Computational Molecular Biology 2025, Vol.15, No.5, 227-234 http://bioscipublisher.com/index.php/cmb 230 SPL, ARF, etc., which are responsible for regulating key links such as root development, hormone response, and ROS clearance. For instance, miR408a can affect the accumulation of ROS, and the result might be a negative regulatory effect. When miR164 pairs with MYB or NAC, it plays a role in promoting drought response in the ABA pathway (Figure 1) (Liu et al., 2019; Jiao et al., 2022). The clarification of these regulatory modules not only enables us to understand what they have done, but also provides a clear "bulltarget" for the future cultivation of drought-resistant varieties. Figure 1 The morphology and physiochemical changes of maize variety “M8186” under drought stress. Values followed by different lowercase letters represent p≤ 0.05 (Adopted from Jiao et al., 2022) 5 Construction of miRNA Regulatory Networks and Systems Biology Analysis 5.1 Methods for constructing miRNA-target gene regulatory networks When studying the drought response of corn, it is impossible to obtain a comprehensive picture by relying solely on a single miRNA or target gene. So, putting them on the Internet to look at "relationships" has become a common practice nowadays. Building this network is not entirely dependent on imagination; it requires a three-pronged approach of sequencing data, algorithmic prediction, and experimental verification. For instance, which mirnas showed significant expression changes after drought treatment were screened through tools such as psRNATarget and psRobot, and then degradation omics sequencing was used to verify whether they actually regulated the target genes (Aravind et al., 2017). Next, use software like Cytoscape to graph these interactive relationships, making it clear who controls and who regulates. Furthermore, some people will use WGCNA to analyze which mirnas and target genes are "clustered" in expression, which can dig out potential "core modules" (Tang et al., 2022). 5.2 Connections with ABA, ROS, transcription factors, and other signaling pathways Under drought stress, mirnas do not play a solo role; they are almost always involved in complex networks such as hormones, REDOX reactions, and transcriptional regulation. For instance, the examples of miR164 regulating MYB and NAC transcription factors are often mentioned, and these happen to be involved in ABA-dependent drought responses (Liu et al., 2019; Jiao et al., 2022). For instance, miR408a, during drought, controls the
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