Development of a specific TaqMan real-time PCR assay for quantification of Fusarium graminearum clade 7 and comparison of fungal biomass determined by PCR with deoxynivalenol content in wheat and barley.
A Fusarium graminearum clade 7 specific real-time quantitative PCR (qPCR) assay was developed in this study based on unique polymorphisms in sequences of the mating type protein (MAT) gene. PCR amplification was not observed in eight phylogenetic lineages of the F. graminearum complex and four other closely related Fusarium species. Accuracy of the quantification of the real-time PCR assay was verified with wheat DNA spiked with F. graminearum clade 7 DNA. Wheat samples representing two Canadian wheat classes, CWRS (Canadian Western Red Spring) and CWRW (Canadian Western Red Winter) were used to determine the relationships among F. graminearum DNA, deoxynivalenol (DON) and Fusarium damaged kernel (FDK). The amount of DON and F. graminearum DNA remaining after removal of FDK varied among samples, but was sometimes substantial. Positive correlations were observed between F. graminearum clade 7 DNA (in picograms) and DON as well as FDK. There was also a strong correlation between FDK and DON in CWRS and CWRW wheat composite samples, but the inherent variability in individual producer samples precluded a definitive correlation. For barley, a positive correlation was observed between Fusarium DNA and DON values. Real-time PCR assays can be a valuable tool for barley as there are no reliable symptoms to visually assess the level of Fusarium head blight in this crop.
Demeke
Tigst
T
Canadian Grain Commission, Winnipeg, Manitoba, Canada. tigst.demeke@grainscanada.gc.ca
Gräfenhan
Tom
T
Clear
Randy M
RM
Phan
Anh
A
Ratnayaka
Indira
I
Chapados
Julie
J
Patrick
Susan K
SK
Gaba
Don
D
Lévesque
C André
CA
Seifert
Keith A
KA
D001483:Base Sequence; D018533:Biomass; D004271:DNA, Fungal / Q000032:analysis*; D005656:Fungal Proteins / Q000235:genetics; D005670:Fusarium / Q000235:genetics*; D005838:Genotype; D001467:Hordeum / Q000382:microbiology*; D010802:Phylogeny; D010935:Plant Diseases / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D015203:Reproducibility of Results; D012639:Seeds / Q000382:microbiology; D017422:Sequence Analysis, DNA / Q000379:methods; D014255:Trichothecenes / Q000032:analysis*; D014908:Triticum / Q000382:microbiology*
D004271:DNA, Fungal; D005656:Fungal Proteins; D014255:Trichothecenes; C007262:deoxynivalenol