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Considering current global climate change, drought stress is regarded as a major problem negatively impacting the growth of soybeans, particularly at the critical stages R3 (early pod) and R5 (seed development). Microbial inoculation is regarded as an ecologically friendly and low-cost-effective strategy for helping soybean plants withstand drought stress. The present study aimed to isolate newly drought-tolerant bacteria from native soil and evaluated their potential for producing growth-promoting substances as well as understanding how these isolated bacteria along with arbuscular mycorrhizal fungi (AMF) could mitigate drought stress in soybean plants at critical growth stages in a field experiment. In this study, 30 Bradyrhizobium isolates and 30 rhizobacterial isolates were isolated from the soybean nodules and rhizosphere, respectively. Polyethylene glycol (PEG) 6000 was used for evaluating their tolerance to drought, and then the production of growth promotion substances was evaluated under both without/with PEG. The most effective isolates (DTB4 and DTR30) were identified genetically using 16S rRNA gene. A field experiment was conducted to study the impact of inoculation with DTB4 and DTR30 along with AMF (Glomus clarum, Funneliformis mosseae, and Gigaspora margarita) on the growth and yield of drought-stressed soybeans. Our results showed that the bioinoculant applications improved the growth traits (shoot length, root length, leaf area, and dry weight), chlorophyll content, nutrient content (N, P, and K), nodulation, and yield components (pods number, seeds weight, and grain yield) of soybean plants under drought stress (p ≤ 0.05). Moreover, proline contents were decreased due to the bioinoculant applications under drought when compared to uninoculated treatments. As well as the count of bacteria, mycorrhizal colonization indices, and the activity of soil enzymes (dehydrogenase and phosphatase) were enhanced in the soybean rhizosphere under drought stress. This study’s findings imply that using a mixture of bioinoculants may help soybean plants withstand drought stress, particularly during critical growth stages, and that soybean growth, productivity, and soil microbial activity were improved under drought stress.

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