Bioscience Methods 2025, Vol.16, No.6, 280-288 http://bioscipublisher.com/index.php/bm 280 Feature Review Open Access Post-Transcriptional Regulation by microRNAs during Drought in Rye Guiping Zhang, Wei Wang Institute of Life Sciences, Jiyang College of Zhejiang A&F University, Zhuji, 311800, Zhejiang, China Corresponding email: wei.wang@jicat.org Bioscience Methods, 2025, Vol.16, No.6 doi: 10.5376/bm.2025.16.0027 Received: 10 Sep., 2025 Accepted: 26 Oct., 2025 Published: 11 Nov., 2025 Copyright © 2025 Zhang and Wang, This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Zhang G.P., and Wang W., 2025, Post-transcriptional regulation by microRNAs during drought in rye, Bioscience Methods, 16(6): 280-288 (doi: 10.5376/bm.2025.16.0027) Abstract Rye (Secale cereale L.) is renowned for its strong adaptability in marginal environments and is of great significance for ensuring food security and achieving sustainable agriculture in areas with frequent droughts. This study systematically explored the miRNA expression dynamics of rye under drought stress and its mechanism of action in gene silencing, signal integration and stress adaptation. Through high-throughput sequencing technology, multiple differentially expressed mirnas under drought conditions were identified, and their potential target genes were predicted. The results of functional annotation and network analysis indicated that these mirnas were mainly involved in regulating key pathways such as reactive oxygen species (ROS) clearance, abscisic acid (ABA) signaling, and transcriptional regulation related to stress responses. In the case studies of miR398, miR159 and miR166, the research revealed how specific mirNA-target gene interactions affect the physiological characteristics of rye under drought stress, including ROS clearance efficiency, hormone signal regulation and leaf morphology changes, etc. This study not only deepens the understanding of the molecular response mechanism of rye to drought, but also provides new ideas and technical support for mirNa-based functional genomics research and the improvement of stress-resistant crops in the context of climate change. Keywords Rye; Drought stress; microRNA; Post-transcriptional regulation; Drought resistance mechanism 1 Introduction Rye (Secale cereale L.) has not been a leading grain for a long time, but it has shown astonishing resilience in some difficult growing environments, especially in dry or infertile soil conditions, where it is more resilient than most cereal crops. So, as the global climate becomes increasingly extreme, especially with the frequency and intensity of droughts continuing to rise, rye has naturally been placed back on the key list for research and breeding. It not only plays a complementary role in food security, but also offers a relatively safe option for sustainable agriculture. However, to enable rye to perform better under water shortage conditions, it is necessary to figure out what molecular mechanisms are "supporting" it (Cheng et al., 2022). In recent years, researchers have increasingly focused their attention on microRNA (miRNA). These small non-coding RNA molecules, which are only about 20 bases in length, can "dictate" a large number of genes after transcription, and they are extremely precise. They not only involve key drought resistance links such as abscisic acid (ABA) signaling, ROS clearance system and root configuration, but also have been confirmed to be involved in the response regulation under drought stress in a variety of plants (Singh et al., 2022). Although these findings mainly come from well-studied crops such as rice and corn, rye and its "distant relatives" - other gramineous plants - also show signs that the regulatory potential of miRNA should not be underestimated (Ferdous et al., 2015; Singroha et al., 2021; Zhang et al., 2022; Zhakypbek et al., 2025). At present, many studies are no longer merely exploring the existence of these small Rnas, but have gone further, taking them as potential entry points for future crop stress-resistant breeding. This study aims to systematically explore the post-transcriptional regulatory mechanism mediated by miRNA under drought stress in rye, review the current understanding of miRNA biosynthesis and function in plant drought response, summarize the latest research achievements of miRNA in drought response in rye and its related species, and discuss the application of bioinformatics and experimental methods in miRNA target identification and validation. By clarifying the regulatory network related to miRNA, this study aims to provide a scientific
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