Next Article in Journal
Comparing Inflammatory Biomarkers in Cardiovascular Disease: Insights from the LURIC Study
Previous Article in Journal
Interactions Between Prolactin, Intracellular Signaling, and Possible Implications in the Contractility and Pathophysiology of Asthma
Previous Article in Special Issue
Artificial Intelligence-Assisted Breeding for Plant Disease Resistance
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
IJMS
Volume 26
Issue 15
10.3390/ijms26157334
ijms-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Review

MicroRNA528 and Its Regulatory Roles in Monocotyledonous Plants

by
Hailin Fu
,
Liwei Zhang
,
Yulin Hu
,
Ziyi Liu
,
Zhenyu Wang
,
Fafu Shen
and
Wei Wang
*
College of Agronomy, Shandong Agricultural University, No. 61 Daizong Street, Tai’an 271018, China
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2025, 26(15), 7334; https://doi.org/10.3390/ijms26157334
Submission received: 24 June 2025 / Revised: 23 July 2025 / Accepted: 28 July 2025 / Published: 29 July 2025
(This article belongs to the Special Issue Latest Reviews in Molecular Plant Science 2025)
Browse Figures
Versions Notes

Abstract

MicroRNA528 (miR528) is a microRNA found only in monocotyledonous (monocot) plants. It has been widely reported that miR528 is involved in the regulation of plant growth and development, such as flowering, architecture, and seed and embryogenic development, in addition to playing a crucial role in response to various biotic and abiotic stresses, such as plant pathogens, salt stress, heat/cold stress, water stress, arsenic stress, oxidative stress, heavy-metal stress, and nutrient stress. Given that it is specific to monocot plants, to which the major staple food crops such as rice and wheat belong, a review of studies investigating its diverse functional roles and underlying mechanisms is presented. This review focuses on the processes in which miR528 and its targets are involved and examines their regulatory relationships with significant participation in plant development and stress responses. It is anticipated that more biological functions and evolutionary effects of miRNA targets will be elucidated with the increase in knowledge of miRNA evolution and examination of target mRNAs.

1. Overview of Role of miRNAs in Plant Development and Stress Responses

MicroRNAs (miRNAs) are a class of single-stranded non-coding small RNAs (sRNAs) with a mature sequence length ranging from 20 to 24 nucleotides [1]. In plants, miRNAs are processed from larger precursor molecules that are transcribed from miRNA genes (MIRs) by RNA polymerase II (Pol II). The precursor molecules form self-complementary stem loop structures and are processed into miRNA:miRNA-star duplexes by a dicing complex, including DICER-LIKE 1 (DCL1), HYPONASTIC LEAVES 1 (HYL1), and SERRATE (SE). The miRNA:miRNA-star duplexes are 2′-O-methylated at the 3′ ends by the methyltransferase HUA ENHANCER 1 (HEN1) within the nucleus and are exported into the cytoplasm, where they are incorporated into RNA-induced silencing complex (RISC) by HASTY (HST), which is a member of the importin β family of nucleocytoplasmic transporters. Upon RISC loading, the mature, methylated miRNA is complexed with Argonaute 1 (AGO1), while the miRNA-star strand is removed and degraded [2]. The mature miRNA functions to convey sequence-specific negative regulation of endogenous or exogenous target mRNAs by degradation or translation repression through sequence complementarity at the post-transcriptional level [3,4].
In the past two decades, miRNAs have been increasingly recognized for their involvement and the pivotal roles they play in regulating plant growth, development, and responsiveness to various environmental stress conditions [5,6,7]. For example, wheat (Triticum aestivum L.) miRNA tae-miR408 was found to determine the heading time by mediating the expression of TIMING OF CAB EXPRESSION-A1/B1/C1 (TaTOC1s), which is known as a key component of the plant circadian clock [8]. It was also demonstrated that tae-miR408 can mediate plant responses to phosphate (Pi) starvation and salt stress by regulating the expression of a suite of six genes that are associated with Pi acquisition and ABA signaling transduction [9]. On the other hand, transgenic expression of short tandem target mimic (STTM), which targets genetic downregulation of 35 miRNA families in rice (Oryza sativa L.), resulted in hereditable alterations in a number of agronomic traits, including plant height, tiller number, and grain number [10]. Among these targeted miRNAs, osa-miR398 plays a crucial role in determining panicle length, grain number, and grain size in rice [10]. In addition, it positively regulates rice basal defense against the blast fungus Magnaporthe oryzae by boosting hydrogen peroxide (H2O2) production through the regulation of CSD1 and CSD2, which encode Cu/Zn superoxide dismutases (SODs), and CCSD, a copper chaperone for SOD [11]. However, osa-miR398 exhibits a negative regulatory role in plant defense in other species, such as Arabidopsis thaliana [12] and barley [13].
A striking feature of miR528 in monocotyledonous plants is the remarkable diversity of its expression patterns and biological functions across different species. Numerous studies have demonstrated that miR528 exhibits species-specific regulatory roles, with some even reporting contrasting expression levels and functional outcomes among different monocots. Despite such variations, miR528, as a monocotyledon-specific microRNA (miRNA), plays a significant regulatory role in plant growth, development, and responses to environmental stresses. For example, miR528 is reportedly involved in plant development processes, such as flowering [14,15], plant architecture [16], and seed and embryogenic development [17], in addition to mediating plant responses to various biotic and abiotic stresses, such as plant pathogens [18], salt stress [16], heat/cold stress [19], water stress [20], arsenic (As) stress [21], oxidative stress [22], heavy-metal stress [23], and nutrient stress [24]. Understanding the miR528-mediated regulatory network would undoubtedly highlight miRNA studies as a fruitful avenue for further investigations that may have far-reaching implications in studying molecular regulation of important agronomic traits and plant defense in monocot crops and provide a set of new gene tools for crop improvements by marker-assisted breeding and biotechnology. Here we attempt to provide a synopsis of the growing recognition that miR528 is a significant player in modulating plant growth, development, and stress responses, imparting a solid knowledge base for further investigations on the more defined biological functions and evolutionary aspects of miR528 and its molecular targets.

2. miR528 and Its Targets

2.1. MIR528 Genes in Monocots

Since its first discovery in rice [25], miR528 has been identified in many other monocot plants, such as wheat [26], maize [27], sorghum [28], and sugar cane [29]. To facilitate the investigations of characteristic features of miR528 loci (MIR528), we extracted the miR528 information from 17 monocot plant species in the Plant miRNA ENcyclopedia (PmiREN; version 2.0) database (http://www.pmiren.com/, accessed on 20 January 2025) [30]. As shown in Table 1, apart from maize (Zea mays L.), which has two members, and switchgrass (Panicum virgatum L.) and wheat (Triticum aestivum L.), which have three members, miR528 is encoded by a single gene. The precursors of miR528 are endowed with a well-developed stem-loop structure (Supplementary Figure S1), and their mature sequences are identical, except for scu-miR528 derived from the Saccharum hybrid cultivar, which differs from the others by a single nucleotide at the 3′ end (Figure 1A).
To explore the evolutionary history of miR528 in monocots, we reconstructed a phylogenetic tree of the miR528 precursors. The presence of MIR528 can be traced back as early as in an Alismatales duckweed species (Spirodela polyrhiza L. Schleid) but not in any eudicots, gymnosperms, or basal angiosperms (Figure 1B and Figure S1). It is plausible that miR528 emerged after the split of monocots from their common ancestor with eudicots, and the phylogenetic relationships of miR528 homologs in each subgroup were consistent with plant speciation and evolution (Figure 1B and Supplementary Figure S1). It has recently been revealed that MIR528 genes were found in a syntenic block of asparagus (Asparagus officinalis L.), pineapple (Ananas comosus L.), and rice, but such a syntenic relationship was lost in banana (Musa acuminata Colla) [19]. In general, our analysis of the updated PmiREN database confirms that miR528 is a miRNA restricted to monocots.

