associated with Tomato in the State of São Paulo, Brazil

Plant Disease ◽  
1997 ◽  
Vol 81 (4) ◽  
pp. 423-423 ◽  
Author(s):  
J. C. Faria ◽  
J. A. C. Souza-Dias ◽  
S. A. Slack ◽  
D. P. Maxwell

The apical growth of about 20% of young tomato plants in observed fields near Campinas, State of São Paulo, Brazil, had yellow streaking of veins. Leaf symptoms developed into patches of yellow mosaic and the leaves became wavy. The whitefly Bemisia tabaci Genn. transmitted a pathogen from the infected tomato plants to healthy tomato and potato plants, reproducing the original symptoms in tomato. The apical leaves of infected potatoes showed yellow or green mottle that developed into leaf distortion with yellow blotches, symptoms indistinguishable from potato-deforming mosaic disease (2). DNA was extracted from these tomato and potato plants (1). Using DNA from the infected tomato plant, polymerase chain reaction (PCR) with the degenerate primer pair PAC1v1978/ PAV1c715 (1), which amplifies part of the rep gene (AC1 ORF), the common region (CR), and part of the cp gene (AV1 ORF), and with the primer pair PBC1v2039/PBV1c800, which amplifies part of BC1 ORF, CR, and part of BV1 ORF, gave virus-specific DNA fragments of the sizes expected from a whitefly-transmitted geminivirus. These were cloned and the complete nucleotide (sequences for DNA-A (pToYA, GenBank accession no. U79998) and DNA-B (pToYB, GenBank accession no. U80042) fragments obtained. Nucleotide identity between the CRs (184 nucleotides) was 90%, strongly indicating that those fragments correspond to a bipartite subgroup III geminivirus. PCR with the DNA from infected potato gave the expected size fragment for DNA-A. The partial sequence of the rep gene was 100% identical to the homologous sequence from the PCR fragment from the infected tomato. A search in the GenBank, EMBL, DDBJ, and PDB databases, using the BLAST program, found no identical geminivirus. The highest identity for the CR was 75% to tomato mottle geminivirus-Florida (ToMoV) and 74% to bean golden mosaic virus-Brazil. For the rep gene, the highest identity was 73% to tomato yellow leaf curl virus-Israel, an Old World geminivi-rus, followed by 71% to tomato golden mosaic virus-Brazil (TGMV) and ToMoV. For the cp gene, the highest identity was 86% to TGMV, followed by 83% to squash leaf curl geminivirus. Therefore, we propose the name tomato yellow vein streak geminivirus (ToYVSV) for this distinct virus (2). References: (1) M. R. Rojas et al. Plant Dis. 77:340, 1993. (2) J. A. C. Souza-Dias et al. Summa Phytopathol. 22:57, 1996.

2018 ◽  
Vol 44 (3) ◽  
pp. 297-301
Author(s):  
Lígia M. L. Duarte ◽  
Maria Amelia V. Alexandre ◽  
Alexandre L. R. Chaves ◽  
Ricardo Harakava ◽  
Luiz G. B. Alvim ◽  
...  

2001 ◽  
Vol 26 (1) ◽  
pp. 53-59 ◽  
Author(s):  
MARCELO EIRAS ◽  
ADDOLORATA COLARICCIO ◽  
ALEXANDRE L.R. CHAVES

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.


2011 ◽  
Vol 46 (4) ◽  
pp. 362-369 ◽  
Author(s):  
Marcos Cesar Gonçalves ◽  
Diogo Manzano Galdeano ◽  
Ivan de Godoy Maia ◽  
César Martins Chagas

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.


2020 ◽  
Vol 18 (4) ◽  
pp. e10SC05
Author(s):  
Ivana Stankovic ◽  
Ana Vucurovic ◽  
Katarina Zecevic ◽  
Branka Petrovic ◽  
Danijela Ristic ◽  
...  

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.


