scholarly journals Identificação de isolados de Citrus tristeza virus (CTV) protetivos para Citrus sinensis (L.) Osbeck

2018 ◽  
Vol 44 (1) ◽  
pp. 17-22
Author(s):  
Ana Paula Gonçalves ◽  
Karina Silva dos Santos ◽  
Camila de Cassia Silva ◽  
Tanara Garcia de Novaes ◽  
Rúbia de Oliveira Molina
Keyword(s):  
Citrus Sinensis ◽  
Citrus Tristeza Virus ◽  
Rt Pcr ◽  
Tristeza Virus ◽  
Citrus Tristeza

RESUMO O Citrus tristeza virus (CTV) causa significativas perdas na produtividade de laranja doce [Citrus sinensis (L.) Osbeck] e seu controle tem sido realizado principalmente com a premunização. O trabalho teve como objetivo analisar a variabilidade de isolados fortes e fracos de CTV provenientes de plantas de citros inoculadas e mantidas em casa de vegetação e amostras de campo, coletadas em pomar comercial situado no município de Rolândia, PR. Para a determinação da variabilidade e diversidade genética dos isolados foi realizada avaliação dos sintomas e empregadas as técnicas de RT– PCR e RFLP, utilizando os oligonucleotídeos específicos HCP1/HCP2 e posterior sequenciamento dos fragmentos amplificados. Na avaliação de canelura, os isolados mantidos em casa de vegetação induziram sintomas leves, com exceção do isolado severo Capão Bonito. Os sintomas mais severos ocorreram em amostras situadas no campo. De acordo com as análises multivariadas os isolados de CTV tendem a se agrupar conforme a severidade dos sintomas e condições ambientais as quais foram expostas formando agrupamentos distintos entre amostras provenientes do campo e casa de vegetação. O dendrograma gerado a partir do sequenciamento dos isolados e as análises multivariadas revelaram que o isolado proveniente da amostra “Forte Arapongas” apresentou maior similaridade com o controle padrão forte proveniente de Capão Bonito. Os isolados identificados como fracos e provenientes das amostras Pêra IAC e Rolândia 5 apresentaram maior similaridade. Pode-se aferir que plantas hospedeiras mantidas em campo possuem maior variabilidade de isolados.

scholarly journals Comportamento de híbridos de citros em relação à infecção natural pelo Citrus tristeza virus e à presença de sintomas de descamamento eruptivo

2014 ◽  
Vol 36 (3) ◽  
pp. 735-741 ◽  
Author(s):  
Almir Santos Rodrigues ◽  
Cristiane de Jesus Barbosa ◽  
Walter dos Santos Soares Filho ◽  
Juliana Freitas-Astúa
Keyword(s):  
Citrus Tristeza Virus ◽  
Rt Pcr ◽  
Tristeza Virus ◽  
Citrus Tristeza

O Programa de Melhoramento Genético de Citros da Embrapa Mandioca e Fruticultura vem gerando híbridos para utilização como porta-enxertos, que necessitam ser avaliados em relação ao comportamento frente à infecção natural por isolados locais de Citrus tristeza virus (CTV) e à presença de sintomas de descamamento eruptivo (BahiaBarkScaling disease - BBS). Este trabalho apresenta resultados da avaliação do comportamento de 141 híbridos (sob a forma de pés-francos ou enxertados) estabelecidos na área experimental da Embrapa Mandioca e Fruticultura, no Recôncavo Sul da Bahia. Foram avaliadas a presença e a severidade de sintomas de caneluras e descamamento por meio de escala de notas. Para detectar a presença do CTV, foi utilizado o método sorológico de ELISA indireto e RT-PCR. Os híbridos avaliados foram classificados como imunes, tolerantes e intolerantes ao CTV. A maioria dos híbridos que apresentaram sintomas de BBS tem uma tangerineira como parental.

scholarly journals Prevalence of Severe Strains of Citrus Tristeza Virus in Florida Citrus Nurseries

HortScience ◽  
1993 ◽  
Vol 28 (7) ◽  
pp. 699-700 ◽  
Author(s):  
Charles A. Powell ◽  
Robert R. Pelosi
Keyword(s):  
Citrus Sinensis ◽  
Citrus Tristeza Virus ◽  
Severe Strain ◽  
Florida Citrus ◽  
Tristeza Virus ◽  
Citrus Tristeza

Sixty-eight percent of the `Pineapple', 52% of the `Navel', 46% of the `Valencia', 38% of the `Hamlin', and 0% of the `Ambersweet' orange [Citrus sinensis (L.) Osh.] trees in five Florida citrus nurseries were infected with severe strains of citrus tristeza virus (CTV), as demonstrated by reaction with a monoclinal antibody specific for severe strains of the virus. Severe strains of CTV infected 4%, 46%, 76%, 30%, and 48% of the trees at each of the five nurseries, respectively, indicating a considerable difference in severe strain prevalence among the nurseries. Thirty-five percent of the trees in the scion blocks (budwood source) of the nurseries also contained severe strains of CTV.

scholarly journals Seleção de Isolados do Citrus tristeza virus (CTV) Protetivos para Laranja 'Pêra' [Citrus sinensis (l.) Osbeck] no Paraná

2015 ◽  
Author(s):  
Karina Silva dos Santos ◽  
Ana Paula Gonçalves ◽  
Carlos Henrique da Silva Gonçalves ◽  
Tanara Garcia de Novaes ◽  
Rubia de Oliveira Molina
Keyword(s):  
Citrus Sinensis ◽  
Citrus Tristeza Virus ◽  
Tristeza Virus ◽  
Citrus Tristeza