2.2. Targets of miR528

Many studies indicate that the target genes of miR528 may have undergone high levels of divergence [16,18,35]. To further reveal the target genes of miR528, we performed genome-wide search and prediction of miR528 targets in the five grain or forage crop plants with economic significance, including O. sativa, Z. mays, T. aestivum, Setaria italica, and P. virgatum using the plant small RNA target analysis server (psRNATarget) web server [36], in combination with relevant public degradome sequencing data that were gathered from the PmiREN database. As a result, 19, 23, 13, 14, and 22 putative target genes of miR528 were predicted in O. sativa, Z. mays, T. aestivum, Setaria italica, and P. virgatum, respectively (Supplementary Table S2). The putative target genes of miR528 showed both structural conservation and specificity among the five examined plant species.
The conserved genes that are targeted by miR528 encode three types of proteins, including plastocyanin-like domain-containing proteins (PLC), proteins containing the F-box domain and (or) leucine-rich repeat (LRR), and enzymes related to reactive oxygen species (ROS) homeostasis (Supplementary Table S2). The predicted target genes of miR528 in both rice and maize include all the three types of proteins mentioned above (Figure 2B–D), indicating that miR528 has conserved targets across different monocotyledonous species. The ROS homeostasis-related proteins include polyphenol oxidase (PPO), ascorbate oxidase (AAO), amine oxidase (AO), SOD, and peroxidase (POD), which are involved in both ROS generation and scavenging. As both the plastocyanin-like domain-containing proteins and the ROS homeostasis-related proteins are copper-containing proteins, miR528 is also regarded as a copper miRNA (Cu-miRNA) together with miR398, miR397, miR408, and miR857 [37]. The alignment of miR528 target sites in the three types of target genes revealed seven consensus sites that could be attributable to conserved targeting (Figure 2A and Supplementary Figure S2A). This is further corroborated by the analysis of degradome sequencing data, which verified the associations between miR528 and the three types of conserved targets (Figure 2B–D).
While some target genes of miR528 are conserved among monocotyledonous species, others exhibit species specificity and functional diversity. To highlight this aspect, we selected four non-conserved yet functionally significant targets for detailed illustration (Figure 2E–H and Supplementary Table S2). For instance, among the predicted target genes of osa-miR528, LOC_Os08g42640 encodes a putative C3HC4-type zinc-finger domain-containing protein that regulates flowering time (Figure 2E). Likewise, Z. mays zma-miR528 was predicted to target GRMZM2G118312, which encodes a putative galactoside 2-alpha-L-fucosyltransferase (Figure 2F); S. italica sit-miR528 targets a theobromine synthase gene Seita.4G137600 (Figure 2G); T. aestivum tae-miR528 was predicted to target a PLATZ transcription factor, Traes_6AS_3CC6C63F9, which mediates leaf growth and senescence in plants (Figure 2H). The above examples indicate that miR528 targets differ among various monocotyledonous plants, yet all these targets are functionally important. The alignment of all the miR528 target sites led to the identification of one consensus site in P. virgatum; two consensus sites in each of O. sativa, Z. mays, and T. aestivum; and three consensus sites in S. italica (Figure 2A and Supplementary Figure S2B–F).

3. Role of miR528 in Plant Growth and Development

3.1. Flowering Time

At the transcriptional level, osa-miR528 is highly expressed concomitant with plant development and is light-inducible with a diurnal rhythm. At the post-transcriptional level, the transcript level of mature osa-miR528 in rice leaves is fine-tuned by the alternative splicing (AS) and alternative polyadenylation (APA) events of primary osa-miR528 (pri-osa-miR528) [14]. Tissue-specific expression analysis, coupled with 5′ RNA ligase-mediated rapid amplification of cDNA ends (RLM-RACE) analysis, showed that osa-miR528 targets a conserved C3HC4-type zinc-finger transcription factor gene, which is also known as RED AND FARRED INSENSITIVE 2 (OsRFI2). Transgenic overexpression of osa-miR528 was found to attenuate the expression of OsRFI2 and promote early flowering under long-day conditions [14]. Further, the natural variations in the osa-MIR528 promoter led to different expression levels of osa-miR528 in 50 accessions of wild and cultivated rice, which are associated with variations in binding affinities of the SQUAMOSA PROMOTER-BINDING PROTEIN 7 (OsSPL7) transcription factor, which activates osa-MIR528 expression. Consequently, a conserved photoperiodic flowering pathway in rice mediated by the SPL7–miR528–RFI2 module was established, which likely contributes to the geographic adaption of rice to grow at different latitudes, corresponding to different photoperiod rhythms [14]. miR528 has also been proposed to regulate flowering time by modulating secondary metabolism. For instance, in Hemerocallis citrina, it targets the key structural gene PGT1-7 and the transcription factor bZIP-1 to control flavonoid modification and accumulation during the bud-to-flower transition, thereby influencing floral organ development and maturation [39].

3.2. Plant Architecture

It is known that miR528 plays a regulatory role in plant development. Transgenic overexpression of pri-osa-miR528 in creeping bentgrass (Agrostis stolonifera L.), which is an important perennial grass species, led to alterations in plant architecture [16]. Transgenic creeping bentgrass exhibited a distinct dwarf phenotype featuring reduced average length of the internodes from each tiller, thicker leaves, and increased number of vascular bundles and tillers, resulting in a denser, uniform, and lodging-resistant plant architecture, which is highly desired in turf grass [16]. Consistent with this, in Leymus chinensis, the CRISPR-associated protein 9 (CRISPR/Cas9)-mediated knockout of miR528 led to a significant increase in tiller number—on average two more tillers than wild-type plants both 1 and 3 months post-transplantation—resulting in enhanced growth rate and biomass accumulation [40]. Further mechanistic insights come from rice, where miR528 regulates plant architecture by targeting the D3 gene, a key regulator of abscisic acid (ABA) and gibberellin (GA) biosynthesis; its overexpression represses D3 expression, reducing GA and elevating ABA levels, thereby enhancing tillering and reducing plant height by modulating hormonal homeostasis [41].

