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(Referência obtida automaticamente do Google Scholar, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

In situ probing of Xylella fastidiosa in honeydew of a xylem sap-feeding insect using 16S rRNA-targeted fluorescent oligonucleotides

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Autor(es):
Rodrigues‚ J.L.M. ; Silva-Stenico‚ M.E. ; De Souza‚ A.N. ; Lopes‚ J.R.S. ; Tsai‚ S.M.
Número total de Autores: 5
Tipo de documento: Artigo Científico
Fonte: ENVIRONMENTAL MICROBIOLOGY; v. 8, n. 4, p. 747-754, 2006.
Resumo

Xylella fastidiosa is a plant pathogen that threatens a US$ 4.6 billion worldwide wine and citrus industry. Monitoring its presence and distribution in plants and vectors is crucial for designing control strategies, as well as for understanding its ecological role and fate. We developed two fluorescent oligonucleotide probes complementary to different regions of the 16S rRNA gene of X. fastidiosa. The specificity of the newly designed probes S-S-X.fas-0067-a-A-18 and S-S-X.fas-1439-a-A-18 was demonstrated using fluorescence in situ hybridization (FISH) for 12 Xylella isolates, 15 closely related microorganisms and three plant endophytes. These probes were used to detect and quantify X. fastidiosa in plant sap (average value of 2.9 +/- 0.3 x 10(6) cells ml(-1)) from three different citrus orchards. In a second experiment, cells were quantified in honeydew (2.2 +/- 0.2 x 10(4) cells ml(-1)) collected from the insect vector Bucephalogonia xanthophis during the acquisition access period on an infected plant. The number of pathogen cells retained or digested by the insect is 10 000 times greater than the estimated minimum value to ensure an efficient transmission. Polymerase chain reaction (PCR) amplification using specific primers with plant sap and honeydew samples, followed by sequencing, confirmed the presence of the plant pathogen. This is the first demonstration of FISH being used for environmental samples, such as plant sap and insect honeydew, to estimate the abundance of a plant pathogen during infection. (AU)

Processo FAPESP: 98/16265-1 - Functional analysis of Xylella genome by identification of proteins and putative virulence related low molecular weight compounds
Beneficiário:Tsai Siu Mui
Modalidade de apoio: Auxílio à Pesquisa - Programa GENOMA
Processo FAPESP: 00/10168-6 - Agronomical & Environmental Genomes
Beneficiário:Tsai Siu Mui
Modalidade de apoio: Auxílio à Pesquisa - Programa GENOMA
Processo FAPESP: 00/13160-6 - Detecção e qualificação de Xylella fastidiosa com o uso de real time (PCR) e hibridização fluorescente in situ (FISH) usando os genes que codificam para o 16S rRNA e virulência
Beneficiário:Jorge Luiz Mazza Rodrigues
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
Processo FAPESP: 00/08045-3 - Investigação e caracterização in vitro de toxinas/oligopeptídeos não-ribossomais do sobrenadante de Xylella fastidiosa
Beneficiário:Maria Estela Stenico
Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado