Plant Disease (2006) 90, 1523-1530

G. Licciardello, F.M. Grasso, P. Bella, G. Cirvilleri, V. Grimaldi and V. Catara (2006)
Identification and detection of Phoma tracheiphila, causal agent of citrus mal secco disease, by real-time polymerase chain reaction
Plant Disease 90 (12), 1523-1530
Abstract: Phoma tracheiphila is the causal agent of a tracheomycotic disease of citrus called mal secco causing the dieback of twigs and branches. This pathogen is of quarantine concern; therefore, fast and reliable protocols are required to detect it promptly. A specific primer pair and a dual-labeled fluorogenic probe were used in a real-time polymerase chain reaction (PCR) with the Cepheid Smart Cycler II System (Transportable Device TD configuration) to detect this fungus in citrus samples. Real-time PCR assay was compared to modified conventional PCR assay. The sensitivity of the former was evaluated by testing P. tracheiphila DNA dilutions, and the minimum amount detectable was about 500 fg, whereas the linear quantification range was within 100 ng to 1 pg. Conventional PCR sensitivity was 10 pg. Conventional and real-time PCR successfully detected the fungus in woody samples of naturally infected lemon and artificially inoculated sour orange seedlings. Nevertheless, real-time PCR was about 10- to 20-fold more sensitive than conventional PCR, and preliminary results indicate that the former technique achieves quantitative monitoring of the fungus in tissues. Simple and rapid procedures to obtain suitable DNA samples from fungal cultures and citrus woody samples for PCR assays enable diagnosis to be completed in a short time.
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Database assignments for author(s): Vittoria Catara, Patrizia Bella, Gabriella Cirvilleri, Virgilio Grimaldi

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
quarantine treatments/regulations/aspects

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Plenodomus tracheiphilus	Citrus (genus)