3.3. Seed Development

In comparison to the aforementioned roles in flowering time and plant architecture, there are more studies that demonstrated the functionality of miR528 in regulating seed development. In maize, the dynamic expression profiling of miRNAs by Solexa deep sequencing in Zhengdan 958, an elite maize hybrid and cultivated widely in China, revealed that the abundance of zma-miR528a and zma-miR528b decreased linearly in the grain filling stage; and eight putative target genes of zma-miR528 were identified, which could be grouped by their diverse functionalities, including oxidoreductase activity, signal transduction, development, post-translational regulation, and stress response [17]. Building on its expression dynamics during seed maturation, subsequent studies further demonstrated that zma-miR528a is transcriptionally activated during seed germination by high nitrate and auxin levels through TGACG motif-binding factors 1 and 4 (TGA1/4) and Auxin Response Element (AuxRE) in its promoter, thereby promoting its accumulation and facilitating seed germination via the regulation of redox homeostasis and hormonal signaling [42]. In the maize inbred line Chang 7–2 (paternal parent of Zhengdan 958), the potential functional role of zma-miR528 could also be predicted by its high abundance in the latter stage of nutrient storage during kernel development [43]. Meanwhile, in the maize inbred line B73, zma-miR528 was highly expressed during seed development [44].
In the rice cultivar Nipponbare, high-throughput sequencing data showed that osa-miR528 was highly expressed in developing rice grains with 7.5-fold greater expression 6–10 days after pollination (DAP) than 1–5 DAP [45]. In the japonica cultivar Zhonghua 11, osa-miR528 was highly and preferentially expressed in embryos [46]. The putative target genes of osa-miR528 have been identified to encode copper-binding proteins and L-ascorbate oxidases (AOs), which are involved in diverse developmental and metabolic processes, such as regulating the apoplastic redox state and modulating plant growth and defense responses [46]. Taken together, these results suggest that miR528 may play a vital role in plant seed development by regulating the expression of a number of target genes.

3.4. Embryogenic Development

During somatic embryogenesis (SE), a high level of miR528 expression is observed in several plant species, such as rice [47] and maize [48,49]. In rice, osa-miR528 shows high expression in undifferentiated calli and maintains the cells in the meristematic phase by targeting genes coding copper-containing oxidase enzymes, which results in lower production of lignin and a thinner cell wall [47,50]. In maize, zma-miR528 is required for embryo dedifferentiation during somatic embryogenesis, and it regulates multiple target mRNAs, including basic helix-loop-helix transcription factors, multidrug and toxic compound extrusion/big embryo, SOD, and PLC, by promoting their degradation, translation inhibition, or both [48,49,51,52].

4. Role of miR528 in Plant Stress Responses

4.1. Biotic Stress

It has been suggested that miR528 may play a role in plant antiviral immune responses by targeting immunity-associated genes [53]. Rice plants that were inoculated with rice stripe virus (RSV) showed reductions in the expression of miR528 and an increase in the accumulation of AGO1 and Argonaute 18 (AGO18) proteins [18]. As a decoy AGO protein incapable of cleaving target RNAs, AGO18 was able to sequester miR528 and prevent it from its incorporation into RNA-induced silencing complex (RISC) with AGO1 [53]. As a result, the reduced RISC efficiency allowed for higher expression of the AO gene that catalyzes ascorbic acid (AsA) oxidation, leading to higher basal ROS accumulation and enhancement in ROS-mediated resistance against RSV infection [18].
In a subsequent study, the same research team reported that the miR528-AO antiviral defense pathway was regulated by the transcription factor SQUAMOSA Promoter-Binding Protein-Like 9 (SPL9), which positively regulated miR528 expression by binding to the GTAC motifs in the MIR528 promoter and negatively regulated the antiviral defense in rice, whereby the transgenic lines overexpressing SPL9 showed exacerbated symptoms and higher infection rates than the wild-type (WT), whereas the rice spl9-knockout lines showed alleviated RSV infection compared with the WT [54]. Building on these findings, a recently proposed regulatory model further describes a complete copper (Cu)–SPL9–miR528–AO–ROS–antiviral pathway in rice, in which virus infection induces copper transporter gene expression and leaf copper accumulation, thereby inhibiting the SPL9-mediated transcriptional activation of miR528, leading to the upregulation of its target gene AO, enhanced ROS production, and ultimately increased antiviral defense capacity [55]. It is, therefore, envisaged that the genetic manipulation of miR528-mediated gene regulation may impart new and effective tactics to enhance pathogen immunity in crop plants.
Beyond viral defense, miR528 also contributes to insect resistance in rice. Specifically, during brown planthopper (BPH) infestation, the long non-coding RNA (lncRNA) MSTRG.13957.30 is upregulated and functions as a target mimic, binding osa-miR528 and preventing it from suppressing the AO gene. This regulatory interaction enhances AO expression and activates ascorbic acid-related defense pathways, thereby contributing to BPH resistance [56].
Furthermore, miR528 has been shown to function in cross-kingdom immune regulation. In maize, zma-miR528b-5p directly targets the fungal virulence gene FvTTP in Fusarium verticillioides, reducing fumonisin B production and promoting salicylic acid (SA) accumulation and pathogenesis-related protein 1 (PR1) expression, ultimately reinforcing SA-mediated immune responses in the host plant [57].

4.2. Salt Stress

The role of miR528 in response to salt stress was investigated in creeping bentgrass where miR528 was significantly induced by salt stress [16], which was corroborated by similar observations in maize [58]. Upon salt treatment, the overexpression of rice osa-miR528 in creeping bentgrass directly repressed the expression of its target genes AAO and copper-ion-binding protein 1 (CBP1), which mediated oxidation homeostasis and alleviated cellular damage by salt stress. It was postulated that miR528 may also positively regulate the genes involved in some other signaling pathways, such as a high-affinity K transporter (HAK5) and a salt stress-induced transcription factor (NAC60), induce the activity of antioxidant enzymes such as catalase (CAT) to maintain ROS homeostasis, and interact with other stress-related miRNAs to form a regulatory network to coordinately integrate multiple stress regulators in response to salt stress [16]. Consistently with these findings, in rice, the salt-induced upregulation of miR528 was shown to suppress the expression of its target gene AO, thereby increasing the accumulation of AsA and ABA, which subsequently promoted proline synthesis, enhanced osmotic adjustment and ROS scavenging, and ultimately improved salt tolerance [59].

4.3. Temperature Stress

Temperature stress is one of the major environmental factors that affects not only plant growth but also fruit postharvest shelf life and quality. Using deep sequencing and bioinformatic and molecular analyses, in banana, the expression of mac-miR528 was severely suppressed by low temperature but significantly induced by heat stress [19]. The opposite effect was observed in switchgrass, where miR528 was significantly attenuated by heat stress [60]. Also, in banana, degradome sequencing analysis revealed that miR528 acts in concert with its corresponding target gene (PPO), which plays an imperative role in regulating plant response to temperature stress [19]. Further supporting these findings, exogenous melatonin treatment upregulates miR528 expression in banana, which represses its target gene MaPPOs, thereby reducing PPO activity, enhancing peroxidase (POD) and CAT activities, maintaining ROS homeostasis, and ultimately improving cold tolerance and delaying peel browning [61]. In rice, the co-overexpression of osa-miR528, osa-miR397, and osa-miR408 synergistically enhances cold tolerance by reducing leaf damage, ion leakage, and malondialdehyde (MDA) accumulation under low-temperature stress [62]. Consistently, in the cold-tolerant japonica rice variety JL (Jilin Sunset), osa-miR528 was significantly downregulated under cold stress, leading to the de-repression of its target gene Os03g0152000, enhanced antioxidant capacity, and improved cold tolerance [63]. MiR528 plays an important regulatory role in plant responses to low-temperature stress by modulating redox balance and maintaining cellular homeostasis. However, its specific regulatory mechanisms remain unclear and warrant further investigation.

