http://www.cnr.it/ontology/cnr/individuo/prodotto/ID56755

PCR-based Detection of Spiroplasma Citri Associated with Citrus Stubborn Disease (Articolo in rivista)

Type

Articolo in rivista (Classe)
Prodotto della ricerca (Classe)

Label

PCR-based Detection of Spiroplasma Citri Associated with Citrus Stubborn Disease (Articolo in rivista) (literal)

Anno

2008-01-01T00:00:00+01:00 (literal)

Alternative label

Yokomi, R.K., Mello, A., Saponari, M., Fletcher, J. (2008)
PCR-based Detection of Spiroplasma Citri Associated with Citrus Stubborn Disease
in Plant disease
(literal)

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Yokomi, R.K., Mello, A., Saponari, M., Fletcher, J. (literal)

Pagina inizio

253 (literal)

Pagina fine

260 (literal)

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92 (literal)

Rivista

Plant disease (Rivista)

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IMPACT FACTOR 2007: 1.790 (I.F. 2008 non é ancora stato pubblicato dall'ISI WOK ) (literal)

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Improvements in polymerase chain reaction (PCR) detection of Spiroplasma citri, the causal agent of citrus stubborn disease, made PCR more reliable than culturing for S. citri detection. Primer sequences from the P89 putative adhesin gene, which is present on a plasmid as well as in the S. citri genome, and the P58 putative adhesin multigene were found to be much more sensitive than spiralin gene primers. PCR was compared to isolations and culturing for stubborn detection in two 20 A citrus plots in northeastern Kern Co. Of the 761 trees tested, PCR matched culture results 93.8% of the time. This validation now supports the use of PCR for screening S. citri infection with subsequent confirmation by culturing and dark field microscopy. Quantitative, real time PCR was also developed for S. citri detection using two different P58 primer sets. These primers identified two different S. citri strains in the citrus plots. Research is continuing to determine if these strains have a different symptom phenotype or vector transmissibility. (literal)

Note

ISI Web of Science (WOS) (literal)

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Yokomi R.K.: USDA, ARS, SJVASC, 9611 S. Riverbend Ave., Parlier, CA 93648 USA Mello,, A.F., Fletcher,, J.: Dept. Entomology and Plant Pathology, Oklahoma State University, Stillwater, OK 74078, U.S.A (literal)

Titolo

PCR-based Detection of Spiroplasma Citri Associated with Citrus Stubborn Disease (literal)

Abstract

Polymerase chain reaction (PCR)-based detection of citrus stubborn disease was improved using primers based on sequences of the P89 putative adhesin gene and the P58 putative adhesin multigene of Spiroplasma citri. Real-time PCR also was developed with detection limits estimated to be between 104 and 104 ng by serial dilution of a recombinant S. citri plasmid into DNA extracts from healthy Madam Vinous sweet orange. PCR for the detection of S. citri by these new primers was validated by comparing culturing of the pathogen, the traditional method of diagnosis, with PCR assays from samples taken from two citrus plots in Kern County, CA. Fruit columella was collected from 384 and 377 individual trees in each of two fields, respectively; one portion was used for culturing and the other for DNA extraction and PCR. PCR results matched those of culturing 85 to 100% of the time depending on the primers used. More importantly, PCR detected S. citri from culture-negative trees in 5 to 15% of the cases, suggesting that PCR performed as well or better than culturing for detection of S. citri in field samples. Real-time PCR proved to be the best method for detection. Differential reaction of the samples to the P58 primer pairs suggested that two populations of S. citri occur in historical and present-day field isolates. Citrus stubborn disease incidence was estimated to be 58.3 and 3.7% in the two orchards. The results presented here support the use of PCR for reliable detection of S. citri in field trees. (literal)

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