Detection of severe isolates os citrus tristeza virus by bidirectional RT-PCR (BD-RT-PCR)

2000 ◽  
Vol 5 (1) ◽  
Author(s):  
M. L. P. N. Targon ◽  
M. A. Machado ◽  
A. A. Souza ◽  
G. W. Müller
Keyword(s):  
Citrus Tristeza Virus ◽  
Rt Pcr ◽  
Tristeza Virus ◽  
Citrus Tristeza

scholarly journals Quantitative detection of Citrus tristeza virus in plant tissues and single aphids by real-time RT-PCR

2007 ◽  
Vol 120 (2) ◽  
pp. 177-188 ◽  
Author(s):  
Edson Bertolini ◽  
Aranzazu Moreno ◽  
Nieves Capote ◽  
Antonio Olmos ◽  
Ana de Luis ◽  
...  
Keyword(s):  
Real Time ◽  
Citrus Tristeza Virus ◽  
Quantitative Detection ◽  
Plant Tissues ◽  
Rt Pcr ◽  
Tristeza Virus ◽  
Citrus Tristeza

scholarly journals Development and Application of a Multiplex Reverse-Transcription Polymerase Chain Reaction Assay for Screening a Global Collection of Citrus tristeza virus Isolates

2010 ◽  
Vol 100 (10) ◽  
pp. 1077-1088 ◽  
Author(s):  
Avijit Roy ◽  
G. Ananthakrishnan ◽  
John S. Hartung ◽  
R. H. Brlansky
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Citrus Tristeza Virus ◽  
Phylogenetic Analyses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Variable Regions ◽  
Polymerase Chain ◽  
Tristeza Virus ◽  
Citrus Tristeza

The emerging diversity of Citrus tristeza virus (CTV) genotypes has complicated detection and diagnostic measures and prompted the search for new differentiation methods. To simplify the identification and differentiation of CTV genotypes, a multiplex reverse-transcription polymerase chain reaction (RT-PCR) technique for the screening of CTV isolates was developed. Variable regions within the open reading frame (ORF)-1a of diverse CTV genotypes were identified to develop first a simplex (S) and then a hexaplex (H) RT-PCR. CTV isolates have been grouped previously into five genotypes (namely, T3, T30, T36, VT, and B165) based on the nucleotide sequence comparisons and phylogenetic analyses. Nucleotide sequences from GenBank were used to design species and genotype-specific primers (GSPs). The GSPs were initially used for reliable detection of all CTV genotypes using S-RT-PCR. Furthermore, detection of all five recognized CTV genotypes was established using the H-RT-PCR. Six amplicons, one generic to all CTV isolates and one for each of the five recognized genotypes, were identified on the basis of their size and were confirmed by sequence analysis. In all, 175 CTV isolates from 29 citrus-growing countries were successfully analyzed by S- and H-RT-PCR. Of these, 97 isolates contained T36 genotypes, 95 contained T3 genotypes, 76 contained T30 genotypes, 71 contained VT genotypes, and 24 contained B165 genotype isolates. In total, 126 isolates contained mixed infections of 2 to 5 of the known CTV genotypes. Two of the CTV isolates could not be assigned to a known genotype. H-RT-PCR provides a sensitive, specific, reliable, and rapid way to screen for CTV genotypes compared with other methods for CTV genotype detection. Efficient identification of CTV genotypes will facilitate a better understanding of CTV isolates, including the possible interaction of different genotypes in causing or preventing diseases. The methods described can also be used in virus-free citrus propagation programs and in the development of CTV-resistant cultivars.

scholarly journals Improved Efficiency for Quantitative and Qualitative Indexing for Citrus tristeza virus and Citrus psorosis virus

Plant Disease ◽  
2007 ◽  
Vol 91 (9) ◽  
pp. 1089-1095 ◽  
Author(s):  
C. Rosa ◽  
M. Polek ◽  
B. W. Falk ◽  
A. Rowhani
Keyword(s):  
Citrus Tristeza Virus ◽  
Rna Extraction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Minimal Amount ◽  
Rt Pcr ◽  
Preparation Methods ◽  
Certification Programs ◽  
Citrus Psorosis Virus ◽  
Tristeza Virus ◽  
Citrus Tristeza

Reverse transcription–polymerase chain reaction (RT-PCR) assays were developed for the detection of Citrus tristeza virus (CTV; genus Closterovirus) and Citrus psorosis virus (CPsV; genus Ophiovirus) in citrus trees. Real-time TaqMan RT-PCR was also developed for the detection of CTV. Three different sample preparation methods were compared. The total RNA extraction method by Qiagen was found to be more reliable than the other two methods consisting of crude plant sap extraction and total nucleic acid trapping on a silica bed. Of 287 samples tested for CTV, 210 samples tested positive by RT-PCR and 198 samples by enzyme-linked immunosorbent assay (ELISA). Furthermore, the results from monthly tests of a selected number of field-grown CTV-infected trees showed that RT-PCR detected the virus in 100% of the infected trees in winter and summer, whereas ELISA did not. The one-tube RT-PCR detection was developed for CPsV and was more sensitive than ELISA. Notably, three of 10 CPsV isolates were not detected by ELISA. As demonstrated here, our approach allows the efficacious detection of different viruses in citrus plants using a minimal amount of tissue during all seasons. The molecular methods described could be used in citrus certification programs and to test trees in nurseries and commercial orchards.

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