4.4. Water Stress

It has been recognized that miR528 could mediate post-transcriptional regulation in response to water stress, which is a major constraint for crop yield improvements. In wild emmer wheat (Triticum turgidum ssp. dicoccoides), the ancestor of domesticated durum wheat (T. turgidum ssp. durum), the expression of miR528 was significantly downregulated in leaf and root tissues by drought stress [20]. Likewise, in sugarcane (Saccharum spp.), ssp-miR528 was downregulated in both sugarcane cultivars RB867515 (high tolerance to drought) and RB855536 (low tolerance to drought) on the fourth day of drought stress treatment, which is suggestive of its role in reducing growth in response to drought stress [64]. When the seedlings of three maize inbred lines (Hz32, B73, and Mo17) with different sensitivities to waterlogging were assayed under controlled experimental conditions, it was found that miR528 modulates root development as a key regulator in the post-transcriptional regulatory system in response to short-term waterlogging stress [65]. Further supporting its functional versatility, the overexpression of osa-miR528-3p in rice enhances drought tolerance by increasing indole-3-acetic acid (IAA) levels and reducing ROS accumulation, thereby promoting root and leaf elongation, as well as biomass accumulation [66]. Furthermore, in Agropyron mongolicum, miR528 is downregulated under drought stress, leading to the upregulation of its target gene HOX24, a transcription factor involved in ABA signaling and water deficit response, thereby suggesting a regulatory role of miR528 in drought tolerance via modulation of HOX24-mediated pathways [67]. In rice, osa-miR528 displays differential expression under drought stress in the jointing stage across varieties, with notably altered levels in the drought-sensitive cultivars IR64 and Kitaake, indicating its potential role as a key regulatory node in mediating reproductive-stage drought responses [68]. In line with these findings, in the drought-tolerant rice variety Azucena, the drought-induced downregulation of miR528 enhances drought tolerance by increasing the activities of antioxidant enzymes such as SOD and peroxidase (POX), reducing ROS accumulation and improving root tip cell viability and elongation capacity [69]. Collectively, these findings indicate that miR528 regulates both hormonal signaling and antioxidant defense pathways, thereby acting as a central hub in plant drought stress responses.

4.5. Arsenic (As) Stress

Arsenic (As), which is ubiquitous and abundant in the earth’s crust, is a nonessential metalloid and very toxic to plants. Arsenite [As (III)] and arsenate [As (V)] are the predominant inorganic species of As in soil and are readily interconvertible depending upon soil changing redox potential and pH [70]. Rice is highly efficient in As uptake, particularly in As-contaminated soils, relative to other cereals, such as barley and wheat [71]. In rice, the analysis of miRNA expression pattern under As (III) stress at the genome-wide level by using a high-throughput sequencing strategy and stem-loop real-time quantitative PCR revealed that osa-miR528 was strongly upregulated by As (III) in the roots of Minghui 86, a rice cultivar that is sensitive to As (III) stress [72]. It is hypothesized that in the proposed network of arsenite-responsive miRNAs, the upregulation of miR528 in the roots of rice seedlings in response to As stress downregulate the copper-ion-binding protein (CBP) and IAA-alanine resistance protein 1 (IAR1) genes, to save copper for the most essential functions and keep most IAA in conjugated forms, respectively, thereby protecting rice seedlings from As (III) damage [72].
The functional role of osa-miR528 in As (III) tolerance was directly demonstrated by overexpressing osa-miR528 (Ubi::MIR528) in a japonica rice Nipponbare cultivar, which is more tolerant to As(III) than Minghui 86 [21]. Compared with WT plants, Ubi::MIR528 plants presented highly upregulated expression of osa-miR528 in both the roots and leaves and showed As (III) sensitivity under stress conditions, which was likely due to the strong alteration in antioxidant enzyme activity and amino acid profiles and the impairment of the As (III) uptake, translocation, and tolerance systems of rice [21]. It is of interest to note that osa-miR528 exhibited opposite expression patterns in response to As (III) and As (V) exposure; specifically, osa-miR528 was upregulated upon As (III) stress but downregulated upon As (V) stress [73].

4.6. Heavy-Metal Stress

In addition to As stress, heavy metals represent a class of looming soil contaminants and environmental pollutants, and aluminum (Al) and cadmium (Cd), in particular, are extremely toxic to plants [74,75]. There is a growing body of evidence that miR528 plays a crucial regulatory role in the processes of heavy-metal stress response and/or tolerance. In rice, osa-miR528, which putatively targets an F-box/LRR repeat gene, MORE AXILLARY GROWTH 2 (MAX2), was strongly downregulated in rice seedling roots under Al stress [76]. Microarray-based analysis showed that upon Cd exposure, osa-miR528 was upregulated in rice seedling roots, and metal-responsive element (MRE) was identified in the promoter region of osa-MIR528, which lends further credence to the potential involvement of osa-miR528 in regulating Cd stress tolerance and highlights miR528 as an area of interest for further investigation into heavy-metal tolerance in plants [77]. Beyond transcriptional changes, further studies have demonstrated that miR528 modulates ROS homeostasis, which is critical to mitigating heavy-metal-induced oxidative stress. Specifically, under Cd stress, rice miR528 represses the expression of its target gene unknown copper-binding-like protein 23 (UCL23), a small copper protein involved in ROS accumulation, thereby reducing oxidative stress; meanwhile, miR528 is negatively regulated by the transcription factor WRKY transcription factor 51 (WRKY51), forming a WRKY51–miR528–UCL23 regulatory module that coordinates the rice response to Cd toxicity [78]. Similarly, Under chromium stress, the downregulation of zma-miR528 alleviates its repression on the SOD gene, thereby enhancing ROS scavenging capacity, mitigating oxidative damage, and improving root vitality and chromium stress tolerance, highlighting miR528 as a pivotal regulator in the antioxidant defense mechanism [79]. Furthermore, miR528 may mediate spatially distinct growth responses under heavy-metal stress. In maize, in response to Al stress, the accumulation of zma-miR528 was induced in primary roots but reduced in seminal roots, suggesting a role of zma-miR528 in mediating the differential growth responses to Al stress [23].

