An atypical NLR pair CNL1/CNL5 from wild emmer confers powdery mildew resistance in wheat (PRJCA026280)

Resistance to wheat powdery mildew, a damaging foliar fungal disease, is commonly mediated by an individual resistance protein, most of which encode nucleotide-binding leucine-rich repeat (NLR) proteins. Here, we report that the powdery mildew resistance of Pm26 locus, originated from wild emmer wheat, is determined by a genetically linked atypical NLR pair CNL1/CNL5. Map-based cloning and PacBio HiFi long-read sequencing reveal CNL1 encodes an atypical coiled-coil domain containing NLR protein (CNL) fused with a novel NCKX domain, while CNL5 encodes a canonical CNL protein. Mutagenesis and VIGS experiments indicate that mutation or knock down expression of either CNL1 or CNL5 can lead to loss of resistance. Transgenic plants with CNL1 alone or CNL1/CNL5 together show resistance, while CNL5 alone not. Pm26 is geographical distributed mainly in wild emmer wheat Southern populations. Our study highlights atypical paired NLRs coordinately regulates powdery mildew resistance in wheat and provides diversified resistance gene resources for modern wheat breeding.