This research was carried out to assess bio-fertilizers (Azospirillum brasilense and Pseudomonas fluorescens) for replacing chemical fertilizers to produce organic crop with improvement of Aloe vera root for increasing absorption efficiency micro and macro elements to prevent of ecological effects in the glassy greenhouse in Hassan Abad, Semnan. Aloe vera was planted an offshoot in the pots on 20/6/2009. This research was designed as Randomized Complete Blocks Design (RCBD) in three repetitions with four treatments as follows: (first treatment: control, second treatment: Azospirillum brasilense bacteria, third treatment: Pseudomonas fluorescens bacteria and fourth treatment: integration of A.brasilense and P.fluorescens. Physiological growth parameters were; total fresh weight, shoot fresh weight, root fresh weight, shoot dry weight, root dry weight, length of the biggest leaf, length of the biggest root, leaf number, offshoot number, area of the biggest leaf, plant total area, measuring of Nitrogen, Phosphorus and Potassium elements of the plant in two continuous years. Data were analyzed by using MSTAT-C software and comparison of means by Duncanʼs multi-range test (LSD). The results showed that P.fluorescens treatment vs. control and second treatment (A.brasilense) and fourth treatment (combination of two bacteria) was statistically higher in fresh and dry weight of shoot, number, length, and area of the leaf, plant total area and amount of Nitrogen, phosphorous and potassium elements. A.brasilense treatment vs. control and third treatment (P.fluorescens) and fourth treatment (combination of two bacteria) was statistically higher in total fresh weight and offshoot number. The fourth treatment (integration of two bacteria) vs. control and second treatment (A.brasilense) and third treatment (P.fluorescens) was statistically higher in fresh and dry weight of root and length of the biggest root. The results declare that use of P.fluorescens bio-fertilizer as compared