Refinement of PCR‐detection of Fusarium avenaceum and evidence from DNA marker studies for phenetic relatedness to Fusarium tricinctum

Abstract

Existing PCR-based assays for the detection of the cereal pathogen Fusarium avenaceum were found to cross-react with F. tricinctum. An investigation into the phenetic relationship between these two species using two different marker systems revealed a close relationship between them despite their being from separate taxonomic sections. Whilst RFLP differences in the ITS regions surrounding the nuclear 5·8 S rDNA were insufficient to discriminate between isolates of the two species, they were clearly differentiated by RAPD profiling. RAPD fragments from F. avenaceum isolates were screened for hybridization to F. tricinctum DNA on Southern blots. One of 12 selected RAPD fragments showed no hybridization to genomic DNA from F. tricinctum. This fragment was cloned and sequenced, and the sequence obtained was used to design PCR primers. The primers were found to be specific for F. avenaceum, with no cross-reactions obtained with F. tricinctum or any other wheat pathogen assayed. The primers were able to differentiate between the two species in infected plant material, in contrast to the earlier assays.

Fusarium spp. In wheat grain in the Czech Republic analysed by PCR method

“…Turner et al (1998) was used, F. culmorum and F. graminearum were detected according to Schilling et al (1996), while F. poae was detected using the method of Parry and Nicholson (1996). The grain samples of different varieties of wheat (total number 160 samples) came from 3 grain purchase centres in southern Moravia and were collected in the years [2003][2004][2005][2006].…”

Fungi of the Fusarium genus in the grains of conventional hybrids and transgenic Bt-hybrids of maize (Zea mays L.) in the Czech Republic

“…DNA purity was evaluated by comparing the 260/280 absorbance ratio. For DNA amplifi cation, selected pairs of primers were used: JIAf/r for F. avenaceum (Turner et al, 1998), Fg16F/R for F. graminearum , Fp82F/R for F. poae (Parry & Nicholson, 1996), SUB1/2 for F. subglutinans, and VER1/2 for F. verticillioides (Mulè et al, 2004). Sequences of the specifi c primers used are presented in Tab.…”

Microbial analysis of a composted product of marine animal resources and isolation of bacteria antagonistic to a plant pathogen from the compost

“…used in isolation of antagonistic bacterium was kindly provided by Dr. Shinoyama (Meisei University, Japan). To determine the fungal species, the 5.8S nuclear rDNA together with its flanking internal transcribed spacer (ITS) sequences were amplified by using the following primers: ITS5 (5 -GGAAGTAAAAGTCGTAA CAAGG-3 ) and ITS4 (5 -TCCTCCGCTTATTGATATG C-3 ) (Turner et al, 1998). The resultant 691-bp-long fragment was cloned and its sequence was determined as mentioned above.…”