Journal
PLANT METHODS
Volume 15, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s13007-019-0502-0
Keywords
NLR-type immune receptor; Pathogen avirulence effector; Cell death; Race-specific disease resistance; Barley; Wheat; Leaf protoplasts
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Funding
- Max-Planck Society
- German Research Foundation in the Collaborative Research Centre [SFB670]
- Cluster of Excellence on Plant Sciences [1028]
- Horizon 2020 Framework Programme [742263]
- Daimler and Benz Foundation
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Background Plant disease resistance to host-adapted pathogens is often mediated by host nucleotide-binding and leucine-rich repeat (NLR) receptors that detect matching pathogen avirulence effectors (AVR) inside plant cells. AVR-triggered NLR activation is typically associated with a rapid host cell death at sites of attempted infection and this response constitutes a widely used surrogate for NLR activation. However, it is challenging to assess this cell death in cereal hosts. Results Here we quantify cell death upon NLR-mediated recognition of fungal pathogen AVRs in mesophyll leaf protoplasts of barley and wheat. We provide measurements for the recognition of the fungal AVRs AvrSr50 and AVR(a1) by their respective cereal NLRs Sr50 and Mla1 upon overexpression of the AVR and NLR pairs in mesophyll protoplast of both, wheat and barley. Conclusions Our data demonstrate that the here described approach can be effectively used to detect and quantify death of wheat and barley cells induced by overexpression of NLR and AVR effectors or AVR effector candidate genes from diverse fungal pathogens within 24 h.
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