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Abstract
BackgroundPlant receptors with lysin motifs (LsyM) recognize microbial signals such as fungal chitin and lipo-chitooligosaccharidic Nod factors of nitrogen-fixing rhizobia. It is generally assumed that ligand-induced dimerization of LysM receptors is an essential step in activation of intracellular kinase domains and downstream signaling. Consequently, genes required for plant defense and establishment of symbiosis are expressed. We recently found that three LysM receptor proteins (namely LYK1, LYK4 and LYK5) of Arabidopsis thaliana form a tripartite receptor complex to perceive chitin. However, constitutive and ligand-induced interactions of LysM receptors generally remain difficult to be characterized.ResultsInteractions between ectodomains of LYK1, LYK4 and LYK5 were investigated by a chimeric receptor approach using hairy roots of the legume Lotus japonicus. Synthetic receptor pairs consisting of a LYK ectodomain and the intracellular domain of a L. japonicus Nod factor receptor (NFR1 and NFR5, respectively) were tested for their capacity to activate expression of the symbiotic NIN (nodule inception) gene. The results indicated constitutive (LYK4ED–LYK4ED, LYK4ED–LYK5ED) and chitin-induced interactions (LYK1ED–LYK1ED, LYK1ED–LYK5ED) of the examined ectodomains.ConclusionWe present a method to functionally analyze constitutive and ligand-induced interactions of LysM-type proteins.
Highlights
Plant receptors with lysin motifs (LsyM) recognize microbial signals such as fungal chitin and lipochitooligosaccharidic Nod factors of nitrogen-fixing rhizobia
The hybrid protein constructs consisting of an A. thaliana LYK ectodomain (LYK1ED, 693-bp fragment of LYK1; LYK4ED, 813-bp fragment of LYK4; LYK5ED, 831-bp fragment of LYK5) and an intracellular domain of a L. japonicus Nod factor receptor of (NFR1ID, 1203-bp fragment of NFR1; NFR5ID, 1086-bp fragment of NFR5) were obtained by overlap extension PCR
The chimeric receptor method can provide clues to constitutive homodimerization of Lysin motif (LysM)‐type proteins Hairy roots of L. japonicus expressing a given chimeric receptor pair and a NINp-GUS construct (NIN promoter fused to the GUS reporter gene) were obtain by A. rhizogenes mediated transformation
Summary
Plant receptors with lysin motifs (LsyM) recognize microbial signals such as fungal chitin and lipochitooligosaccharidic Nod factors of nitrogen-fixing rhizobia. Rhizobial lipo-chitooligosaccharides, commonly known as Nod factors, and chitinous mycorrhizal molecules (chitin, chitooligosaccharides and lipo-chitooligosaccharides) are key signals to induce plant responses required for nodulation and mycorrhization [5, 6]. LysM receptors of plants are implicated in perception of microbial signals containing N-acetylglucosamine such as fungal chitin (or chitooligosaccharides), Nod factors and bacterial peptidoglycan [8,9,10]. Ligand-induced activation of the Nod factor receptor pair mediates induction of Nod factor signaling that includes various downstream components such as symbiosis receptor kinase (SymRK), calcium and calmodulin dependent protein kinase (CCaMK), its phosphorylation target (named CYCLOPS) and the key transcription regulator (nodule inception protein) NIN required for expression of Nod factor-induced genes [5, 17]. NIN expression is strongly up-regulated by Nod factor signaling in L. japonicus roots [17, 18]
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