TY - JOUR
T1 - Specific detection of Fusarium langsethiae and related species by DGGE and ARMS-PCR of a β-tubulin (tub1) gene fragment
AU - Mach, R. L.
AU - Kullnig-Gradinger, C. M.
AU - Farnleitner, A. H.
AU - Reischer, G.
AU - Adler, A.
AU - Kubicek, C. P.
N1 - Funding Information:
This study was supported by grant nos. 1132 and 1264 from the Austrian Ministry of Agriculture and Forestry to CPK which is gratefully acknowledged. The authors are grateful to Dr. Kerry O'Donnell for the supply of strains of F. robustum, F. sambucinum and F. tumidum.
PY - 2004/9/15
Y1 - 2004/9/15
N2 - Fusarium langsethiae was recently described as a new toxigenic Fusarium species, which morphologically resembles Fusarium poae, but exhibits a mycotoxin pattern related to Fusarium sporotrichioides. To develop tools for early and specific detection of F. langsethiae and distinguishing it from related species of section Sporotrichiella and Discolor (F. poae, F. sporotrichioides, Fusarium kyushuense, Fusarium robustum, Fusarium sambucinum and Fusarium tumidum) sequence variations in their β-tubulin-encoding (tub1) gene were employed to design two PCR-based methods, denaturing gradient gel electrophoresis (DGGE) and amplification refractory mutation system (ARMS)-PCR. DGGE reliably separated all these strains, even from mixtures and in the presence of DNA from their natural hosts Zea mais, Triticum aestivum and Avena sativa. In addition, a tetraprimer ARMS-PCR, which employs two primer pairs to amplify, respectively, two different fragments of tub1 in a single PCR reaction resulted in rapid differentiation between F. langsethiae, F. sporotrichioides and F. poae according to the number of amplicons (four, two and one, respectively). These two methods will thus be worthwhile tools in the specific detection of F. langsethiae in infected crops.
AB - Fusarium langsethiae was recently described as a new toxigenic Fusarium species, which morphologically resembles Fusarium poae, but exhibits a mycotoxin pattern related to Fusarium sporotrichioides. To develop tools for early and specific detection of F. langsethiae and distinguishing it from related species of section Sporotrichiella and Discolor (F. poae, F. sporotrichioides, Fusarium kyushuense, Fusarium robustum, Fusarium sambucinum and Fusarium tumidum) sequence variations in their β-tubulin-encoding (tub1) gene were employed to design two PCR-based methods, denaturing gradient gel electrophoresis (DGGE) and amplification refractory mutation system (ARMS)-PCR. DGGE reliably separated all these strains, even from mixtures and in the presence of DNA from their natural hosts Zea mais, Triticum aestivum and Avena sativa. In addition, a tetraprimer ARMS-PCR, which employs two primer pairs to amplify, respectively, two different fragments of tub1 in a single PCR reaction resulted in rapid differentiation between F. langsethiae, F. sporotrichioides and F. poae according to the number of amplicons (four, two and one, respectively). These two methods will thus be worthwhile tools in the specific detection of F. langsethiae in infected crops.
KW - Amplification refractory mutation system (ARMS)-PCR
KW - Denaturing gradient gel electrophoresis (DGGE)
KW - Fusarium langsethiae
KW - Fusarium section Sporotrichiella
KW - Nucleic Acid Denaturation
KW - Species Specificity
KW - DNA, Fungal/chemistry
KW - Electrophoresis, Polyacrylamide Gel/methods
KW - Food Contamination/analysis
KW - Polymerase Chain Reaction/methods
KW - Fusarium/classification
KW - Base Sequence
KW - Genes, Fungal
KW - Tubulin/chemistry
UR - http://www.scopus.com/inward/record.url?scp=4444383059&partnerID=8YFLogxK
U2 - 10.1016/j.ijfoodmicro.2003.12.011
DO - 10.1016/j.ijfoodmicro.2003.12.011
M3 - Journal article
C2 - 15337597
AN - SCOPUS:4444383059
SN - 0168-1605
VL - 95
SP - 333
EP - 339
JO - International Journal of Food Microbiology
JF - International Journal of Food Microbiology
IS - 3
ER -