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| Status |
Public on Mar 01, 2010 |
| Title |
Oligonucleoteide microarray for the identification of potential mycotoxigenic fungi |
| Platform organisms |
Aspergillus; Aspergillus amstelodami; Aspergillus clavatus; Aspergillus niger; Penicillium; Fusarium; Fusarium oxysporum; Fusarium sporotrichioides; Fusarium acuminatum; Fusarium graminearum; Alternaria alternata; Penicillium expansum; Talaromyces islandicus; Fusarium avenaceum; Aspergillus carbonarius; Fusarium subglutinans; Aspergillus versicolor; Fusarium cerealis; Fusarium equiseti; Penicillium fellutanum; Penicillium corylophilum; Fusarium verticillioides |
| Sample organisms |
Aspergillus amstelodami; Aspergillus clavatus; Aspergillus niger; Aspergillus parasiticus; Claviceps purpurea; Fusarium sambucinum; Curvularia lunata; Fusarium oxysporum; Fusarium sporotrichioides; Fusarium acuminatum; Fusarium graminearum; Alternaria alternata; Penicillium expansum; Talaromyces funiculosus; Talaromyces islandicus; Fusarium sp.; Fusarium avenaceum; Penicillium italicum; Aspergillus carbonarius; Aspergillus multicolor; Fusarium subglutinans; Bipolaris sorokiniana; Aspergillus versicolor; Fusarium anthophilum; Fusarium cerealis; Fusarium decemcellulare; Penicillium viridicatum; Fusarium equiseti; Drechslera sp.; Penicillium fellutanum; Penicillium corylophilum; Fusarium globosum; Fusarium verticillioides; Talaromyces rugulosus; Fusarium solani; Aspergillus chevalieri; Stenocarpella maydis; Fusarium incarnatum; Pseudopithomyces chartarum |
| Experiment type |
Expression profiling by array
Other
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| Summary |
Mycotoxins are secondary metabolites which are produced by numerous fungi and pose a continuous challenge to the safety and quality of food commodities in South Africa. These toxins have toxicologically relevant effects on humans and animals that eat contaminated foods. In this study, a diagnostic DNA microarray was developed for the identification of the most common food-borne fungi, as well as the genes leading to toxin production. A total of 40 potentially mycotoxigenic fungi isolated from different food commodities, as well as the genes that are involved in the mycotoxin synthetic pathways, were analyzed. For fungal identification, oligonucleotide probes were designed by exploiting the sequence variations of the elongation factor 1-alpha (EF-1 alpha) coding regions and the internal transcribed spacer (ITS) regions of the rRNA gene cassette. For the detection of fungi able to produce mycotoxins, oligonucleotide probes directed towards genes leading to toxin production from different fungal strains were identified in data available in the public domain. The probes selected for fungal identification and the probes specific for toxin producing genes were spotted onto microarray slides. The diagnostic microarray developed can be used to identify single pure strains or cultures of potentially mycotoxigenic fungi as well as genes leading to toxin production in both laboratory samples and maize-derived foods offering an interesting potential for microbiological laboratories.
Keywords: Development of a diagnostic microarray for the identification of potentially mycotoxigenic fungi as well as genes leading to toxin production, 40 food-borne fungi, mycotoxins
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| Overall design |
Development of a diagnostic array for the identification of food-borne fungi and their potential mycotoxin-producing genes. Oligonucleotide probes to be printed onto the array were designed by exploiting the sequence variations of the elongation factor 1-alpha (EF-1 α) coding regions and the internal transcribed spacer (ITS) regions of the rRNA gene cassette. For the detection of fungi able to produce mycotoxins, oligonucleotides directed towards genes leading to toxin production from different fungal strains were identified in data available in the public domain. Analysis was performed with 40 fungal cultures were obtained from the Agricultural Research Council culture collection (ARC), Pretoria, South Africa.an in-house spotted oligonucleotide microarray. The identity of each fungus was confirmed by standard laboratory procedures. For DNA isolation, the fungal strains were grown on 1.5% malt extract agar at 25°C for 1-2 weeks and total genomic fungal DNA was extracted following the DNA extraction protocol described by Raeder and Broda (1985). The internal transcribed spacer oligonucleotides ITS1, ITS3 and ITS4 were used as a reference for normalization of all spot intensity data.Samples were fluorescently labelled with Cy5 dye by using a Cy™Dye Post-labelling Reactive Dye Pack and wre hybridized to the oligonucleotide microarray overnight. Two biological and one technical replicate (using independent labelling reactions) was performed, each replication consisting of a reverse labelling experiment.
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| Contributor(s) |
Lezar S, Barros E |
| Citation(s) |
20307326 |
| |
| Submission date |
Nov 30, 2009 |
| Last update date |
Mar 21, 2012 |
| Contact name |
Eugenia Barros |
| E-mail(s) |
slezar@csir.co.za
|
| Phone |
0027-12-8413221
|
| Fax |
0027-12-8413651
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| Organization name |
CSIR
|
| Department |
Building 20
|
| Lab |
Biosciences
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| Street address |
Meiring Naude Rd
|
| City |
Pretoria |
| State/province |
Gauteng |
| ZIP/Postal code |
0001 |
| Country |
South Africa |
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| Platforms (1) |
| GPL9730 |
CSIR Fungi 20090901 96 v1.0 |
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| Samples (120)
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| Relations |
| BioProject |
PRJNA120853 |
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