Alignment of the ribosomal 5.8S-ITS amplicon sequences of representative <i>Aspergilli</i> isolates.

<p>A part of the amplicon sequences (nucleotides 101 to 200) alignment is presented. Restriction sites are presented as bold-underlined, and variable nucleotides are highlighted. The extra <i>Hha</i>I restriction site present in the Ai-1 sequence is also shown.</p

5.8S-ITS RFLP DNA fragment sizes profiles.

<p>Individual isolates and reference strain isolates from grapes are clustered according to their RFLP profile deduced after sequencing of the 5.8S-ITS amplicon. Fragment sizes generated respect to the restriction endonuclease site position in the sequence, are given in bp.</p

Ribosomal 5.8S-ITS region restriction digestion patterns of <i>Aspergillus</i> grape isolates.

<p>Restriction digestion patterns (designated as <b>A</b>, <b>B</b> and <b>C</b>) of ribosomal 5.8S-ITS DNA amplicons from various <i>Aspergilli</i> grape isolates (presented as isolate designations), after digestion with the restriction endonucleases <b><i>Hha</i></b><b>I</b> and <b><i>Hinf</i></b><b>I</b>. 50 bp <b>(</b><b><i>l</i></b><b>)</b> and 100 bp <b>(L)</b> DNA ladders are also shown.</p

Neighbour-Joining phylogenetic tree based on divergences of ribosomal 5.8S-ITS sequences.

<p>The alignment of ribosomal 5.8S-ITS sequences from 10 isolates and 10 reference strains of <i>Aspergillus</i> spp. was performed using the CLUSTALΩ program. Nucleotide divergences were estimated according to the Jukes–Cantor model. Node numbers represent the frequency (proportion) with which a cluster appears in 1000 bootstrap runs.</p

Correlation of HPLC and ELISA values for OTA determination.

<p>Linear correlation and <i>R²</i> for the two methods assayed for OTA quantification (ELISA & HPLC).</p

Restriction digestion patterns of sequenced ribosomal 5.8S-ITS region amplicons.

<p>Restriction digestion patterns (designated as <b>A</b>, <b>B</b>, <b>C</b> and <b>D</b>) of five sequenced ribosomal 5.8S-ITS DNA amplicons from five different <i>Aspergilli</i> grape isolates (presented as isolate designations), after digestion with the restriction endonucleases <b><i>Hha</i></b><b>I</b>, <b><i>Hinf</i></b><b>I</b> and <b><i>Rsa</i></b><b>I</b>. Each isolate is a representative of the five different <i>Aspergillus</i> species characterized in this study. <b>L</b>: DNA ladder.</p

<i>Aspergillus</i> section <i>Nigri</i> strains distribution.

<p><i>A</i>. section <i>Nigri</i> distribution in total mycoflora (bars) and different species distribution within the <i>A</i>. section <i>Nigri</i> group (percentages inside bars) for each sampled prefecture. The percentages underneath prefectures' names refer to distribution and incidence of <i>A</i>. section <i>Nigri</i>.</p

Identification, origin and OTA production of <i>A</i>. section <i>Nigri</i> isolates.

<p>Species identification and origin of <i>A</i>. section <i>Nigri</i> spp. isolated from the present study and their ochratoxigenic potential assayed by HPLC and ELISA methods.</p><p>Limit of Quantification (LOQ) 2 ng OTA g⁻¹ CYA and Limit of Detection (LOD) 1 ng OTA g⁻¹ CYA.</p