Two cDNA clones encoding chitinase were isolated from a root nodule cDNA library of Elaeagnus umbellata by the hybridization-competition method. The two clones, EuNOD-CHT1 and EuNOD-CHT2, encode for 335 and 317 amino acid residues with the molecular mass of mature proteins being 33.3 and 31.1 kDa, respectively. The two chitinases showed similar protein structures consisting of four domains: hydrophobic signal peptide domain, cysteine-rich chitin-binding domain, hinge domain, and catalytic domain. The EuNOD-CHT1 gene showed similar expression levels in root nodules and leaves, with no detection of transcripts in the roots. The EuNOD-CHT2 gene was expressed at similarly high levels in the roots and root nodules, but at a very low level in the leaves. In situ hybridization showed that EuNOD-CHT1 transcripts were strongly detected in the meristem zone, but weakly detected in the outer cortex layer of the root nodule and in the uninfected cells of the fixation zone. On the other hand, EuNOD-CHT2 transcripts were strongly detected in the infected cells of the fixation zone and central vascular system, but weakly detected in the senescence zone. Our results suggest that the two chitinases may play different biological roles in the root nodule. EuNOD-CHT2 may be involved in a defense response against internal symbionts, external pathogens, or both, while EuNOD-CHT1 may be involved in normal plant development as well as in a defensive role against external pathogens.