To investigate the effects of organic and chemical fertilizers and their combination on the characteristics of henna (Lawsonia inermis L.) in Southern Kerman province, a factorial farm experiment was conducted in a randomized complete block design with three replications for two years. The factors included rotten cattle manure at two levels (0 (a1) and 15 (a2) ton.ha-1) and NPK fertilizer (urea, triple superphosphate and potassium sulfate) at five levels (0 (b1), 25% (b2), 50% (b3), 75% (b4), and 100% (b5) of the soil test basis). At the end of each year, leaf yield and nitrogen, phosphorus, potassium, and lawson concentrations of the leaves were measured in the different treatments. The results of two-year combined analysis showed that the interaction between organic and chemical fertilizer treatments was significant on yield and leaf lawson concentration. The highest leaf dry weight (483.2 g.m-2) was related to the a2b5 treatment which was not significantly different from a2b4 (473.7 g.m-2). The a2b5 and a2b4 treatments had 41.2% and 38.5% better yield than the control, respectively. Also, the a2b5 treatment showed the highest leaf lawson content (30.1 g.plant-1) without significant difference with a2b4 (25.2 g.plant-1). Considering the situation of the study region in terms of non-use of organic fertilizers and also the economic justification of the produced henna, it could be recommended to use 15 ton.ha-1 cattle manure with a 25% reduction of chemical fertilizer NPK for the henna production.