Pine (Pinus sp.) wood products have potential to immobilize fertilizer nitrogen (N) and influence plant growth when used in soilless substrates for the production of containerized floriculture crops. Peat substrate was amended with (by volume) 30% pine wood fiber (peat:fiber) during a production phase with fertigation and a simulated consumer retail phase with clear-water irrigation using container-grown ‘Supertunia Vista Bubblegum’ petunia (Petunia ×hybrida). The objective was to evaluate substrate effects on substrate and plant tissue nutrient level and plant growth, with an emphasis on evaluating N immobilization from wood product amendments. Substrates consisting of peat amended with hammer-milled pine wood (peat:wood) or coconut (Cocos nucifera) coir (peat:coir) were used for comparison, and a 100% peat substrate (peat) served as a control. In Expt. 1, amending peat with pine wood fiber had no effect on leaf SPAD chlorophyll index, shoot growth, plant height and width, substrate N, or percent shoot tissue N at the end-of-production. In Expt. 2, plants grown in peat:fiber had reduced flower number, plant height and width, and shoot growth compared with plants grown in the 100% peat control. However, petunia grown in peat:fiber substrates maintained dark-green foliage with high leaf SPAD chlorophyll index values (≥44.4) and ≥45 flowers/plant, and therefore were considered marketable plants. During the production phase in both Expts. 1 and 2, N concentrations remained within the target range for petunia in both the shoot tissue and root-zone for all substrates. In addition, there was no statistical evidence of N immobilization for any substrate blend for either of the N drawdown procedures. In both Expts. 1 and 2, root-zone nutrients became depleted during the consumer phase when irrigation was with clear water (no fertilizer), and petunia developed uniform symptoms of leaf chlorosis and N deficiency. Results of this study indicate that peat amended with 30% pine wood fiber, hammer-milled pine wood, and coconut can be used for production of containerized petunia with minimal effects on plant growth or need to adjust the fertilizer program. However, increasing pine wood to >30% of the substrate volume may require growers to increase fertilization and adjust irrigation practices to compensate for greater risk of N immobilization and changes in substrate physical properties.