4.7. Nitrogen Homeostasis

There is growing recognition that miR528 plays an important role in the plant response to deficiencies in plant nutrients, such as nitrogen (N) and zinc (Zn), by modulating nutrient uptake, phloem-mediated long-distance transport, and nutrient homeostasis. It has been suggested that miR528 may act as a systemic signal in coordinating plants’ responsive physiological activities to alleviate nutrient deficiency stresses or toxicities [24]. In creeping bentgrass, miR528 was significantly suppressed under N deprivation, and overexpressing rice pri-miR528 in creeping bentgrass plants resulted in a significant enhancement in tolerance to N deficiency stress, which was associated with increases in biomass, total N accumulation and chlorophyll synthesis, reduction in AAO activity, and increases in NITRITE REDUCTASE (NiR) activity [16]. On the other hand, applying excessive quantities of chemical N fertilizer, coupled with a decrease in N use efficiency, resulted in both looming environmental problems and yield losses due to lodging under N-luxury conditions [80]. In maize, zma-miR528 was found to affect lignin content and composition and ameliorate lodging resistance in maize seedlings under N-luxury conditions by negatively regulating the expression of the ZmLACCASE3 (ZmLAC3) and ZmLACCASE5 (ZmLAC5) genes [81]. Building on this, recent studies have further revealed that under varying nitrogen conditions, zma-miR528 modulates Casparian strip formation by targeting the lignin-associated gene ZmLAC3, thereby influencing root apoplastic barrier development and ion homeostasis, and uncovering a novel regulatory link between nitrogen signaling and nutrient transport in maize [82].

5. Conclusions and Perspectives

Overall, as a monocot-specific miRNA in plants, miR528 has drawn considerable research interest in recent years because of its involvement in plant growth and development and in response to numerous biotic and abiotic stress conditions. There is growing recognition that miR528 acts in concert with many other miRNAs that have been found to be co-responsive to certain stress factors or targeting the same genes. As comprehensively summarized in Table 2, miR528 regulates diverse agronomic traits across major monocot crops, highlighting its potential as a key target for crop improvement strategies. However, the formation and evolution of the potentially monocot-specific regulatory networks in which miR528 participates are intriguing but remain elusive and deserve further investigation. A comprehensive understanding on the miR528-mediated gene regulatory networks in plant growth, development, and stress response cascades will facilitate the development of new strategies aimed at improving crop resilience and production.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms26157334/s1.

Author Contributions

Conceptualization, H.F.; formal analysis, H.F. and L.Z.; data curation, H.F., Y.H., Z.L., Z.W., and F.S.; writing—original draft preparation, H.F.; writing—review and editing, W.W.; supervision, W.W. All authors have read and agreed to the published version of the manuscript.

Funding

This research study was funded by Shandong Provincial Natural Science Foundation, grant number ZR2020QC118; China Postdoctoral Science Foundation, grant number 2020M672097; and the earmarked fund for CARS (grant No. CARS-15-34).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

No new data were created in this study. Data sharing is not applicable to this article.

Conflicts of Interest

The authors declare no conflicts of interest.

