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Istituto Agronomico Mediterraneo di Bari (IAMB) 2003

Comparison between real-time Scorpion PCR and traditional methods for the detection and quantification of Verticillium dahliae in soils and infected olive trees

Barham, Hazar

Titre : Comparison between real-time Scorpion PCR and traditional methods for the detection and quantification of Verticillium dahliae in soils and infected olive trees

Auteur : Barham, Hazar

Etablissement de soutenance : Istituto Agronomico Mediterraneo di Bari (IAMB) - Centre international de hautes études agronomiques méditerranéennes (CIHEAM)

Diplôme : Master of science : Integrated pest management of Mediterranean fruit crops (2003)

Résumé
A technique based on the real-time Scorpion polymerase chain reaction for the detection of Verticillium dahliae in soil and xylem of infected olive trees was developed. Two specific primer pairs (Ver2 -Ver3 and Vd7b-Vd10), previously designed by comparing sequences from the ribosomal DNA intergenic spacer of V. dahliae and related species, were used. A nested-PCR protocol was applied to detect the pathogen by using Ver2-Ver3 in the first step and Vd7B-Vd10 in the second step. Primers specifity and detection limit were assessed by using genomic DNA from V. dahliae isolates and other species of the same genera. Specific Scorpion-PCR amplification, using modified Vd7b primer (Vd7b-FAM), was successful in detecting V. dahliae from soil and infected olive wood. The reliability of the entire procedure was assessed using both artificially and naturally infested soils and woody materials. Compared to traditional methods the molecular approach proved to be rapid, sensitive, and enabled a large-scale analysis.

Mots-clés : PCR VERTICILLIUM DAHLIAE OLIVES

Présentation (IAMB Library)

Page publiée le 20 juin 2014, mise à jour le 5 décembre 2017