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- Title
Agrobacterium-mediated insertional mutagenesis of the ochratoxigenic fungus Aspergillus westerdijkiae.
- Authors
Mata, Marcia M.; Taniwaki, Marta H.; Iamanaka, Beatriz T.; Sartori, Daniele; Oliveira, André L. M.; Furlaneto, Márcia C.; Fungaro, Maria Helena P.
- Abstract
Aspergillus westerdijkiae is a potent ochratoxin A (OTA) producer that has been found in coffee beans. OTA is known to have nephrotoxic effects and carcinogenic potential in animal species. Here we report for the first time the Agrobacterium-mediated transformation for Aspergillus westerdijkiae and the generation of ochratoxin-defective mutants. Conidia were transformed to hygromycin B resistance using strain AGL-1 of Agrobacterium tumefaciens. The obtained transformation frequency was up to 47 transformants per 106 target conidia. Among 600 transformants, approximately 5% showed morphological variations. Eight transformants with consistently reduced OTA production were obtained. Two of these transformants did not produce OTA (detection limit: 0.1 µg/kg); the other six mutants produced lower amounts of OTA (1%–32%) compared with the wild-type strain. By using thermal asymmetric interlaced polymerase chain reaction, we successfully identified a putative flavin adenine dinucleotide monooxygenase gene.
- Subjects
ASPERGILLUS; OCHRATOXINS; NEPHROTOXICOLOGY; ANIMAL species; AGROBACTERIUM; MONOOXYGENASES
- Publication
Canadian Journal of Microbiology, 2007, Vol 53, Issue 1, p148
- ISSN
0008-4166
- Publication type
Article
- DOI
10.1139/W06-100