Multiplexed effector screening for recognition by endogenous resistance genes using positive defense reporters in wheat protoplasts
© 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation.
| Veröffentlicht in: | The New phytologist. - 1979. - 241(2024), 6 vom: 23. Feb., Seite 2621-2636 |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , , , , , , |
| Format: | Online-Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
2024
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| Zugriff auf das übergeordnete Werk: | The New phytologist |
| Schlagworte: | Journal Article avirulence defense inducible promoters effectors luciferase plant defense resistance gene wheat protoplast |
| Zusammenfassung: | © 2024 The Authors. New Phytologist © 2024 New Phytologist Foundation. Plant resistance (R) and pathogen avirulence (Avr) gene interactions play a vital role in pathogen resistance. Efficient molecular screening tools for crops lack far behind their model organism counterparts, yet they are essential to rapidly identify agriculturally important molecular interactions that trigger host resistance. Here, we have developed a novel wheat protoplast assay that enables efficient screening of Avr/R interactions at scale. Our assay allows access to the extensive gene pool of phenotypically described R genes because it does not require the overexpression of cloned R genes. It is suitable for multiplexed Avr screening, with interactions tested in pools of up to 50 Avr candidates. We identified Avr/R-induced defense genes to create a promoter-luciferase reporter. Then, we combined this with a dual-color ratiometric reporter system that normalizes read-outs accounting for experimental variability and Avr/R-induced cell death. Moreover, we introduced a self-replicative plasmid reducing the amount of plasmid used in the assay. Our assay increases the throughput of Avr candidate screening, accelerating the study of cellular defense signaling and resistance gene identification in wheat. We anticipate that our assay will significantly accelerate Avr identification for many wheat pathogens, leading to improved genome-guided pathogen surveillance and breeding of disease-resistant crops |
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| Beschreibung: | Date Completed 23.02.2024 Date Revised 23.02.2024 published: Print-Electronic RefSeq: AJ311031.1 Citation Status MEDLINE |
| ISSN: | 1469-8137 |
| DOI: | 10.1111/nph.19555 |