2 research outputs found
A PCR-based ‘molecular tool box’ for in planta differential detection of Verticillium dahliae vegetative compatibility groups infecting artichoke
12 pagesA multiplex-nested-PCR procedure was developed for
in planta
detection of
Verticillium dahliae
isolates infecting
artichoke and assessment of their vegetative compatibility groups (VCGs). PCR markers were identified and assigned to
V. dahliae
VCGs, including: i) a 334 bp marker amplified from VCG1A or VCG2B
334
isolates; ii) a 688 bp marker
amplified from VCG2A or VCG4B isolates; and iii) a 688 bp and a 964 bp PCR marker amplified from VCG2B
824
isolates. The infecting
V. dahliae
VCGs were identified in artichoke tissues according to specific patterns of amplified
markers after two rounds of PCR. The PCR-based ‘molecular tool box’ was first optimized using DNA extracted from
artichoke plants artificially inoculated with isolates representative of known VCGs. Thereafter, the efficiency of the
molecular procedure was tested using DNA extracted from naturally-infected artichoke plants showing a range of
symptom severity as well as from symptomless plants. The novel multiplex-nested-PCR assay was clearly superior in
detecting the pathogen compared to conventional isolation procedures, and in addition was informative about the VCGs.
Moreover, the PCR method allowed the detection and VCG identification of
V. dahliae
infections in symptomless but
infected plants, which had yielded false negatives when checked by microbiological isolation procedures. This ‘molecular tool box’ has uncovered the presence of several
V. dahliae
VCGs infecting the same artichoke plants in the Comunidad Valenciana Region. In addition, it is useful for genetic and pathogenicity diversity studies of V. dahliae populations infecting artichoke, and may help in predicting the severity of verticillium wilt epidemics.Comisión Interministerial de
Ciencia y TecnologÃa (CICYT) and ConsellerÃa de Agricultura, Pesca y
Alimentación de Generalitat Valenciana.Peer reviewe