References

  1. Bartel, D.P. MicroRNAs: Genomics, Biogenesis, Mechanism, and Function. Cell 2004, 116, 281–297. [Google Scholar] [CrossRef]
  2. Yu, Y.; Jia, T.; Chen, X. The “how” and “Where” of Plant microRNAs. New Phytol. 2017, 216, 1002–1017. [Google Scholar] [CrossRef]
  3. Shahid, S.; Kim, G.; Johnson, N.R.; Wafula, E.; Wang, F.; Coruh, C.; Bernal-Galeano, V.; Phifer, T.; dePamphilis, C.W.; Westwood, J.H.; et al. MicroRNAs from the Parasitic Plant Cuscuta campestris Target Host Messenger RNAs. Nature 2018, 553, 82–85. [Google Scholar] [CrossRef]
  4. Wang, W.; Liu, D.; Zhang, X.; Chen, D.; Cheng, Y.; Shen, F. Plant MicroRNAs in Cross-Kingdom Regulation of Gene Expression. Int. J. Mol. Sci. 2018, 19, 2007. [Google Scholar] [CrossRef]
  5. Huang, C.-Y.; Wang, H.; Hu, P.; Hamby, R.; Jin, H. Small RNAs—Big Players in Plant-Microbe Interactions. Cell Host Microbe 2019, 26, 173–182. [Google Scholar] [CrossRef] [PubMed]
  6. Jones-Rhoades, M.W.; Bartel, D.P. Computational Identification of Plant microRNAs and Their Targets, Including a Stress-Induced miRNA. Mol. Cell 2004, 14, 787–799. [Google Scholar] [CrossRef]
  7. Song, X.; Li, Y.; Cao, X.; Qi, Y. MicroRNAs and Their Regulatory Roles in Plant-Environment Interactions. Annu. Rev. Plant Biol. 2019, 70, 489–525. [Google Scholar] [CrossRef] [PubMed]
  8. Zhao, X.Y.; Hong, P.; Wu, J.Y.; Chen, X.B.; Ye, X.G.; Pan, Y.Y.; Wang, J.; Zhang, X.S. The Tae-miR408-Mediated Control of TaTOC1 Genes Transcription Is Required for the Regulation of Heading Time in Wheat. Plant Physiol. 2016, 170, 1578–1594. [Google Scholar] [CrossRef]
  9. Bai, Q.; Wang, X.; Chen, X.; Shi, G.; Liu, Z.; Guo, C.; Xiao, K. Wheat miRNA TaemiR408 Acts as an Essential Mediator in Plant Tolerance to Pi Deprivation and Salt Stress via Modulating Stress-Associated Physiological Processes. Front. Plant Sci. 2018, 9, 499. [Google Scholar] [CrossRef] [PubMed]
  10. Zhang, H.; Zhang, J.; Yan, J.; Gou, F.; Mao, Y.; Tang, G.; Botella, J.R.; Zhu, J.-K. Short Tandem Target Mimic Rice Lines Uncover Functions of miRNAs in Regulating Important Agronomic Traits. Proc. Natl. Acad. Sci. USA 2017, 114, 5277–5282. [Google Scholar] [CrossRef]
  11. Li, Y.; Cao, X.-L.; Zhu, Y.; Yang, X.-M.; Zhang, K.-N.; Xiao, Z.-Y.; Wang, H.; Zhao, J.-H.; Zhang, L.-L.; Li, G.-B.; et al. Osa-miR398b Boosts H2 O2 Production and Rice Blast Disease-Resistance via Multiple Superoxide Dismutases. New Phytol. 2019, 222, 1507–1522. [Google Scholar] [CrossRef]
  12. Li, Y.; Zhang, Q.; Zhang, J.; Wu, L.; Qi, Y.; Zhou, J.-M. Identification of microRNAs Involved in Pathogen-Associated Molecular Pattern-Triggered Plant Innate Immunity. Plant Physiol. 2010, 152, 2222–2231. [Google Scholar] [CrossRef] [PubMed]
  13. Xu, W.; Meng, Y.; Wise, R.P. Mla- and Rom1-Mediated Control of microRNA398 and Chloroplast Copper/Zinc Superoxide Dismutase Regulates Cell Death in Response to the Barley Powdery Mildew Fungus. New Phytol. 2014, 201, 1396–1412. [Google Scholar] [CrossRef] [PubMed]
  14. Yang, R.; Li, P.; Mei, H.; Wang, D.; Sun, J.; Yang, C.; Hao, L.; Cao, S.; Chu, C.; Hu, S.; et al. Fine-Tuning of MiR528 Accumulation Modulates Flowering Time in Rice. Mol. Plant 2019, 12, 1103–1113. [Google Scholar] [CrossRef]
  15. Zhang, Y.-C.; He, R.-R.; Lian, J.-P.; Zhou, Y.-F.; Zhang, F.; Li, Q.-F.; Yu, Y.; Feng, Y.-Z.; Yang, Y.-W.; Lei, M.-Q.; et al. OsmiR528 Regulates Rice-Pollen Intine Formation by Targeting an Uclacyanin to Influence Flavonoid Metabolism. Proc. Natl. Acad. Sci. USA 2020, 117, 727–732. [Google Scholar] [CrossRef]
  16. Yuan, S.; Li, Z.; Li, D.; Yuan, N.; Hu, Q.; Luo, H. Constitutive Expression of Rice MicroRNA528 Alters Plant Development and Enhances Tolerance to Salinity Stress and Nitrogen Starvation in Creeping Bentgrass. Plant Physiol. 2015, 169, 576–593. [Google Scholar] [CrossRef]
  17. Jin, X.; Fu, Z.; Lv, P.; Peng, Q.; Ding, D.; Li, W.; Tang, J. Identification and Characterization of microRNAs during Maize Grain Filling. PLoS ONE 2015, 10, e0125800. [Google Scholar] [CrossRef]
  18. Wu, J.; Yang, R.; Yang, Z.; Yao, S.; Zhao, S.; Wang, Y.; Li, P.; Song, X.; Jin, L.; Zhou, T.; et al. ROS Accumulation and Antiviral Defence Control by microRNA528 in Rice. Nat. Plants 2017, 3, 16203. [Google Scholar] [CrossRef] [PubMed]
  19. Zhu, H.; Zhang, Y.; Tang, R.; Qu, H.; Duan, X.; Jiang, Y. Banana sRNAome and Degradome Identify microRNAs Functioning in Differential Responses to Temperature Stress. BMC Genom. 2019, 20, 33. [Google Scholar] [CrossRef]
  20. Kantar, M.; Lucas, S.J.; Budak, H. miRNA Expression Patterns of Triticum Dicoccoides in Response to Shock Drought Stress. Planta 2011, 233, 471–484. [Google Scholar] [CrossRef]
  21. Liu, Q.; Hu, H.; Zhu, L.; Li, R.; Feng, Y.; Zhang, L.; Yang, Y.; Liu, X.; Zhang, H. Involvement of miR528 in the Regulation of Arsenite Tolerance in Rice (Oryza sativa L.). J. Agric. Food Chem. 2015, 63, 8849–8861. [Google Scholar] [CrossRef]
  22. Li, T.; Li, H.; Zhang, Y.-X.; Liu, J.-Y. Identification and Analysis of Seven H2O2-Responsive miRNAs and 32 New miRNAs in the Seedlings of Rice (Oryza sativa L. Ssp. Indica). Nucleic Acids Res. 2011, 39, 2821–2833. [Google Scholar] [CrossRef]
  23. Kong, X.; Zhang, M.; Xu, X.; Li, X.; Li, C.; Ding, Z. System Analysis of microRNAs in the Development and Aluminium Stress Responses of the Maize Root System. Plant Biotechnol. J. 2014, 12, 1108–1121. [Google Scholar] [CrossRef]
  24. Zeng, H.; Wang, G.; Hu, X.; Wang, H.; Du, L.; Zhu, Y. Role of microRNAs in Plant Responses to Nutrient Stress. Plant Soil 2014, 374, 1005–1021. [Google Scholar] [CrossRef]
  25. Liu, B.; Li, P.; Li, X.; Liu, C.; Cao, S.; Chu, C.; Cao, X. Loss of Function of OsDCL1 Affects microRNA Accumulation and Causes Developmental Defects in Rice. Plant Physiol. 2005, 139, 296–305. [Google Scholar] [CrossRef] [PubMed]
  26. Kurtoglu, K.Y.; Kantar, M.; Budak, H. New Wheat microRNA Using Whole-Genome Sequence. Funct. Integr. Genom. 2014, 14, 363–379. [Google Scholar] [CrossRef]
