Abstarct: Nowadays there is an increasingly interest in herbal medicines due to adverse effects of chemical drugs. On the other hand, use of extensive agriculture and innovation of new methods in management of resources utilization for achieving aims of sustainable agriculture have get special importance and hence, use of biofertilizers for reducing in consumption of chemical fertilizers and for increasing of plant yield has became an important challenge for achieving sustainable agriculture. Biofertilizers are composed of different types of free-living microorganisms, having ability to convert basic nutrients from unavailable forms to available ones via biological processes. In this thesis, the effects of vermicompost, arbuscular mycorrhizal fungi and azotobacter on the yield and essential oil of Mentha piperita L. were evaluated without using chemical fertilizers as a split plot on factorial by using of randomized complete block design (RCBD) with three replications. The treatments involved vermicompost as the main factor in six levels including: V0 (no vermicompost), V1 (3 tons of vermicompost enriched per hectare) and V2 (6 tons of vermicompost enriched per hectare) and compositions of arbuscular mycorrhizal fungi and azotobacter as the sub-factors both in two levels where arbuscular mycorrhizal fungi treatments included: M0 (no arbuscular mycorrhizal fungi) and M1 (using arbuscular mycorrhizal fungi) and azotobacter treatments included: A0 (no azotobacter) and A1 (using azotobacter). This research was performed at research farm of Islamic Azad University, Karaj Branch in Mahdasht, Iran. The results showed that vermicompost basic effect on colonization and essential oil percent characteristics has significant relationship in 5% level. Studying the averages shows that using 6 tons of vermicompost enriched per hectare has the best yield between characteristics which have significant relationship in vermicompost factor – 22.58 and 19.25 percent increasing for colonization and essential oil percent respectively compared to the control. Using vermicompost via Improvement in plant yield due to improvement in microbial and biological characteristics of culture medium and regulation of its pH in addition to meaningful increasing in water maintaining capacity may be reason for these findings. Also, there was a significant relationship between the basic effect of arbuscular mycorrhizal fungi for colonization characteristic and the control. This significant relationship, observed only in 1% level caused in 24.19 percent increasing in colonization compared to control. On the other hand, there was a significant relationship in menthon content of essential oil of the plant between azotobacter treatment and control in 5% level; however, unexpectedly the treatment caused in a 32.47 percent decrease in menthon content compared to control. Finally, there were significant relationships between colonization characteristic in arbuscular mycorrhizal fungi and azotobacter treatments in 1% level and also between menthon content of essential oil of the plant in vermicompost and azotobacter treatments in 5% level.