Uncovering the repertoire of legume NLR immune receptors Rita Maravilha Marques1,2, Carmen Santos1, Carlota Vaz Patto1, Sophien Kamoun2, and Jiorgos Kourelis2 1Instituto de Tecnologia Química e Biológica António Xavier, NOVA University of Lisbon, Oeiras 2780-157, Portugal 2The Sainsbury Laboratory, University of East Anglia, Norwich Research Park, NR4 7UH, Norwich, UK 1. Cardoso, D. B. O. S. et al. (2013). Reconstructing the deep-branching relationships of the papilionoid legumes. SA Journal of Botany, 89, 58-75. 2. Mundt, C. C. (2014). Durable resistance: a key to sustainable management of pathogens and pests. Infection, Genetics and Evolution, 27, 446-455. 3. de Queiroz, L. P. et al. (2015). A multilocus phylogenetic analysis reveals the monophyly of a recircumscribed papilionoid legume tribe Diocleae with well-supported generic relationships. Molecular Phylogenetics and Evolution, 90, 1-19. 4. Kourelis J et al. (2021) RefPlantNLR is a comprehensive collection of experimentally validated plant disease resistance proteins from the NLR family. PLOS Biology 19(10): e3001124. https:// doi.org/10.1371/journal.pbio.3001124. BUSCO Assesment Results 7. Conclusions and Outlook The number and distribution of NLR classes across legume species is variable and mostly reflects legume phylogeny Most legume species tend to have more CC-NLRs than TIR-NLRs The number of CCG10-NLRs and CCR-NLRs appears less variable Whole genome duplications and genetic erosion could be altering NLR numbers The Fabaceae NLR repertoire will be an important tool for comparative resistance mapping studies and for R gene enriched Genome-Wide Association Studies What is the nature and diversity of Legume NLRs? 2. Question and Aim Perform a comparative NLRome study 3. The completeness of legume proteomes could be improved 4. We built NLR phylogenies for each species using RefPlantNLR4 5. The abundance and distribution of NLR classes in the Fabaceae is variable 6. Legumes display expansions and contractions in NLR numbers <5% Missing = Good Assembly Max. likelihood trees - Clustal Omega Alignment - FastTree algorithm Phaseolus vulgaris NLR Phylogeny Example: Tree Scale: 1 Proteome + NLR tracker Genome Quality Control NLR Phylogenomics Fabaceae, the legume family, is a large and diverse angiosperm crop family, with much ecological and economic importance.1,3 However, biotic stress continues to seriously constrain legume yield2 Resistance breeding is considered the most sustainable plant disease management method2 Nucleotide-binding Leucine Rich genes (NLRs) are intracellular receptors that detect pathogen effectors, triggering a powerful defensive mechanism known as Hypersensitive Response4 The NLR resistance genes and their respective classes remain largely unexplored in legumes CC NBARC LRR CC NBARC LRR LRR TIR NBARC LRR LRR RPW8 NBARC NLR number according to total number of proteins Number of Protein-coding Loci Total Number of NLRs MLA36-1 MLA30-1 MLA7 MLA13 MLA12 2 MLA9 MLA10//MLA13//MLA30-1//MLA34//MLA36-1//MLA37-1//MLA7//MLA9 MLA10 MLA37-1 MLA3 MLA23 MLA35-1 MLA23//MLA3//MLA35-1 2 MLA34 MLA39-1 1 MLA39-1 2 MLA6 1 MLA6 2 MLA27-2 MLA2//MLA27-1//MLA27-2 MLA27-1 MLA2 MLA19-1 1 MLA19-1 2 MLA12 1 MLA18-2 2 MLA18-2 1 MLA31-1 1 MLA31-1 2 MLA32 1 MLA32 2 MLA22 1 MLA22 2 MLA8 MLA1//MLA8 MLA1 MLA28-1 1 MLA28-1 2 MLA25-1 1 2 MLA25-1 2 2 MLA16-1 1 2 MLA16-1 2 2 MLA18-1 1 2 MLA18-1 2 2 HcMLA1 1 HcMLA1 2 TmMLA1 1 TmMLA1 2 Sr33 1 2 Sr33 2 2 SrTA1662 2 SrTA1662 1 Sr50 2 Sr50 1 Pi36 1 Pi36 2 DES1 2 3 DES1 1 1 Sr35 2 2 Sr35 1 2 Yr-10 1 2 Yr-10 2 2 Pm1a 1 Pm1a 2 Sr22 2 2 Sr22 1 2 SrTm5 Sr22-PI190945//SrTm5 Sr22-PI190945 sr22-PI573523 2 sr22-PI573523 1 Lr10 1 Lr10 2 Sr13a Sr13a//Sr13b//Sr13c//Sr13d Sr13c Sr13d Sr13b Sr26 1 2 Sr26 2 2 NLS1 1 2 NLS1 2 2 Pbr1.b 1 Pbr1.b 2 Pbr1.c 1 2 Pbr1.c 2 2 Pm41 2 Pm41 1 Pm5b 2 4 Pm5b 1 1 Pm5e Pm5-BMDM Pm5-CS Pm5a Pm5-ND Pm5-BMDM//Pm5-CS//Pm5-ND//Pm5a//Pm5e RPG5 2 RPG5 1 YrU1 2 4 YrU1 1 1 Adnr1 2 Adnr1 1 Pi-ta 1 Pi-ta 2 RGA2a 1 RGA2a 2 Pia-2 1 1 Pi