Abstract:Since soybean (Glycine max L. (Merr.)) yields greater than 6719 kg ha −1 have only recently and infrequently been achieved, little is known about the soil microbiological environment related to high-yield soybean production. Soil microbiological properties are often overlooked when assessing agronomic practices for optimal production. Therefore, a greater understanding is needed regarding how soil biological properties may differ between high-and average-yielding areas within fields. The objectives of this study were to (i) evaluate the effects of region on soil microbial carbon substrate utilization differences between high-(HY) and average-yield (AY) areas and (ii) assess the effect of yield area on selected microbiological property differences. Replicate soil samples were collected from the 0-10 cm depth from yield-contest-entered fields in close proximity that had both a HY and an AY area. Samples were collected immediately prior to or just after soybean harvest in 2014 and 2015 from each of seven geographic regions within Arkansas. Averaged across yield area, community-level carbon substrate utilization and Shannon's and Simpson's functional diversity and evenness were greater (p < 0.05) in Region 7 than all other regions. Averaged across regions, Shannon's functional diversity and evenness were greater (p < 0.05) in HY than in AY areas. Principal component analysis demonstrated that a greater variety of carbon substrates were used in HY than AY areas. These results may help producers understand the soil microbiological environment in their own fields that contribute to or hinder achieving high-yielding soybeans; however, additional parameters may need to be assessed for a more comprehensive understanding of the soil environment that is associated with high-yielding soybean.