Members of the Fusarium graminearum species complex are important cereal pathogens worldwide and belong to one of at least nine phylogenetically distinct species. We examined 298 strains of the F. graminearum species complex collected from wheat or barley in Japan to determine the species and trichothecene chemotype. Phylogenetic analyses and species-diagnostic polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLPs) revealed the presence and differential distribution of F. graminearum sensu stricto (s. str.) and F. asiaticum in Japan. F. graminearum s. str. is predominant in the north, especially in the Hokkaido area, while F. asiaticum is predominant in southern regions. In the Tohoku area, these species co-occurred. Trichothecene chemotyping of all strains by multiplex PCR revealed significantly different chemotype compositions of these species. All 50 strains of F. graminearum s. str. were of a 15- or 3-acetyl deoxynivalenol type, while 173 (70%) out of 246 strains of F. asiaticum were of a nivalenol type. The possibility of gene flow between the two species was investigated by use of 15 PCR-RFLP markers developed in this study. However, no obvious hybrids were detected from 98 strains examined, including strains collected from regions where both species co-occur.

The Fusarium graminearum species complex (Fg complex) that consists of at least 11 phylogenetically distinct species contains important Fusarium head blight (FHB) pathogens of wheat and barley worldwide. We obtained members of the Fg complex by sampling from a 500-m2 experimental wheat field in Kumamoto Prefecture, Japan in two consecutive years and assessed them for species identity and trichothecene chemotype. Haplotype diversity was estimated by using 11 variable numbers of tandem repeat (VNTR) markers. In addition to these two samples (group 03W in 2003 and group 04W in 2004), pathogen populations from seed that was harvested in Fukuoka Prefecture and planted in the experimental field in 2002 (group 02WSC) and pathogen populations from seed that was harvested in Nagasaki Prefecture and planted in 2003 (group 03WSC) were analyzed for this study. Forty-six isolates were collected in each group. Most isolates from wheat heads were classified as F. asiaticum; only four isolates were classified as F. graminearum sensu stricto (s. str.). Out of a total of 183 Fg complex strains, 80 isolates (44%) were of the NIV type, while 103 isolates (56%), including all four F. graminearum s. str. isolates, were of the 3ADON type. No 15ADON type isolate was detected in this study. Trichothecene chemotype compositions of 03W and 04W were nearly identical. High gene diversity of F. asiaticum was observed in all groups. Based on the observed low level of fixation index (FST) and high level of effective number of migrants (Nm), distinctive population subdivision of F. asiaticum was not inferred among the four groups. These results suggest that FHB in the experimental wheat field in both 2003 and 2004 was caused by a genetically similar population, which prevails in Kumamoto, Fukuoka, and Nagasaki prefectures.

Genetic subdivision of Fusarium asiaticum was investigated using a collection of 478 isolates originating from the Kyushu area and Aichi Prefecture, Japan and Zhejiang Province in China. Trichothecene-type determination by a multiplex PCR-test indicated that all isolates were either of a nivalenol (NIV) or a 3-acetyl deoxynivalenol (3ADON) type. The 15-acetyl deoxynivalenol (15ADON) type was not detected in this collection. Based on a Bayesian model-based clustering method using allele data obtained with 11 variable number of tandem repeats (VNTR) markers, we detected three genetic clusters. The majority of isolates in the clusters were NIV isolates from both Japan and China, Japanese 3ADON and Chinese 3ADON isolates, respectively. High levels of fixation indices and low levels of effective number of migrants were observed between the genetic clusters. Data was reanalyzed by classifying the isolates into six groups according to trichothecene type and geographic location. Population analyses of these re-classified groups indicated that the genetic subdivisions of F. asiaticum were correlated with both trichothecene type and geographic differences.

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