Abstract
Climate change significantly impedes agricultural growth, development, and production. Plants adapt to environmental changes via the plasticity given by essential genes, which are regulated at the post-/transcriptional level. Gene regulation in plants is a complex process governed by various cellular entities, including transcription factors, epigenetic regulators, and non-coding RNAs. Successful studies have confirmed the function of epigenetic alterations such as DNA methylation/histone modification) in gene expression. In recent years, a highly specialized science known as “Epitranscriptomics” has emerged. Epitranscriptomics studies post-transcriptional RNA chemical alterations seen in all living organisms that alter RNA’s structural, functional, and biological properties. Our minireview interpreted about understanding the molecular pathways: RNA changes and stress-responsive gene regulation. Additionally, the interplay between epitranscriptomics and other regulatory levels has been addressed. In addition, we reviewed technical breakthroughs in epitranscriptomic research, including tools and techniques.
1 Introduction
Evolutionary and ecological constraints impact genome expression via complex regulatory networks. Some primary players governing gene expression dynamics include epigenetic alterations and chromatin re-modeling. The bulk of the previous five decades have seen the sluggish identification of RNA modifications owing to a lack of accurate detection technologies (). However, their finding gained momentum as molecular biology methods, next-generation sequencing technologies, and bioinformatic tools improved, making it feasible to correctly detect transcriptome-wide RNA modifications (). RNA modifications, including the N6-methyladenosine (m6A), offer an additional method for gene regulation, allowing plants to respond more efficiently to environmental changes by precisely modulating gene expression under stressful circumstances. These advancements sparked some fascinating research in RNA biology, culminating in the coining of the phrases “RNA epigenome” and “epitranscriptome”. RNA modification databases have catalogued about 170 chemical modifications (). There is emerging evidence that the epitranscriptome adds a layer to the gene regulation network for plant growth and stress responses ().
Plant hormones are signaling chemicals that play important roles in growth, development, and environmental stress responses. Plant hormones such as abscisic acid (ABA), auxin, brassinosteroid, cytokinin, ethylene, gibberellin, jasmonate, salicylic acid, and strigolactone are well-known plant growth regulators that mediate environmental adaptations (). RNA modifications and plant hormone signals together regulate stress responses by modulating gene expression, ensuring the appropriate genes are activated during adverse conditions. Abiotic stressors such as drought, salt, heat, and floods are becoming more difficult for plants as the climate changes. Abiotic plant stressors reduce energy supply by limiting photosynthesis and energy-releasing catabolic processes (). Climate change and abiotic factors may exacerbate plant diseases (). Such frightening circumstances need novel methods. Recent developments in plant biology have provided critical new insights into how plants detect and react to abiotic stressors. Core stress-signaling pathways include protein kinases similar to yeast sucrose non-fermenting 1 (SNF1) and mammalian AMP-activated protein kinase (AMPK), indicating stress signaling in plants developed from energy sensing (). Stress signaling controls proteins involved in ion and water transport, metabolism, and gene expression reprogramming to achieve ionic and water balance and cellular stability under stressful circumstances (). Stress sensing is typically likened to ligand perception hence it is commonly assumed to take place on the cell surface or at the membrane. The signal would then be sent to multiple subcellular sites, including the nucleus (). Under salt stress, the receptor-like kinase FERONIA may detect salinity-induced cell wall defects and generate cell-specific [Ca2+]cyt signals to preserve cell wall integrity (). A hydraulic signal leads to a quick root-to-shoot water deprivation signal, which triggers ABA production in Arabidopsis leaves and stomatal closure (). Changes in gene expression mediate phytohormones’ effects. Recently, mRNA decay has emerged as another mechanism that contributes to abiotic stress responses (). mRNA decapping is the process by which mRNA molecules degrade. This process involves modifications in RNA, namely m6A. These modifications can designate certain RNA molecules for degradation, hence regulating their stability and the expression of stress-related genes. Recent research indicated that the NAD+-capped transcriptome undergoes considerable alterations in response to ABA and that many ABA-induced transcripts lose their NAD+ caps after ABA treatment, perhaps boosting their stability ().
