The objective was to quantify the effect of the timing of macronutrient applications on nutrient uptake, growth, and development of Petunia ×hybrida Hort. Vilm.-Andr. ‘Supertunia Royal Velvet’ during vegetative propagation. Starting with unrooted cuttings (Day 0), fertigation was applied continuously at three time intervals (Day 0 to 7, Day 8 to 14, or Day 15 to 21) using either a “complete” (C) water-soluble fertilizer containing (in mg·L−1) 75 NO3-N, 25 NH4-N, 12 phosphorus (P), 83 potassium (K), 20 calcium (Ca), 10 magnesium (Mg), 1.4 sulfur (S), 2 iron (Fe), 1 manganese (Mn), 1 zinc (Zn), 0.5 copper (Cu), 0.5 boron (B), and 0.2 molybdenum (Mo) or a micronutrient fertilizer (M) containing (in mg·L−1) 1.4 S, 2 Fe, 1 Mn, 1 Zn, 0.5 Cu, 0.5 B, and 0.2 Mo in a complete factorial arrangement. With constant fertigation using the C fertilizer, plant dry weight (DW) doubled from Day 0 (sticking of unrooted cuttings) to Day 7 (0.020 g to 0.047 g), root emergence was observed by Day 4, and by Day 7, the average length of primary roots was 2.6 cm. During any week that the M fertilizer was substituted for the C fertilizer, tissue N–P–K concentrations decreased compared with plants receiving the C fertilizer. For example, plants receiving the M fertilizer between Day 0 and 7 had 20% lower tissue-N concentration at Day 7 compared with those receiving the C fertilizer. Although both shoot DW and leaf count increased once macronutrient fertilization was resumed after Day 7, final shoot DW and leaf count were lower than plants receiving C fertilizer from Day 0 to 21. Time to first root emergence was unaffected by fertigation. Constant application of C resulted in a higher shoot-to-root ratio at Day 21 than all other treatments. Results emphasize the importance of early fertigation on petunia, a fast-rooting species, to maintain tissue nutrient levels within recommended ranges.