PMID:21077124 Localisation of the benzimidazole fungicide binding site of Gibberella zeae β2-tubulin studied by site-directed mutagenesis.

MPiovesana commented 1 year ago

Curated by @MPiovesana https://canto.phi-base.org/curs/3e64d02478d528b4

MPiovesana commented 1 year ago

In this paper, authors generate mutants of Gibberella zeae (Fusarium graminearum) carrying mutant alleles of the β2-tubulin gene. The mutant alleles are generated by site-directed in vitro mutagenesis of the WT β2-tubulin allele from strain 2021, and are introduced in strain DN83, which is strain 2021 harbouring the deletion of β2-tubulin. All mutant lines display increased resistance to benzimidazole fungicides and not fitness penalties. Interestingly, the deletion of β2-tubulin (strain DN83) led to a reduction in virulence.

MPiovesana commented 1 year ago

Uniprot ID: a gene and species name search retrieved ID Q4HZS8.

Strain: strain 2021 was selected for this curation session, as it is the recipient strain for transformation experiments following the deletion of the endogenous wild type β2-tubulin gene. Authors name this strain DN83, but the only difference between this and 2021 is the deletion of β2-tubulin. So here I suggest using 2021 as recipient as recording β2tubΔ as background.

Host: wheat cv. Yangmai 49 was added to the curation session.

MPiovesana commented 1 year ago

Genotype creation: as the mutant alleles were generated by in vitro mutagenesis of the wt β2-tubulin of strain 2021 (which is also the recipient strain), amino acid substitution alleles were created to record these genotypes. The deletion of the endogenous β2-tubulin was recorded as β2tubΔ in background. The mutant genotype β2tubΔ was also created as authors describe the phenotypes of this line. As authors show that expression of the mutant alleles is similar to that of the WT in the parental strain (RT-PCR in Fig 2), I selected 'wild type product level' under expression levels.

Genotype annotations: the following PHIPO terms were used resistance to carbendazim (AE low, medium and high used according to the text) sensitive to carbendazim

Authors also test the resistance of mutant lines to other benzimidazole fungicides (benomyl, thiabendazole and thiophanate-methyl), with similar results. I was not sure if it was necessary to add all these annotations, as all mutant genotypes are resistant to all fungicides and I thought it could become repetitive. So only carbendazim phenotypes recorded so far.

AC: comment added 'Similar results were obtained with the other benzimidazoles, benomyl, thiabendazole and thiophanate-methyl'

MPiovesana commented 1 year ago

Metagenotype creation: metagenotypes were created including the WT (2021), β2tubΔ and β2tub mutant genotypes + wheat were created.

Metagenotype annotation: the control wild type and mutant metagenotypes (except for β2tubΔ) were annotated with term "presence of pathogen-associated host lesions", as mutant lines displayed unaffected pathogenicity (recorded as annotation extension). The metagenotype composed of the β2tubΔ and wheat plants was annotated with "decreased extent of pathogen-associated host lesions", as this line displayed significantly reduced virulence (also recorded as AE).

MPiovesana commented 1 year ago

Curation completed pending review.