Transcriptional induction and repression are slower and endure longer than post-translational reactions involving pre-existing proteins, which occur within seconds (). The post-transcriptional regulation level may be best for transitory stress adaption and recovery, which generally take minutes. Quantitative metrics of RNA stability are among the hardest to develop in post-transcriptional regulation. A regulatable promoter may regulate single genes and quantify degradation following transcriptional shut-off (). Plants use post-transcriptional regulation events as an understudied upfront regulatory switch to defend against environmental stress. Multiple constitutively present proteins are activated or deactivated by posttranslational modifications (PTMs) induced by one or more stimuli. Oxidative stress affects protein phosphorylation, sumoylation, ubiquitinylation, cysteine oxidation, glutathionylation, and methionine oxidation ().
We hypothesize that RNA modifications, particularly m6A and NAD+-capping, act as dynamic regulatory switches that integrate environmental signals with post-transcriptional gene regulation, enabling rapid and precise stress responses in plants. Our mini-review focused on epitranscriptomics and stress resistance: a paradigm shift. We also discussed understanding the molecular pathways: RNA alterations and stress-responsive gene regulation. Furthermore, the relationship between epitranscriptomics and various regulatory levels has been explored. Moreover, we examined technical developments in epitranscriptomic research, including tools and approaches.
2 Epitranscriptomics and stress resistance: a paradigm shift
2.1 Types of RNA modifications and their roles in stress responses
RNA mutations regulate plant responses to heat, drought, salinity, and cold. The m6A, 5-methylcytosine (m5C), 7-methylguanosine (m7G), and pseudouridine (P) are important for gene expression and plant life (). The change must be enforced and controlled. The m6 mutation is one of the most studied plant RNA mutations. It activates METTL3, METTL14, and other methyltransferases by cleaving RNA adenine residues (). The m6A alterations have a crucial role in regulating several epigenetic processes. They influence chromatin remodeling, histone modification, and DNA methylation (). For instance, m6A methylation facilitates the recruitment of RNA-binding proteins, including YTH-domain proteins, which regulate the processing, stability, and degradation of stress-related RNA (Figure 1). Moreover, m6A modifications can affect the accessibility of transcription factors to RNA by modifying its secondary structure, referred to as the “m6A switch.” This facilitates the regulation of gene expression in response to stress (). The M6A mutation affects the stability, translation, and splicing of RNA. In response to heat and drought, M6A controls the expression of stress-responsive genes (). During heat stress in plants, m6A methylation speeds up mRNA degradation and dictates the stress response pathways to cope with high temperatures (). These modifications also influence the RNAs associated with chromatin, facilitating the organization of chromatin structure and ensuring the regulation of critical stress-related genes following environmental conditions. Furthermore, METTL3 and METTL14 collaborate with proteins that alter histones and reorganize chromatin, establishing feedback loops that meticulously regulate gene expression during extended stress durations (). This adaptable regulation enables plants to maintain equilibrium and stability even under adverse conditions.
Figure 1
In addition, M6A also affects nuclear transfer RNA, ensuring that transcripts linked to stress are converted into proteins (
Plants adjust m7G levels to adapt to unfavorable conditions during heat and drought efficiently translating stress genes (
2.2 Comparative analysis of epitranscriptomic modifications under different abiotic stresses
Epitranscriptomics is critical for understanding chemical changes in RNA molecules and how plants react to environmental stresses such as drought, heat, cold, and salt. Transcripted RNAs such as m6A, m5C, m7G, and Ψ help control plant gene expression and encourage proper stress responses. A study evaluating these alterations under various stressors demonstrates both typical and common stress response patterns.
(
In cold temperatures, M6A methylation helps the body cope with unfavorable conditions by increasing the resistance of cells. The cleaves of cytosine residues by m5C is another important mechanism for RNA synthesis and modification. In Arabidopsis and rice mutants, m5C contents are affected by heat and drought stress and are genetically influenced (
Besides, pseudouridine (Ψ) alters RNA structure, gene integrity, and transcription may be degraded. The increase in Ψ produces protective proteins, indicating that they work under stress (
3 Deciphering the mechanistic pathways: RNA modifications and stress-responsive gene regulation
3.1 How RNA methylation impacts key stress-responsive genes
Stress genes are regulated by RNA methylation; m6A and m5C are particularly affected during stress.
