. Indole-3-acetic acid (IAA) is the type of auxin produced by plants as we l as endophytes present inside the plants. Bacteria do not utilize this hormone but IAA is essential for leaf, root initiation & development of lateral roots. In this study we used four bacterial endophytes Enterobacter asburiae (S2), Bacillus licheniformis (S4), Achromobacter mucicolens (S5) and Pseudomonas fulva (S8) which were isolated from germinating wheat (Triticum aestivum) and screened for plant growth promotional activity in our previous study. Now, these four strains were investigated for production and optimization of Indole-3-Acetic Acid (IAA). Initially IAA production was compared in Luria Broth (LB) and Yeast Extract Mannitol (YEM) media with 0.1% L-tryptophan, YEM media showed maximum production of IAA. To increase productivity of IAA, optimization studies were performed by varying different parameters like, incubation time, carbon source and substrate concentration (tryptophan). Based on the optimization study, yeast extract sucrose media (YES) supplemented 0.2% tryptophan showed higher amount of IAA production after 48 hrs of incubation. Strains Enterobacter asburiae (S2) and Achromobacter mucicolens (S5) gave yield of 251.5 µg/ml and 199.77 µg/ml of IAA respectively. Primary confirmation of IAA was done by thin layer chromatography and both the strains gave good color band & Rf value of samples were compared with standard IAA. FT-IR analysis and LC-ESI-MS of the samples further confirmed the presence of IAA. To check the effect of IAA on plant growth seed germina t ion study were performed. IAA produced by Enterobacter asburiae and Achromobacter mucicolens was checked on Triticum aestivum and Vigna radiata, where significant increase after 21 days in test samples was seen when compared with control. Result indicated that Enterobacter asburiae (S2) showed 68% increase in shoot length while Achromobacter mucicolens (S5) 90% increase in root system. This is a first report that reveals that endophytes of wheat Enterobacter asburiae and Achromobacter mucicolens are indole -3-acetic acid producers and that the IAA produced can be harnessed for increasing root system as we l as healthy growth of other plant.