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The metabolic precursor of vitamin A, β-carotene, is essential for human health. The gene(s) controlling β-carotene quantity (QβC) has been introgressed from Xishuangbanna gourd (XIS, possessing β-carotene; Cucumis sativus L. var. xishuangbannanesis Qi et Yuan; 2 n = 2 x = 14) into cultivated cucumber (no β-carotene; Cucumis sativus L.). To determine the inheritance of QβC in cucumber fruit endocarp, F1 progeny and a set of 124 F7 recombinant inbred lines (RILs) derived from the cultivated cucumber line CC3 and XIS line SWCC8 were evaluated for QβC during 2009 and 2010 in Nanjing, China. Segregation analysis revealed that endocarp QβC of greenhouse-grown fruit was controlled by a single recessive gene. Further, marker analysis indicated the gene controlling QβC was linked to seven SSR markers on linkage group 3, where their order was SSR20710- SSR19511- SSR15419- SSR07706- ore- SSR23231- SSR11633- SSR20270. These markers and the putative candidate gene were mapped to cucumber chromosome 3DS. An evaluation of 30 genetically diverse cucumber lines indicated that marker SSR07706 has utility in further genetic analyses of the QβC orange endocarp gene, designated ore. Moreover, the markers defined herein may have utility for marker-assisted selection directed towards the development of cucumber germplasm with high fruit β-carotene content. [ABSTRACT FROM AUTHOR]