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High-resolution cytogenetic map can provide not only important biological information on genome organization but also solid foundation for genetic and genomic research.The progress in the molecular and cytogenetic studies has created the basis for developing the cytogenetic map in cucumber (Cucumis sativus L.).Here, the cytogenetic maps of four cucumber chromosomes (chromosomes 1, 3-5) were constructed by fluorescence in situ hybridization (FISH) analysis on cucumber pachytene chromosomes.