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Appl Environ Microbiol. 2000 Jun;66(6):2673-7.

Detection of Pseudomonas savastanoi pv. savastanoi in olive plants by enrichment and PCR.

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  • 1Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias, 46113 Moncada, Valencia, Spain.

Abstract

The sequence of the gene iaaL of Pseudomonas savastanoi EW2009 was used to design primers for PCR amplification. The iaaL-derived primers directed the amplification of a 454-bp fragment from genomic DNA isolated from 70 strains of P. savastanoi, whereas genomic DNA from 93 non-P. savastanoi isolates did not yield this amplified product. A previous bacterial enrichment in the semiselective liquid medium PVF-1 improved the PCR sensitivity level, allowing detection of 10 to 100 CFU/ml of plant extract. P. savastanoi was detected by the developed enrichment-PCR method in knots from different varieties of inoculated and naturally infected olive trees. Moreover, P. savastanoi was detected in symptomless stem tissues from naturally infected olive plants. This enrichment-PCR method is more sensitive and less cumbersome than the conventional isolation methods for detection of P. savastanoi.

PMID:
10831456
PMCID:
PMC110599
[PubMed - indexed for MEDLINE]
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