  27. Zhang, L.; Chia, J.-M.; Kumari, S.; Stein, J.C.; Liu, Z.; Narechania, A.; Maher, C.A.; Guill, K.; McMullen, M.D.; Ware, D. A Genome-Wide Characterization of microRNA Genes in Maize. PLOS Genet. 2009, 5, e1000716. [Google Scholar] [CrossRef]
  28. Paterson, A.H.; Bowers, J.E.; Bruggmann, R.; Dubchak, I.; Grimwood, J.; Gundlach, H.; Haberer, G.; Hellsten, U.; Mitros, T.; Poliakov, A.; et al. The Sorghum Bicolor Genome and the Diversification of Grasses. Nature 2009, 457, 551–556. [Google Scholar] [CrossRef]
  29. Zanca, A.S.; Vicentini, R.; Ortiz-Morea, F.A.; Del Bem, L.E.V.; da Silva, M.J.; Vincentz, M.; Nogueira, F.T.S. Identification and Expression Analysis of microRNAs and Targets in the Biofuel Crop Sugarcane. BMC Plant Biol. 2010, 10, 260. [Google Scholar] [CrossRef] [PubMed]
  30. Guo, Z.; Kuang, Z.; Zhao, Y.; Deng, Y.; He, H.; Wan, M.; Tao, Y.; Wang, D.; Wei, J.; Li, L.; et al. PmiREN2.0: From Data Annotation to Functional Exploration of Plant microRNAs. Nucleic Acids Res. 2022, 50, D1475–D1482. [Google Scholar] [CrossRef] [PubMed]
  31. Tamura, K.; Stecher, G.; Peterson, D.; Filipski, A.; Kumar, S. MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0. Mol. Biol. Evol. 2013, 30, 2725–2729. [Google Scholar] [CrossRef] [PubMed]
  32. Van Bel, M.; Diels, T.; Vancaester, E.; Kreft, L.; Botzki, A.; Van de Peer, Y.; Coppens, F.; Vandepoele, K. PLAZA 4.0: An Integrative Resource for Functional, Evolutionary and Comparative Plant Genomics. Nucleic Acids Res. 2018, 46, D1190–D1196. [Google Scholar] [CrossRef]
  33. Soreng, R.J.; Peterson, P.M.; Romaschenko, K.; Davidse, G.; Zuloaga, F.O.; Judziewicz, E.J.; Filgueiras, T.S.; Davis, J.I.; Morrone, O. A Worldwide Phylogenetic Classification of the Poaceae (Gramineae). J. Syst. Evol. 2015, 53, 117–137. [Google Scholar] [CrossRef]
  34. Grass Phylogeny Working Group II New Grass Phylogeny Resolves Deep Evolutionary Relationships and Discovers C4 Origins. New Phytol. 2012, 193, 304–312. [CrossRef]
  35. Zhu, H.; Chen, C.; Zeng, J.; Yun, Z.; Liu, Y.; Qu, H.; Jiang, Y.; Duan, X.; Xia, R. MicroRNA528, a Hub Regulator Modulating ROS Homeostasis via Targeting of a Diverse Set of Genes Encoding Copper-Containing Proteins in Monocots. New Phytol. 2020, 225, 385–399. [Google Scholar] [CrossRef]
  36. Dai, X.; Zhuang, Z.; Zhao, P.X. psRNATarget: A Plant Small RNA Target Analysis Server (2017 Release). Nucleic Acids Res. 2018, 46, W49–W54. [Google Scholar] [CrossRef]
  37. Pilon, M. The Copper microRNAs. New Phytol. 2017, 213, 1030–1035. [Google Scholar] [CrossRef]
  38. Chen, C.; Wu, Y.; Li, J.; Wang, X.; Zeng, Z.; Xu, J.; Liu, Y.; Feng, J.; Chen, H.; He, Y.; et al. TBtools-II: A “One for All, All for One” Bioinformatics Platform for Biological Big-Data Mining. Mol. Plant 2023, 16, 1733–1742. [Google Scholar] [CrossRef]
  39. Li, X.-K.; Qin, X.-M.; Cui, J.-L. Transcriptional Regulatory Mechanisms of Flavonoid Metabolism from Flower Bud to Flower in Hemerocallis Citrina. Sci. Hortic. 2024, 326, 112766. [Google Scholar] [CrossRef]
  40. Li, T.; Tang, S.; Li, W.; Zhang, S.; Wang, J.; Pan, D.; Lin, Z.; Ma, X.; Chang, Y.; Liu, B.; et al. Genome Evolution and Initial Breeding of the Triticeae Grass Leymus Chinensis Dominating the Eurasian Steppe. Proc. Natl. Acad. Sci. USA 2023, 120, e2308984120. [Google Scholar] [CrossRef] [PubMed]
  41. Zhao, J.; Liu, X.; Wang, M.; Xie, L.; Wu, Z.; Yu, J.; Wang, Y.; Zhang, Z.; Jia, Y.; Liu, Q. The miR528-D3 Module Regulates Plant Height in Rice by Modulating the Gibberellin and Abscisic Acid Metabolisms. Rice 2022, 15, 27. [Google Scholar] [CrossRef]
  42. Luján-Soto, E.; Aguirre de la Cruz, P.I.; Juárez-González, V.T.; Reyes, J.L.; Sanchez, M.d.l.P.; Dinkova, T.D. Transcriptional Regulation of Zma-MIR528a by Action of Nitrate and Auxin in Maize. Int. J. Mol. Sci. 2022, 23, 15718. [Google Scholar] [CrossRef] [PubMed]
  43. Li, D.; Liu, Z.; Gao, L.; Wang, L.; Gao, M.; Jiao, Z.; Qiao, H.; Yang, J.; Chen, M.; Yao, L.; et al. Genome-Wide Identification and Characterization of microRNAs in Developing Grains of Zea mays L. PLoS ONE 2016, 11, e0153168. [Google Scholar] [CrossRef]
  44. Kang, M.; Zhao, Q.; Zhu, D.; Yu, J. Characterization of microRNAs Expression during Maize Seed Development. BMC Genom. 2012, 13, 360. [Google Scholar] [CrossRef] [PubMed]
  45. Zhu, Q.-H.; Spriggs, A.; Matthew, L.; Fan, L.; Kennedy, G.; Gubler, F.; Helliwell, C. A Diverse Set of microRNAs and microRNA-like Small RNAs in Developing Rice Grains. Genome Res. 2008, 18, 1456–1465. [Google Scholar] [CrossRef]
  46. Xue, L.-J.; Zhang, J.-J.; Xue, H.-W. Characterization and Expression Profiles of miRNAs in Rice Seeds. Nucleic Acids Res. 2009, 37, 916–930. [Google Scholar] [CrossRef]
  47. Luo, Y.-C.; Zhou, H.; Li, Y.; Chen, J.-Y.; Yang, J.-H.; Chen, Y.-Q.; Qu, L.-H. Rice Embryogenic Calli Express a Unique Set of microRNAs, Suggesting Regulatory Roles of microRNAs in Plant Post-Embryogenic Development. FEBS Lett. 2006, 580, 5111–5116. [Google Scholar] [CrossRef]
  48. Alejandri-Ramírez, N.D.; Chávez-Hernández, E.C.; Contreras-Guerra, J.L.; Reyes, J.L.; Dinkova, T.D. Small RNA Differential Expression and Regulation in Tuxpeño Maize Embryogenic Callus Induction and Establishment. Plant Physiol. Biochem. PPB 2018, 122, 78–89. [Google Scholar] [CrossRef]
  49. Chávez-Hernández, E.C.; Alejandri-Ramírez, N.D.; Juárez-González, V.T.; Dinkova, T.D. Maize miRNA and Target Regulation in Response to Hormone Depletion and Light Exposure during Somatic Embryogenesis. Front. Plant Sci. 2015, 6, 555. [Google Scholar] [CrossRef]
  50. Chen, C.-J.; Liu, Q.; Zhang, Y.-C.; Qu, L.-H.; Chen, Y.-Q.; Gautheret, D. Genome-Wide Discovery and Analysis of microRNAs and Other Small RNAs from Rice Embryogenic Callus. RNA Biol. 2011, 8, 538–547. [Google Scholar] [CrossRef] [PubMed]
  51. Luján-Soto, E.; Juárez-González, V.T.; Reyes, J.L.; Dinkova, T.D. MicroRNA Zma-miR528 Versatile Regulation on Target mRNAs during Maize Somatic Embryogenesis. Int. J. Mol. Sci. 2021, 22, 5310. [Google Scholar] [CrossRef]
