TWF
Funded by TWF


Projects

Molecular Mycology and Mycoparasitism
T.tip - Understanding mycoparasitic attack on the single cell level

Necrotrophic mycoparasitism of the genus Trichoderma is used in biological plant protection to destroy and antagonise soil-born plant pathogenic fungi. Fungal biocontrol agents (fungal BCAs) offer considerable advantages over the extensive use of synthetic fungicides with respect to human, animal and plant health and can be applied in environmentally sustainable ways.
The ‘Trichoderma tip’ project uses a novel approach for characterising the initiating steps of the mycoparasitic mode-of-action on the single cell level. Live-cell imaging and automated tip tracking analysis of CRIB reporter dynamics allow the identification of individual hyphae chemoattracted to host hyphae. Microcolony chemotropism assays quantify the chemotropic potential of isolated compounds involved in pre-contact host sensing.

This study will provide new insights towards (1) chemoattraction: in which parasite-host interactions is directional growth of individual hyphae an essential feature of the mycoparasitic attack?, (2) host specificity: which signalling molecules allow Trichoderma to differentiate between plant pathogenic host fungi?, and (3) physical contact: which conditions require the formation of specialised infection structures as part of a conserved killing mechanism? The cross-comparison between four different Trichoderma species and four model plant pathogenic host fungi will cover a wide range of agriculturally highly relevant fungus-fungus interactions, fill existing knowledge gaps in our understanding of the mycoparasitic attack program and ultimately contribute to an increased market availability of fungal BCAs in the future.

 

CRIB reporter in T. atroviride. Montage of a time sequence showing hyphal avoidance between tips with the CRIB reporter recruited to the apices. The Spitzenkörper and apical crescents of activated GTPases are clearly visible (asterisk). Scale bar, 10µm.

CRIB reporter in T. atroviride. Montage of a time sequence showing hyphal avoidance between tips that have the CRIB reporter recruited to the Spitzenkörper and the apical tip crescent. Scale bar, 10 µm. Spatio-temporal changes in GTPase activity are automatically followed and quantified by TipTracker software (see next image).

 

Tip Tracker
TipTracker v.3 Software. Developed by us for Neurospora crassa (Lichius et al., JCS 2014), was adapted for Trichoderma in collaboration with Prof. Mark Fricker, University of Oxford, and extended to the most recent version 3 which is able to track and quantify intensity changes of CRIB reporters in multiple hyphae within the same time course in parallel.

 

Leading hyphae of T. atroviride respond with growth arrest upon first chemical contact to Botrytis. Montage of the hyphal interaction zone between T. atroviride (CRIB-GFP) and B. cinerea (cytoplasmic DsRed). The total width of the shown confrontation zone is 3mm. Only the two hyphae at the very right hand corner recruit CRIB to their tips (asterisks), and thus presumably have not yet come into chemical contact with the host. Scale bar, 500µm.
Leading hyphae of T. atroviride respond with growth arrest upon first chemical contact to Botrytis. Montage of the hyphal interaction zone between T. atroviride (CRIB-GFP) and B. cinerea (cytoplasmic DsRed). The total width of the shown confrontation zone is 3 mm. Only the two hyphae at the very right hand corner recruit CRIB to their tips (asterisks) because they have not yet come into chemical contact with the host. All other hyphae within the immediate contact zone show clear signs of polarity stress induced by the defense response of B. cinerea. Scale bar, 500µm.
Team

Alexander LichiusAlexander Lichius, PI

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Dubraska Coromoto Moreno RuizDubraska Coromoto Moreno Ruiz

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LauraLaura Hackl

International Collaborator
University of Oxford

Mark FrickerMark Fricker

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 Funded by TWF 2018-2020

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