<p>The use of microbial inoculant is a cost-effective and eco-friendly strategy to enhance stress tolerance while improving plant growth and development in sustainable agricultural systems. This study aimed to evaluate the potential of <i>Azospirillum</i> strain inoculated to transgenic <i>AVP1</i> wheat plants. Four bacterial strains were isolated from the rhizosphere of wheat and were identified as <i>Azospirillum</i> sp. by colony morphology and 16S rRNA gene sequencing analysis. Production of indole acetic acid (IAA) and gibberellic acid (GA) was determined and quantified in a pure culture. The <i>AVP1</i> (H⁺-pyrophosphatase) gene was transformed into Seher-2006 wheat variety and evaluated for its effect on growth improvement. Results showed that <i>AVP1</i> transgenic plants inoculated with <i>Azospirillum</i> strains S-11y and S-12d1 produced significantly higher biomass and grain yield compared to non-inoculated transgenic plants. Available nitrogen was also higher in the rhizosphere of inoculated transgenic plants. However, phosphatase activity decreased, while dehydrogenase and protease activities increased after inoculation. It can be suggested that root exudates produced by <i>AVP1</i> transgenic plants supported the growth of plant growth-promoting bacteria, and more robust root system was developed. These results indicated that the inoculation of <i>AVP1</i> transgenic plants with plant growth promoting <i>Azospirillum</i> strains has a profound effect on increasing biomass and grain yield. Thus, these potent bacterial strains may also be used as commercial biofertilizers for improving plant growth and yield.</p>