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Abstract Drought is a major abiotic stress limiting agricultural productivity. Previous field-level experiments have demonstrated that drought decreases microbiome diversity in the root and rhizosphere. How these changes ultimately affect plant health remains elusive. Toward this end, we combined reductionist, transitional and ecological approaches, applied to the staple cereal crop sorghum to identify key root-associated microbes that robustly affect drought-stressed plant phenotypes. Fifty-three Arabidopsis-associated bacteria were applied to sorghum seeds and their effect on root growth was monitored. Two Arthrobacter strains caused root growth inhibition (RGI) in Arabidopsis and sorghum. In the context of synthetic communities, Variovorax strains were able to protect plants from Arthrobacter-caused RGI. As a transitional system, high-throughput phenotyping was used to test the synthetic communities. During drought stress, plants colonized by Arthrobacter had reduced growth and leaf water content. Plants colonized by both Arthrobacter and Variovorax performed as well or better than control plants. In parallel, we performed a field trial wherein sorghum was evaluated across drought conditions. By incorporating data on soil properties into the microbiome analysis, we accounted for experimental noise with a novel method and were able to observe the negative correlation between the abundance of Arthrobacter and plant growth. Having validated this approach, we cross-referenced datasets from the high-throughput phenotyping and field experiments and report a list of bacteria with high confidence that positively associated with plant growth under drought stress. In conclusion, a three-tiered experimental system successfully spanned the lab-to-field gap and identified beneficial and deleterious bacterial strains for sorghum under drought.more » « less
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Hoyer, J. Steen; Pruneda‐Paz, Jose L.; Breton, Ghislain; Hassert, Mariah A.; Holcomb, Emily E.; Fowler, Halley; Bauer, Kaylyn M.; Mreen, Jacob; Kay, Steve A.; Carrington, James C. (, Plant Direct)Abstract ARGONAUTES are the central effector proteins ofRNAsilencing which bind target transcripts in a smallRNA‐guided manner.Arabidopsis thalianahas 10ARGONAUTE(AGO) genes, with specialized roles inRNA‐directedDNAmethylation, post‐transcriptional gene silencing, and antiviral defense. To better understand specialization amongAGOgenes at the level of transcriptional regulation we tested a library of 1497 transcription factors for binding to the promoters ofAGO1,AGO10, andAGO7using yeast 1‐hybrid assays. A ranked list of candidateDNA‐bindingTFs revealed binding of theAGO7promoter by a number of proteins in two families: the miR156‐regulatedSPLfamily and the miR319‐regulatedTCPfamily, both of which have roles in developmental timing and leaf morphology. Possible functions forSPLandTCPbinding are unclear: we showed that these binding sites are not required for the polar expression pattern ofAGO7, nor for the function ofAGO7in leaf shape. NormalAGO7transcription levels and function appear to depend instead on an adjacent 124‐bp region. Progress in understanding the structure of this promoter may aid efforts to understand how the conservedAGO7‐triggeredTAS3pathway functions in timing and polarity.more » « less
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