native or sterile grapes artificially inoculated with different
amounts of A. carbonarius conidia. The assay can detect up to 103
conidia per g of grape. Overall, 88 out of 125 samples proved positive
for fungal DNA presence. These samples, both natural and
inoculated grapes, were subsequently tested for A. carbonarius DNA
presence, using the AcKS10L/R primers (Selma et al., 2008). An
amplicon of 161 bp was detected in samples that proved positive
for fungal DNA (Fig. 2). A total of 37 out of 88 samples showed
presence of A. carbonarius DNA, however, the amplicon intensity
signal visualized in the agarose gel was different among positive
samples, indicating a different representation of the fungus in each
sample. For verification of the end point PCR assays and in order to
test for putative assay inhibition, all samples that proved either
positive or negative with ITS1/4 primers were subjected to PCR
using the BT_HKF/R primers (Crespo-Sempere et al., 2013) which
amplify a 180 bp fragment of the b-tubulin gene (Fig. 2).
native or sterile grapes artificially inoculated with differentamounts of A. carbonarius conidia. The assay can detect up to 103conidia per g of grape. Overall, 88 out of 125 samples proved positivefor fungal DNA presence. These samples, both natural andinoculated grapes, were subsequently tested for A. carbonarius DNApresence, using the AcKS10L/R primers (Selma et al., 2008). Anamplicon of 161 bp was detected in samples that proved positivefor fungal DNA (Fig. 2). A total of 37 out of 88 samples showedpresence of A. carbonarius DNA, however, the amplicon intensitysignal visualized in the agarose gel was different among positivesamples, indicating a different representation of the fungus in eachsample. For verification of the end point PCR assays and in order totest for putative assay inhibition, all samples that proved eitherpositive or negative with ITS1/4 primers were subjected to PCRusing the BT_HKF/R primers (Crespo-Sempere et al., 2013) whichamplify a 180 bp fragment of the b-tubulin gene (Fig. 2).
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