Author(s):
CECILE, FRANÇOIS 
; CASTAGNONE, CHANTAL ; BOONHAM, NEIL ; TOMLINSON, JENNY ; LAWSON, REBECCA ; HOCKLAND, SUE ; QUILL, JAMES ; VIEIRA, Paulo ; MOTA, Manuel ; CASTAGNONE-SERENO, PHILIPPE
Date: 2007
Persistent ID: http://hdl.handle.net/10174/6790
Origin: Repositório Científico da Universidade de Évora
Subject(s): Bursaphelenchus xylophilus; pinewood; nematode
Description
The pinewood nematode (PWN),
Bursaphelenchus xylophilus
, is
a major pathogen of conifers, which impacts on forest health,
natural ecosystem stability and international trade. As a
consequence, it has been listed as a quarantine organism in
Europe. A real-time PCR approach based on TaqMan chemistry
was developed to detect this organism. Specific probe and
primers were designed based on the sequence of the
Msp
I
satellite DNA family previously characterized in the genome of
the nematode. The method proved to be specific in tests with
target DNA from PWN isolates from worldwide origin. From a
practical point of view, detection limit was 1 pg of target DNA
or one individual nematode. In addition, PWN genomic DNA or
single individuals were positively detected in mixed samples in
which
B. xylophilius
was associated with the closely related
non-pathogenic species
B. mucronatus
, up to the limit of 0.01%
or 1% of the mixture, respectively. The real-time PCR assay was
also used in conjunction with a simple DNA extraction method to
detect PWN directly in artificially infested wood samples. These
results demonstrate the potential of this assay to provide rapid,
accurate and sensitive molecular identification of the PWN in
relation to pest risk assessment in the field and quarantine
regulation.
