(1) Background: The commercial sustainability of the citrus nursery industry involves cutting costs by using alternative planting substrates to replace (or partially replace) the conventional black peat. Conifer wood biochar was evaluated as a component of the growth medium in a commercial citrus nursery for Carrizo citrange seedlings. (2) Methods: Seven growth media mixtures (A–G) were tested. Each mixture consisted of 50% sandy volcanic soil with the remaining 50% made up as follows: A = black peat and perlite 1:1; B = biochar 1; C = black peat, perlite, and biochar 0.5:1:0.5; D = black peat and biochar 1:1; E = black peat, compost, and biochar 0.5:0.5:1; F = black peat, perlite, compost, and biochar 0.5:0.5:0.5:0.5; G = black peat and lapillus 1:1, this substrate, previously adopted by the hosting nursery, was the control. (3) Results: The best media for the rootstock studied here were those containing 25% biochar (mixtures D and E). In the deeper layers the substrate was more compact, and the roots were limited to the shallow layers of the pot. (4) Conclusions: Conifer wood biochar can be partly added in place of peat in growth media mixtures, thus reducing costs and ameliorating sustainability.