@ARTICLE{TreeBASE2Ref16852, author = {Ludwig Niessen and Holger Schmidt and Rudi F. Vogel}, title = {The use of tri5 gene sequences for PCR detection and taxonomy of trichothecene-producing species in the Fusarium section Sporotrichiella.}, year = {2004}, keywords = {}, doi = {10.1016/j.ijfoodmicro.2003.12.009}, url = {}, pmid = {}, journal = {International Journal of Food Microbiology}, volume = {95}, number = {3}, pages = {305--319}, abstract = {Purified DNA from representative isolates each of Fusarium poae, F. sporotrichioides, F. kyushuense and the newly described F. langsethae was used to amplify a 658 bp fragment from the trichodiene synthase (tri5) gene of these fungi using the gene specific primer pair Tox5-1/Tox5-2. Fragments obtained were isolated and sequenced. The nucleotide sequences were aligned to each other. Sequence analysis revealed high similarity between F. sporotrichioides and F. langsethae (98.7 %). Similarity between the latter species and F. poae was lower (90.9 %). Phylogenetic analysis of the aligned sequences with the tri5 sequence of Stachybotrys chartarum as outgroup revealed a clear separation between F. poae and the group of F. sporotrichioides and F. langsethae sp. nov. Within this subgroup, two species were supported by high boostrap values.Oligonucleotide primers were designed from the aligned sequences and used as reverse primers in combination with the gene specific forward primer Tox5-1. The newly designed reverse primers enabled amplification of a 400 bp fragment when purified DNA from F. poae and F. sporotrichioides were used as template, respectively. Two different reverse primers designed for F. langsethae sp. nov. both could not differentiate this species from F. sporotrichioides but from F. poae and F. kyushuense. All primer pairs were tested for cross reactivity with 25 fungal species and subspecies capable of producing trichothecenes with no unspecific amplification found. The PCR assays developed were applied in analysis of artificially and naturally infected cereal grain. The species were selectively detected by the respective primers. In naturally infected oats, F. langsethae sp. nov. was identified by the combination of two PCR assays.} }

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