2003 ◽  
Vol 93 (12) ◽  
pp. 1485-1495 ◽  
Author(s):  
S. Chakraborty ◽  
P. K. Pandey ◽  
M. K. Banerjee ◽  
G. Kalloo ◽  
C. M. Fauquet

The biological and molecular properties of Tomato leaf curl Gujarat virus from Varanasi, India (ToLCGV-[Var]) were characterized. ToLCGV-[Var] could be transmitted by grafting and through whitefly transmission in a persistent manner. The full-length genome of DNA-A and DNA-B of ToLCGV-[Var] was cloned in pUC18. Sequence analysis revealed that DNA-A (AY190290) is 2,757 bp and DNA-B (AY190291) is 2,688 bp in length. ToLCGV-[Var] could infect and cause symptoms in tomato, pepper, Nicotiana benthamiana, and N. tabacum when partial tandem dimeric constructs of DNA-A and DNA-B were co-inoculated by particle bombardment. DNA-A alone also is infectious, but symptoms were milder and took longer to develop. ToLCGV-Var virus can be transmitted through sap inoculation from infected tomato plants to the above-mentioned hosts causing the same symptoms. Open reading frames (ORFs) in both DNA-A and DNA-B are organized similarly to those in other begomoviruses. DNA-A and DNA-B share a common region of 155 bp with only 60% sequence identity. DNA-B of ToLCGV-[Var] shares overall 80% identity with DNA-B of Tomato leaf curl New Delhi virus-Severe (ToLCNDV-Svr) and 75% with ToLCNDV-[Lucknow] (ToLCNDV-[Luc]). Comparison of DNA-A sequence with different begomoviruses indicates that ToLCGV-[Var] shares 84% identity with Tomato leaf curl Karnataka virus (ToLCKV) and 66% with ToLCNDV-Svr. ToLCGV-[Var] shares a maximum of 98% identity with another isolate of the same region (ToLCGV-[Mir]; AF449999) and 97% identity with one isolate from Gujarat (ToLCGV-[Vad]; AF413671). All three viruses belong to the same species that is distinct from all the other geminivirus species described so far in the genus Begomovirus of the family Geminiviridae. The name Tomato leaf curl Gujarat virus is proposed because the first sequence was taken from an isolate of Gujarat, India.


2004 ◽  
Vol 22 (4) ◽  
pp. 799-800 ◽  
Author(s):  
Alice K. Inoue-Nagata ◽  
Jesús Navas-Castillo ◽  
Paulo C.T. de Melo ◽  
Antônio C. de Ávila

A doença causada pelo complexo de vírus do tomato yellow leaf curl (TYLC) é muito séria em tomateiro, principalmente na América Central e Europa, e é causada por um complexo de begomovírus monopartidos. A doença torna-se predominante, mesmo em áreas com a presença de outros begomovírus. No Brasil, os problemas advindos da infecção por begomovírus uram entre os principais fatores de perdas e oneração de custos. A introdução do complexo TYLC representa uma grande ameaça para os produtores. Este estudo visou a realização de testes de detecção baseados em reação de polimerase em cadeia (PCR) e hibridização específicos em amostras suspeitas coletadas no estado de São Paulo. Um total de 46 amostras com sintomas lembrando aqueles causados pelo complexo TYLC foram coletados no município de Campinas. Todas as amostras foram negativas para detecção de Tomato yellow leaf curl virus e Tomato yellow leaf curl Sardinia virus, as duas espécies mais importantes do completo TYLC. Este resultado sugeriu que as duas espécies de vírus ainda não foram introduzidas no Brasil ou que ainda não apresentam uma larga distribuição.


2001 ◽  
Vol 91 (2) ◽  
pp. 188-196 ◽  
Author(s):  
Murad Ghanim ◽  
Shai Morin ◽  
Henryk Czosnek

Whiteflies (Bemisia tabaci, biotype B) were able to transmit Tomato yellow leaf curl virus (TYLCV) 8 h after they were caged with infected tomato plants. The spread of TYLCV during this latent period was followed in organs thought to be involved in the translocation of the virus in B. tabaci. After increasing acquisition access periods (AAPs) on infected tomato plants, the stylets, the head, the midgut, a hemolymph sample, and the salivary glands dissected from individual insects were subjected to polymerase chain reaction (PCR) without any treatment; the presence of TYLCV was assessed with virus-specific primers. TYLCV DNA was first detected in the head of B. tabaci after a 10-min AAP. The virus was present in the midgut after 40 min and was first detected in the hemolymph after 90 min. TYLCV was found in the salivary glands 5.5 h after it was first detected in the hemolymph. Subjecting the insect organs to immunocapture-PCR showed that the virus capsid protein was in the insect organs at the same time as the virus genome, suggesting that at least some TYLCV translocates as virions. Although females are more efficient as vectors than males, TYLCV was detected in the salivary glands of males and of females after approximately the same AAP.


