In order to explore the effect of chemical and Nano bio-fertilizer on quality and quantity of sesame genotypes, an experiment was conducted in a completely randomized block design with three replications at Agricultural Research Institute of Zabol University in 2014-2015 growing season. Treatments included three sesame genotypes (Ardakan, Isfahan and Sistan landrace) as main plot and fertilizer type included 100% of recommended chemical fertilizer, 75% of recommended chemical fertilizer + 1.5 kg.ha^-1 of nano bio-fertilizer, 50% of recommended chemical fertilizer + 1.5 kg.ha^-1 of nano bio-fertilizer, 25% of recommended chemical fertilizer+ 1.5 kg.ha^-1 of nano bio-fertilizer and 1.5 kg.ha^-1 of nano bio-fertilizer as subplots. The results showed that using 100% of recommended chemical fertilizer caused significant increase in plant height, number of capsule per square meter, thousand seed weight, seed yield, and biomass. The highest of capsule number per square meter (2256 number) was obtained from Sistan landrace with the application of 100% of recommended chemical fertilizer. The number of capsule per square meter, number of seeds per capsule, thousand seed weight, seed yield, biomass, harvest index, chlorophyll index and protein percentage showed significant increase in Sistan landrace in comparison with the other genotypes. The results showed that the highest seed yield was obtained (1428.07 kg.h^-1) from Sistan landrace and the highest oil percentage (41.7%) was obtained from Isfahan genotype. According to the results, it could be said that the 100 kg.ha^-1 of potassium sulfate, ammonium phosphate and urea application can be beneficial in increasing yield and improving growth characteristics of sesame.