Parasitism by the root-parasitic plant, Striga (Striga hermonthica L.), is a main threat to pearl millet production in sub-Saharan Africa and nutrient deficiency aggravates this problem, often leading to complete failure of pearl millet crops. Like many other species, pearl millet secretes germination stimulants (strigolactones) into the soil in response to mineral nutrient deficiency, which triggers Striga seed germination resulting in infection. A greenhouse experiment was conducted to evaluate the influence of different doses of di-ammonium phosphate (DAP) fertilizer on strigolactone production and Striga infection in three different African pearl millet cultivars (KBH, Sadore Local and Striga resistance). All the pearl millet genotypes produced varying amounts of different strigolactones like orobanchol, epi-orobanchol, orobanchyl acetate and 5-deoxystrigol, the level of which decreases with increasing doses of DAP. The control treatment (no DAP) showed maximum Striga germination, emergence and dry biomass production in all cultivars of pearl millet. Supply of DAP fertilizer up to 4 g per hill suppressed Striga germination by 69, 64 and 59%; emergence by 87, 85 and 95% and dry biomass by 91, 98 and 83% in cvs KBH, Sadore Local and Striga Resistance, respectively. The present findings reveal that DAP fertilizer minimizes strigolactones production and, as a result, reduces Striga infection in pearl millet. Low doses of DAP fertilizer is a promising strategy to lower the destructive effect of Striga on pearl millet. The use of small doses of DAP fertilizer combined with resistant crop cultivars, intercropping with legumes and hand pulling of Striga at flowering in an integrated Striga control strategy should be developed to help African farmers control this noxious weed.