Cyperus esculentus is a new economic crop for both oil and grain worldwide. However, no relevant studies was reported on C. esculentus accessions in China. In order to clarify the main biological related traits, we selected three typical tubers, ‘Yuyousha 3’ (mediumlong), ‘Yuyousha 2’ (mediumround) and ‘YYS4’ (large), of C. esculentus and treated with single plant. The root tip cells of C. esculentus in mitotic phase were treated by DAPI(4′,6diamidino2phenylindole) staining and fluorescence in situ hybridization with a telomere DNA probe to analyze chromosome numbers in the cells. The results showed that: (1) three yield relative traits, tiller, single tuber weight and number of tuber per plant were showed extremely significant difference among materials. Among them, the single tuber weight of ‘YYS4’ (1.98 g) was the 3.3 and 5.4 times compared with that of ‘Yuyousha 2’ (0.61 g) and ‘Yuyousha 3’ (0.37 g). The number of tubers per plant of ‘Yuyousha 3’ was 245, which were the 1.3 and 1.6 times that of ‘Yuyousha 2’ and ‘YYS4’ respectively. ‘Yuyousha 3’ has the highest tiller number, which were 2.1 times that of ‘Yuyousha 2’ and 1.5 times that of ‘YYS4’. (2) The quality indexes of the three materials were significantly different. ‘Yuyousha 2’ showed highest oil content (32.00%), the soluble sugar content of ‘Yuyousha 3’ (21.30%) was the highest, and the starch and protein content of ‘YYS4’ was the highest. (3) ‘Yuyousha 2’ showed the leaves net photosynthetic efficiency (18.17 μmol·m^-2·s^-1) was the highest, the transpiration rate was relatively lower, and the water use efficiency was the highest among the three materials. (4) The chromosome numbers of the 3 materials were the same, all 156, and the results of DAPI staining and telomere probe fluorescence in situ hybridization were completely consistent. The results can provide a reference for subsequent breeding utilization, karyotype analysis and genetic classification.