Agriculture has changed dramatically due to mechanization, new technologies, and the increased use of chemical fertilizers. These factors maximize production and reduce food prices, but may also enhance soil degradation. Sustainable agricultural practices include altering crop varieties and the use of soil amendments to increase production, improve irrigation, and more effectively use fertilizers. Ancient and modern durum wheat varieties have been shown to be tolerant to conditions caused by climate change and increase production. Biochar soil amendments have been reported to increase crop yields, soil fertility, and to promote plant growth. However, results are variable depending on biomass source, application conditions, and crop species. This study evaluates the crop response of two contrasting durum wheat varieties on an Eutric Cambisol amended with beech wood biochar. Wheat varieties used are Saragolla, an ancient variety traditionally used in Southern Italy, and Svevo, a widely used commercial variety. The effect of biochar soil amendment on the expression of genes involved in the germination of these two varieties of wheat was determined using RT-PCR. The content of hormones such as gibberellins (GAs), auxins (IAA), and abscisic acid (ABA) was determined. Results demonstrate that biochar had a stimulatory effect on the growth performances of Svevo and Saragolla cultivars at the molecular level. This correlated to the promoted transcription of genes involved in the control of plant development. Overall, the presence of biochar as soil amendment improved the germination rates of both varieties, but the ancient wheat cultivar was better suited to the Eutric Cambisol than the commercial variety. This trend was also observed in un-amended pots, which may indicate better adaptability of the ancient wheat cultivar to withstand environmental stress than the commercial variety.