The production of flowers has increased in Brazil and has gained more economic importance, especially the chrysanthemum (Dendranthema grandiflora), which responds for approximately 80 % of the total internal flower market. The substrate is a basic input for the production of seedlings that results in plants of good quality, especially in vegetative propagated crops. A study was carried out under greenhouse conditions in randomized block design in 4x2 factorial scheme with the objective to test 4 substrates (S1 - commercial substrate TOPSTRATO HORTALIÇAS®; S2 - 50 % of commercial substrate mixed with 50 % of clay loam soil, S3 - 55 % of clay loam soil mixed with 15 % of washed sand, 15 % of cattle manure and 15 % of wood shavings, and S4 - 25 % of clay loam soil mixed with 25 % of washed sand, 25 % of cattle manure and 25 % of wood shavings, all on volume basis) in combination with pretreatment of stakes with STIMULATE®, growth promoter, in order to produce chrysanthemum cultivar Double Sorted seedlings. For analysis of the growth variables, chrysanthemum stakes with 5 leaves were planted individually in poly tubes. The poly tubes were covered with black screen of 50% shade and irrigated four times a day. At the end of experimental period the seedlings were harvested and growth variables were measured. Analysis of variance revealed significant effect (p ≤ 0.05) of the substrates in plant height (PH) and root length (RL). Substrate S2 presented best results, the PH and RL under this treatment being 12.10, 31.45 and 30.24 %, and 2.75, 18.81 and 14.16 % greater than S1, S3 and S4 substrates, respectively. On the other hand, the stakes without immersion in STIMULATE® showed better growth and significant effects (p ≤ 0.05) were observed in case of number of leaves (NF), fresh and dry weights of shoot, root and total.