Anne Edwards, Allan Downie JIC
Examination of ash petioles in Ashwellthorpe Lower Wood in Norfolk on 2nd June 2013 revealed the presence of fungal fruiting bodies similar in appearance to those produced by Hymenoscyphus pseudoalbidus and Hymenoscyphus albidus (see Figure) the sexual forms of the pathogen known as Chalara fraxinea.
Two of the fruiting bodies were tested using DNA primers reported to be specific for these two fungi. The ITS region of 18S rDNA was amplified using universal primers: 5’-TCCGTAGGTGAACCTGCGG-3’ and 5’-TCCTCCGCTTATTGATATGC-3’ and Chalara specific primers 5’-AGCTGGGGAAACCTGACTG-3’ and 5’-ACACCGCAAGGACCCTATC-3’. DNA sequencing of the PCR-amplified products has confirmed that these fruiting bodies are produced by the ash pathogen H. pseudoalbidus.
We believe this may be the first identification of spore-producing infections structures of the ash pathogen H. pseudoalbidus.in the UK and suggests that a new wave of infection has begun in the UK. It also fits with the identification of two mating types of the fungus in Norfolk.
Picture by Andrew Davis (JIC).
Contributors: Benjamin May, Mary Albury, Julia Shearman, Luke Young and Tony Moore University of Sussex
Material: We used the ‘Chalara_fraxinea_ass_s1v1_ann_v1.1.protein.faa’ & ‘Chalara_fraxinea_ass_s1v1_ann_v1.1.gene.gff’ Kenninghall wood assembly data within the ash dieback crowdsource github respository here )
Analysis: NCBI ORF finder blastp search MitoProtII SWISS-MODEL
Interpretation: ORF identified
A blastp search of the amino acid code using the FASTA sequence of Trypanosomal alternative oxidase (TAO) as a template found significant alignment with an E value of 6e-62.
MitoProt II identified a putative mitochondrial targeting sequence.
SWISS-MODEL was used to model the putative C. fraxinea AOX protein using trypanosomal alternative oxidase as a template (PDB 3VV9), with a QMEAN Z-score of -5.119, which is consistent with monotopic membrane proteins.