Detection and discrimination of four Aspergillus section Nigri species by PCR.
Species of Aspergillus section Nigri are not easily distinguished by traditional morphological techniques, and typically are identified by DNA sequencing methods. We developed four PCR primers to distinguish between Aspergillus niger, Aspergillus welwitschiae, Aspergillus carbonarius and Aspergillus tubingensis, based on species-conserved differences in the calmodulin gene sequence. PCR amplification from total DNA using these primers was species specific; no amplification occurred from nontarget species DNA for each primer pair. Species-specific PCR could distinguish between species in mixed DNA templates, indicating a utility in determining culture uniformity of isolated Aspergillus strains. In addition, with these primer sets, each species could be detected in soil following mixed-species inoculation with Aspergillus spores. This indicates that PCR with these species-specific primers may be useful in determining the distribution of Aspergillus species in environmental samples without the need for species identification from isolated strains, as well as detecting species that may be infrequently isolated by culture-based methods.
Foodborne Toxin Detection and Prevention Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Albany, CA, USA.
J D
JD
Palumbo
Foodborne Toxin Detection and Prevention Research Unit, U.S. Department of Agriculture, Agricultural Research Service, Albany, CA, USA.
T L
TL
O'Keeffe
D001230:Aspergillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001234:Aspergillus niger / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002147:Calmodulin / Q000235:genetics; D017931:DNA Primers; D005800:Genes, Fungal; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology; D013045:Species Specificity
D002147:Calmodulin; D017931:DNA Primers
Aspergillus carbonarius; Aspergillus niger; calmodulin; fumonisin; grapes; ochratoxin; soil; species identification