The variation in length of the intergenic spacer (IGS) region of the ribosomal DNA repeat unit was examined in 63 accessions of wild barley, Hordeum spontaneum, and seven accessions of cultivated barley, Hordeum vulgare. The accessions of wild barley were collected from ecologically diverse climatic and edaphic microsites in Israel, and the barley cultivars were those grown in India. Sixteen spacer-length variants (slvs) observed in the present study presumably belonged to two known rDNA loci ( Rrn1 and Rrn2). Each accession had one or more variants, which together represented the rDNA phenotype. The rDNA phenotypes of wild barley accessions were widely diverse and differed substantially from those of cultivated barley. The slv phenotypes and the corresponding alleles were shown to be largely correlated with different climatic, edaphic and ecogeographical microsites and niches (the "Evolution Canyon" at Lower Nahal Oren, Mount Carmel; and Tabigha, Eastern Upper Galilee Mountains), so that a particular rDNA phenotype of an accession could be used to predict the climate and soil to which the accession belonged. This sharp microsite ecogeographic variation in ribosomal DNA appears adaptive in nature, and is presumably driven by climatic and edaphic natural selection.