Identification of the centromere-specific histone H3 variant in Lotus japonicus

• Centromeres contain a specific histone H3 variant.
• A homolog of the centromere-specific histone H3 is identified in Lotus japonicus.
• We localize GFP-LjCENH3 recombinant protein at centromeres.
• The study serves a foundation for future investigations on model legume centromeres.

The centromere is a structurally and functionally specialized region present on every eukaryotic chromosome. Lotus japonicus is a model legume species for which there is very limited information on the centromere structure. Here we cloned and characterized the L. japonicus homolog of the centromere-specific histone H3 gene (LjCenH3) encoding a 159-amino acid protein. Using an Agrobacterium-based transformation system, LjCenH3 tagged with a green fluorescent protein was transferred into L. japonicus cells. The centromeric position of LjCENH3 protein was revealed on L. japonicus metaphase chromosomes by an immunofluorescence assay. The identification of LjCenH3 as a critical centromere landmark could pave the way for a better understanding of centromere structure in this model and other agriculturally important legume species.