2007 ◽  
Vol 81 (11) ◽  
pp. 5902-5907 ◽  
Author(s):  
Linmei Ge ◽  
Jiangtao Zhang ◽  
Xueping Zhou ◽  
Hongye Li

ABSTRACT Geminiviruses have circular single-stranded DNA genomes and are important pathogens in tropical and subtropical regions, but their population diversity and variability are poorly understood. Here, we have investigated variations accumulating in Tomato yellow leaf curl China virus (TYLCCNV), a geminivirus in the genus Begomovirus of the family Geminiviridae. The population variation was analyzed in a naturally infected tomato (Solanum lycopersicom) plant and in Nicotiana benthamiana and tomato plants experimentally infected with a swarm of TYLCCNV DNA clones to provide an identical sequence for initiation of infection. Our results demonstrate that the population of TYLCCNV in a naturally infected tomato plant was genetically heterogeneous and that rapid mutation occurred in the populations amplified from N. benthamiana and tomato plants that had been infected with cloned DNA. This feature of the population of TYLCCNV in these plants consisted of the consensus sequence and a pool of mutants that are not identical but are closely related to the consensus sequence, and it coincides with the quasispecies concept described for many RNA viruses. The mutation frequency was circa 10−4 in N. benthamiana and tomato at 60 days postinoculation, a value comparable to that reported for plant RNA viruses. The quasispecies-like nature of the TYLCCNV populations suggested that TYLCCNV is capable of rapid evolution and adaptation in response to changing agricultural practices.


2016 ◽  
Vol 15 (2) ◽  
pp. 132
Author(s):  
Melinda . ◽  
Tri Asmira Damayanti ◽  
Sri Hendrastuti Hidayat

Molecular identification of bean common mosaic virus associated with yellow mosaic disease on yard long bean. Bean common mosaic virus (BCMV) has been reported as one of the causal agents of yellow mosaic disease on yard long bean in West Java and Central Java. Infected plants showed mosaic, yellowing, and mixture of yellow mosaic. The research was conducted to identify the diversity of BCMV associated with yellow mosaic disease based on coat protein (CP) gene sequences. Symptomatic leaf samples were collected from yard long bean growing areas in several districts in West Java (Bogor, Cirebon, Subang, and Indramayu), and several districts in Central Java (Tegal, Klaten, Solo, Yogjakarta, Sleman, and Magelang). Molecular detection using RT-PCR method was carried out by using specific primer to BCMV which will amplify the CP gene. DNA fragment, + 860 bp in size, was successfully amplified from 8 out of 13 leaf samples, i.e samples from three villages in Bogor District (Cangkurawok, Bubulak, Bojong), and five samples from District of Cirebon, Subang, Solo, Sleman, and Tegal. Sequence analysis of those DNA fragment showed that 4 isolates (Bogor-Cangkurawok, Subang, Solo and Sleman) had the highest homology to BCMV-BlC from Taiwan, whereas 2 isolates (Cirebon and Tegal) had the highest homology to BCMVNL1 from England. Further, phyllogenetic analysis revealed that those of 4 isolates were closely related to BCMV-BlC from Taiwan based on nucleotide as well as amino acid sequences; while those other 2 isolates were closely related to BCMV-NL1 from England based on nucleotide sequences but closely related to BCMV-BlC Y from China based on amino acid sequences. Phyllogenetic analysis showed that those of 6 BCMV isolates separated in two different clusters; 4 isolates (Bogor- Cangkurawok, Subang, Solo, and Sleman) in cluster 1 together with BCMV-BlC from Taiwan, while other 2 isolates (Cirebon and Tegal) in cluster 2 together with BCMV-NL1.


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