These changes influence the RNA stability, transcription, splicing, and degradation influence RNA stability transcription, splicing, and degradation influence RNA stability, transcription, splicing, and degradation, helping plants cope with different environmental stresses. Plant RNA changes, such as m6A methylation, are frequent and well-known. A methyltransferase complex triggers it, including METTL3, METTL14, and some other proteins. In m6A mutant, gene expression is greatly influenced by RNA stability and translation efficiency. In heat stress, M6A methylation causes the loss of neutral mRNAs, thus decreasing protein synthesis and enhancing the stress gene transcription involving heat shock proteins (
3.2 Interaction between epitranscriptomic marks and transcription factors in stress adaptation
Stress-induced alterations in plants are regulated by transcripts and signals from epitranscriptomes. Together, epitranscriptomics, which controls RNA molecules, and transcriptomics, which modulates gene expression, create a network that allows plants to resist abiotic conditions such as heat, cold, salt, drought, etc. By turning stress-responsive genes on or off at the appropriate moment, this link enables plants to survive in hostile surroundings. mRNA stability, synthesis, translation, and decrease, therefore impact cell stability via means of epitranscriptomic signals, including m6A, m5C, and m7G.
Among the many stress responses, including cold, drought, and heat, the plant RNA regulator m6A regulates M6A methylation, which modulates mRNA stability and transcription, impacting stress-related genes. By increasing the degradation of redundant mRNAs and creating HSPs, M6A methylation lowers cell damage during heat stress (
Plant stress results from TFs turning genes on or off. It has been shown that RNA-binding proteins alter RNA interactions, influencing TF-epitope interactions. YTHDF (a YTH family protein) and other RNA-binding proteins identify M6A-modified RNA, which then interacts with transcription factors to influence genes (
4 Cross-talk between epitranscriptomics and other regulatory layers
4.1 Integration with miRNA and non-coding RNA networks in stress responses
The stress responses are partly controlled by networks of non-coding RNAs (ncRNAs) and epitranscriptomic miRNA-induced changes. There are several RNA modifications, such as m6A and m5C, which modify the activity of long non-coding ribonucleic acid (lncRNA), microRNAs (miRNA), and other ncRNAs involved in gene regulation (
On the other hand, thermal stress for m6A mutant lncRNA encourages the expression of abnormal proteins among other stress-related genes through mRNA and miRNA (
4.2 Potential interactions with epigenetic modifications and signaling pathways
The interaction between the epitranscriptome and the epigenetic modifications is vital to the regulation layer of plant stress responses. These two regulatory strategies act on the integrated stress response via acting on the transcription level acting on the transcription level, RNA post-transcriptional modification, and translation. They are also related to several signaling pathways that determine how plants perceive and respond to various environmental adversities, e.g., heat, drought, and salt. Methylation of adenosine at position N6 and methylation of cytosine at position C5, which are examples of epitranscriptomic modifications, affect the structure, stability, and translation of RNA molecules, while modifications such as DNA methylation, histone modifications, and chromatin remodeling are epigenetic modifications (
Alterations in chromatin structure, such as histone acetylation and methylation, can drive genes into states where they become more responsive to stress at a given time or are no longer fit for the transcriptional machinery. For instance, heat stress activates the nucleus, which later leads to the appearance of various stress-shock proteins as well as HSPs – proteins involved in heat stress management. It was demonstrated that m6A mutations impact HSF transcript stability as well as translation rates, initiating a quicker stress response (
5 Technological advances in epitranscriptomic research: tools and approaches
Collaboration between epitranscriptomic alterations and epigenetics is critical in plant stress control.
Two levels of regulation mediate stress-induced responses by influencing gene expression at three levels: transcriptional, post-translational, and translation, also involving various signaling pathways. That controls how plants sense and respond to environmental stresses such as heat, drought, and salinity. Epitranscriptomic modifications such as m6A and m5C affect RNA function, stability, and translation, while epigenetic modifications such as DNA methylation histone modifications and deciphering the regulation of chromatin remodeling. The interaction of these two regulatory levels positively affects gene expression, causing it to increase. For example, in drought-prone species, DNA methylation of promoter regions reduces gene expression. However, m6A changes in the mRNA of the same gene enhance or regulate translation. This leads to a rapid stress response (
Histone modifications such as acetylation and methylation can change the structure of chromatin. This makes the gene less sensitive to the transcription mechanism, while m6A can increase or decrease the stability of the mRNA produced by the gene. The radioactivity that controls different stress responses is linked with these changes. For instance, the HSF signaling pathway is activated by heat stress, which provokes the transcription of the HSPs responsible for protein folding. Previous studies have found that the stability and localization of HSF are greatly affected by m6A mutations, causing a quick and efficient response (
6 Future directions: harnessing epitranscriptomics for crop improvement
Epitranscriptomics has great enormous potential for plant evolution. This is particularly important in reproduction and in regulating environmental stresses such as heat, cold, drought, salt and nutritional deficiency. The role of RNA transcripts is an important future direction in stress-induced genes. The identification of specific RNA mutations such as m6A, m5C, and m7G and their role in stress regulation raises the possibility of using these markers to increase plant stress tolerance, such as the m6A mutation, which has been shown to affect the translation and stability of stress-related genes. Additionally, to manipulate RNA editing enzymes, gene editing tools such as CRISPR/Cas9 offer excellent opportunities to achieve desired results. The scientists created high-stress tolerance plants by inhibiting methylate or demethylate RNA enzymes, such as TET for m5C methylation and METTL3 for m6A methylation. This strategy can enhance plant growth under abiotic stress without requiring an incubation period (
Epitranscriptome fits well with other omics techniques such as proteomics, metabolomics, bioinformatics, etc. These techniques could be applied along with epitranscriptome to better understand how plants respond to different stresses. Hence, it provides deep insights into the molecular mechanisms of stress tolerance. The latest bioinformatics tools help researchers in new RNA mutations discoveries and their possible role in stress responses. The important breeding targets have been acquired by developing new RNA-based mutation techniques.
Furthermore, a CRISPR-Cas13-based targeted RNA methylation/demethylation system capable of effectively altering a single mRNA base would considerably expedite research into the relationships between m6A modification and epigenetic regulators (
Artificial intelligence (AI)-powered technologies have been employed to analyze extensive datasets of gene activity to identify genes and networks that react to stress (
Statements
Author contributions
YZ: Conceptualization, Funding acquisition, Project administration, Writing – original draft, Writing – review & editing. AM: Conceptualization, Data curation, Investigation, Validation, Visualization, Writing – original draft, Writing – review & editing. LW: Conceptualization, Writing – original draft, Writing – review & editing. CG: Conceptualization, Writing – original draft, Writing – review & editing. PZ: Conceptualization, Writing – original draft, Writing – review & editing. AE-S: Conceptualization, Writing – original draft, Writing – review & editing.
Funding
The author(s) declare that financial support was received for the research and/or publication of this article. This work was supported by the Foundation of Key Laboratory of Special Waste Water Treatment, Sichuan Province Higher Education System (No. SWWT2024-4) and the research program of the Yibin University (No. 2019QD20).
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Generative AI statement
The author(s) declare that no Generative AI was used in the creation of this manuscript.
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Summary
Keywords
RNA mutations, epigenetics, N6-methyladenosine, environmental stress, stress-responsive genes
Citation
Zeng Y, Muhammad A, Wan L, Gao C, Zhao P and El-Sappah AH (2025) Epitranscriptomic modifications for enhancing abiotic stress resistance in plants. Front. Plant Sci. 16:1538664. doi: 10.3389/fpls.2025.1538664
Received
03 December 2024
Accepted
21 April 2025
Published
23 May 2025
Volume
16 - 2025
Edited by
Wajid Zaman, Yeungnam University, Republic of Korea
Reviewed by
Syed Shams ul Hassan, Shanghai Jiao Tong University, China
Zaid Ulhassan, Zhejiang University, China
Asif Kamal, Quaid-i-Azam University, Pakistan
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Copyright
© 2025 Zeng, Muhammad, Wan, Gao, Zhao and El-Sappah.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Ahmed H. El-Sappah, ahmed_elsappah2006@yahoo.com; 2020110009@yibinu.edu.cn
†These authors have contributed equally to this work and share first authorship
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