Abstract: Virus-free test-tube seedlings of a potato cultivar, Quanyun No.4, were used in this study to analyze and optimize the cultivation conditions for producing original cultivar seeds.A complete split-plot experimental design was applied for the optimization on substrate formulation, planting density and fertilization in the experiment.The results indicated that the 3 factors were highly significant in affecting the seed yield, marketable tuber production and economic indicators.The chlorophyll content, leaf area indices, tuber yield and relevant economic considerations of the seeds grown on Formula A₄ (substrate in a ratio of vermiculite:perlite:sand=1:1:2) were the highest among all tested media.The planting density B₁ with a row spacing of 0.05 m×0.10 m effectively increased the yields on seeds and marketable potatoes with an improved financial return.The Fertilization C₂ applying fertilizer twice weekly increased the seed production while maintained a high percentage of marketable potatoes harvested.An extremely significant interaction among the substrate formulation, planting density and fertilization frequency was observed.In the end, the conditions for an efficient production of potato cultivar seeds would include a culture substrate consisting of vermiculite and sand in 1:1 ratio, a planting spacing between rows of 0.05 m×0.10 m, and fertilizer application twice a week.