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Waterlogging of plants leads to low oxygen levels (hypoxia) in the roots and causes a metabolic switch from aerobic respiration to anaerobic fermentation that results in rapid changes in gene transcription and protein synthesis. Our research seeks to characterize the gene regulatory networks associated with short-term waterlogging. MicroRNAs (miRNAs) are small non-coding RNAs that regulate many genes involved in growth, development and various biotic and abiotic responses. To characterize the involvement of miRNAs in response to hypoxia conditions, a quantitative reverse transcriptase PCR (qRT-PCR) assay was used to profile the expression changes of the 22 candidate mature miRNAs, which were selected from small RNA library of waterlogging resistant line Hz32 and 84 of their predicted targets in three inbred Zea mays lines that showed different tolerance to waterlogging. Based on our studies, miR164, miR167, miR172, miR408 and miR528, which are involved in aerenchyma and root cap formation, lateral root development, root/shoot elongation and plant cell detoxification, found to be key regulator under short-term waterlogging conditions in three inbred lines. Further, computational approaches were used to predict the stress-related cis-regulatory elements on the promoters of these miRNAs using the B73 reference and a miRNA-mediated gene regulatory network was constructed. The differential expression patterns of miRNAs and their targets in these three inbred lines suggest that the miRNAs are active participants in the signal transduction at the early stage of hypoxia conditions via a complex network and biochemical pathways. Our study also suggests that there might be a miRNA-mediated energy-saving strategy in surviving from waterlogging. Examination of miRNA expression in maize root under 4h waterlogging treatment

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