Abstract: Cucumis sativus var. hardwickii Royle is the only extant wild subspecies of domesticated cucumber（Cucumis sativus var. sativus L.）. The wild cucumber contains several favorable traits including root-knot nematodes and CMV resistance，etc. It has a strong branching capability. The fruit shape of wild cucumber is oval and it tastes very bitter. In this study，we created a set of substitution lines using wild cucumber accession lines PI183967 as the donor and domesticated cucumber inbred Xintaimici as the receptor. During the process of backcrossing and self crossing，the wild cucumber genomic fragments were tracked using 62 SSR markers distributed in the cucumber genome. Subsequently，21 single-segment substitution lines and 10 double-segment substitution lines were obtained，which in total cover 96.81% of the wild cucumber genome. These substitution lines provided a new resource for the utilization of genes from wild cucumber and also the materials basis for the precise genetic analysis of quantitative traits in cucumber，as well as for the investigation of molecular mechanism of evolution from wild cucumber to domesticated cucumber.

Key words: wild cucumber, substitution lines, SSR markers, new germplasm resources