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Title: Nodule‐specific PLAT domain proteins are expanded in the Medicago lineage and required for nodulation

Symbiotic nitrogen fixation in legumes is mediated by an interplay of signaling processes between plant hosts and rhizobial symbionts. In legumes, several secreted protein families have undergone expansions and play key roles in nodulation. Thus, identifying lineage‐specific expansions (LSEs) of nodulation‐associated genes can be a strategy to discover candidate gene families.

Using bioinformatic tools, we identified 13LSEs of nodulation‐related secreted protein families, each unique to eitherGlycine,ArachisorMedicagolineages. In theMedicagolineage, nodule‐specific Polycystin‐1, Lipoxygenase, Alpha Toxin (PLAT) domain proteins (NPDs) expanded to five members. We examinedNPDfunction usingCRISPR/Cas9 multiplex genome editing to createMedicago truncatulaNPDknockout lines, targeting one to fiveNPDgenes.

Mutant lines with differing combinations ofNPDgene inactivations had progressively smaller nodules, earlier onset of nodule senescence, or ineffective nodules compared to the wild‐type control. Double‐ and triple‐knockout lines showed dissimilar nodulation phenotypes but coincided in upregulation of aDHHC‐type zinc finger and an aspartyl protease gene, possible candidates for the observed disturbance of proper nodule function.

By postulating that gene family expansions can be used to detect candidate genes, we identified a family of nodule‐specificPLATdomain proteins and confirmed that they play a role in successful nodule formation.

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Author(s) / Creator(s):
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Date Published:
Journal Name:
New Phytologist
Page Range / eLocation ID:
p. 1538-1550
Medium: X
Sponsoring Org:
National Science Foundation
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