This study was conducted in order to investigation the reaction of barley root characteristic influenced by using both mycorrhizae and PGPR in Research field of Islamic Azad University of Arak during winter of 2010-2011. Experiment was conducted in a randomized complete block design with three replications. The factors were 1. Azotobacter (two levels included using and not using bacteria), 2. Pseudomonas (three levels included using P. putida, P. fluorescens and not using bacteria) 3. Mycorrhizae (two levels included using and not using fungi). The results indicated that PGPR had a significant effect on Auxin, Cytokenin, GA, protein and phosphor compared to control. The interaction of Mycorrhizae and Pseudomonas had a significant (p<0.01) increase on colonization, biological grain and consecration Zn and Fe, but interaction of Mycorrhizae and Azotobacter were most effective on Zn, Fe concentrations. The results proved the positive effects of microorganism symbiosis with barley root and increase water and nutrition absorption.