Doritis pulcherrima has 19 pairs of metacentric or submetacentric chromosomes which could not be distinguished from each other by conventional cytological techniques. Several repetitive DNA sequences including one tandem repeat consisting of 7-bp repeat units, two (GA)n microsatellite sequences were isolated from genomic DNA library of Doritis pulcherrima. These repetitive DNA sequences together with two ribosomal RNA genes and Arabidopsis-type telomeric sequences were mapped to mitotic metaphase chromosomes using fluorescence in situ hybridization. Based on the variation in the spatial and quantitative distribution of these repetitive sequences, the karyotype of Doritis pulcherrima was established.