  52. Shen, Y.; Jiang, Z.; Lu, S.; Lin, H.; Gao, S.; Peng, H.; Yuan, G.; Liu, L.; Zhang, Z.; Zhao, M.; et al. Combined Small RNA and Degradome Sequencing Reveals microRNA Regulation during Immature Maize Embryo Dedifferentiation. Biochem. Biophys. Res. Commun. 2013, 441, 425–430. [Google Scholar] [CrossRef]
  53. Wu, J.; Yang, Z.; Wang, Y.; Zheng, L.; Ye, R.; Ji, Y.; Zhao, S.; Ji, S.; Liu, R.; Xu, L.; et al. Viral-Inducible Argonaute18 Confers Broad-Spectrum Virus Resistance in Rice by Sequestering a Host microRNA. Elife 2015, 4, e05733. [Google Scholar] [CrossRef] [PubMed]
  54. Yao, S.; Yang, Z.; Yang, R.; Huang, Y.; Guo, G.; Kong, X.; Lan, Y.; Zhou, T.; Wang, H.; Wang, W.; et al. Transcriptional Regulation of miR528 by OsSPL9 Orchestrates Antiviral Response in Rice. Mol. Plant 2019, 12, 1114–1122. [Google Scholar] [CrossRef] [PubMed]
  55. Yao, S.; Kang, J.; Guo, G.; Yang, Z.; Huang, Y.; Lan, Y.; Zhou, T.; Wang, L.; Wei, C.; Xu, Z.; et al. The Key Micronutrient Copper Orchestrates Broad-Spectrum Virus Resistance in Rice. Sci. Adv. 2022, 8, eabm0660. [Google Scholar] [CrossRef]
  56. Liu, K.; Ma, X.; Zhao, L.; Lai, X.; Chen, J.; Lang, X.; Han, Q.; Wan, X.; Li, C. Comprehensive Transcriptomic Analysis of Three Varieties with Different Brown Planthopper-Resistance Identifies Leaf Sheath lncRNAs in Rice. BMC Plant Biol. 2023, 23, 367. [Google Scholar] [CrossRef] [PubMed]
  57. Qu, Q.; Liu, N.; Su, Q.; Liu, X.; Jia, H.; Liu, Y.; Sun, M.; Cao, Z.; Dong, J. MicroRNAs Involved in the Trans-Kingdom Gene Regulation in the Interaction of Maize Kernels and Fusarium Verticillioides. Int. J. Biol. Macromol. 2023, 242, 125046. [Google Scholar] [CrossRef]
  58. Ding, D.; Zhang, L.; Wang, H.; Liu, Z.; Zhang, Z.; Zheng, Y. Differential Expression of miRNAs in Response to Salt Stress in Maize Roots. Ann. Bot. 2009, 103, 29–38. [Google Scholar] [CrossRef]
  59. Wang, M.; Guo, W.; Li, J.; Pan, X.; Pan, L.; Zhao, J.; Zhang, Y.; Cai, S.; Huang, X.; Wang, A.; et al. The miR528-AO Module Confers Enhanced Salt Tolerance in Rice by Modulating the Ascorbic Acid and Abscisic Acid Metabolism and ROS Scavenging. J. Agric. Food Chem. 2021, 69, 8634–8648. [Google Scholar] [CrossRef]
  60. Hivrale, V.; Zheng, Y.; Puli, C.O.R.; Jagadeeswaran, G.; Gowdu, K.; Kakani, V.G.; Barakat, A.; Sunkar, R. Characterization of Drought- and Heat-Responsive microRNAs in Switchgrass. Plant Sci. Int. J. Exp. Plant Biol. 2016, 242, 214–223. [Google Scholar] [CrossRef]
  61. Wang, Z.; Pu, H.; Shan, S.; Zhang, P.; Li, J.; Song, H.; Xu, X. Melatonin Enhanced Chilling Tolerance and Alleviated Peel Browning of Banana Fruit under Low Temperature Storage. Postharvest Biol. Technol. 2021, 179, 111571. [Google Scholar] [CrossRef]
  62. Hong, Z.; Xu, H.; Shen, Y.; Liu, C.; Guo, F.; Muhammad, S.; Zhang, Y.; Niu, H.; Li, S.; Zhou, W.; et al. Bioengineering for Robust Tolerance against Cold and Drought Stresses via Co-Overexpressing Three Cu-miRNAs in Major Food Crops. Cell Rep. 2024, 43, 114828. [Google Scholar] [CrossRef]
  63. Wang, H.; Jia, Y.; Bai, X.; Wang, J.; Liu, G.; Wang, H.; Wu, Y.; Xin, J.; Ma, H.; Liu, Z.; et al. Whole-Transcriptome Profiling and Identification of Cold Tolerance-Related ceRNA Networks in Japonica Rice Varieties. Front. Plant Sci. 2024, 15, 1260591. [Google Scholar] [CrossRef]
  64. Ferreira, T.H.; Gentile, A.; Vilela, R.D.; Costa, G.G.L.; Dias, L.I.; Endres, L.; Menossi, M. microRNAs Associated with Drought Response in the Bioenergy Crop Sugarcane (Saccharum spp.). PLoS ONE 2012, 7, e46703. [Google Scholar] [CrossRef]
  65. Liu, Z.; Kumari, S.; Zhang, L.; Zheng, Y.; Ware, D. Characterization of miRNAs in Response to Short-Term Waterlogging in Three Inbred Lines of Zea mays. PLoS ONE 2012, 7, e39786. [Google Scholar] [CrossRef]
  66. Chen, J.; Zhong, Y.; Qi, X. LncRNA TCONS_00021861 Is Functionally Associated with Drought Tolerance in Rice (Oryza sativa L.) via Competing Endogenous RNA Regulation. BMC Plant Biol. 2021, 21, 410. [Google Scholar] [CrossRef]
  67. Fan, B.; Sun, F.; Yu, Z.; Zhang, X.; Yu, X.; Wu, J.; Yan, X.; Zhao, Y.; Nie, L.; Fang, Y.; et al. Integrated Analysis of Small RNAs, Transcriptome and Degradome Sequencing Reveal the Drought Stress Network in Agropyron Mongolicum Keng. Front. Plant Sci. 2022, 13, 976684. [Google Scholar] [CrossRef]
  68. Kumar, D.; Ramkumar, M.K.; Dutta, B.; Kumar, A.; Pandey, R.; Jain, P.K.; Gaikwad, K.; Mishra, D.C.; Chaturvedi, K.K.; Rai, A.; et al. Integration of miRNA Dynamics and Drought Tolerant QTLs in Rice Reveals the Role of miR2919 in Drought Stress Response. BMC Genom. 2023, 24, 526. [Google Scholar] [CrossRef] [PubMed]
  69. Ghorbanzadeh, Z.; Hamid, R.; Jacob, F.; Mirzaei, M.; Zeinalabedini, M.; Abdirad, S.; Atwell, B.J.; Haynes, P.A.; Ghaffari, M.R.; Salekdeh, G.H. MicroRNA Profiling of Root Meristematic Zone in Contrasting Genotypes Reveals Novel Insight into in Rice Response to Water Deficiency. J. Plant Growth Regul. 2023, 42, 3814–3834. [Google Scholar] [CrossRef]
  70. Ali, W.; Isayenkov, S.V.; Zhao, F.-J.; Maathuis, F.J.M. Arsenite Transport in Plants. Cell. Mol. Life Sci. CMLS 2009, 66, 2329–2339. [Google Scholar] [CrossRef] [PubMed]
  71. Meharg, A.A. Arsenic in Rice—Understanding a New Disaster for South-East Asia. Trends Plant Sci. 2004, 9, 415–417. [Google Scholar] [CrossRef]
  72. Liu, Q.; Zhang, H. Molecular Identification and Analysis of Arsenite Stress-Responsive miRNAs in Rice. J. Agric. Food Chem. 2012, 60, 6524–6536. [Google Scholar] [CrossRef] [PubMed]
  73. Sharma, D.; Tiwari, M.; Lakhwani, D.; Tripathi, R.D.; Trivedi, P.K. Differential Expression of microRNAs by Arsenate and Arsenite Stress in Natural Accessions of Rice. Met. Integr. Biometal Sci. 2015, 7, 174–187. [Google Scholar] [CrossRef]
  74. Kikui, S.; Sasaki, T.; Maekawa, M.; Miyao, A.; Hirochika, H.; Matsumoto, H.; Yamamoto, Y. Physiological and Genetic Analyses of Aluminium Tolerance in Rice, Focusing on Root Growth during Germination. J. Inorg. Biochem. 2005, 99, 1837–1844. [Google Scholar] [CrossRef]
  75. Zhang, M.; Lu, X.; Li, C.; Zhang, B.; Zhang, C.; Zhang, X.-S.; Ding, Z. Auxin Efflux Carrier ZmPGP1 Mediates Root Growth Inhibition under Aluminum Stress. Plant Physiol. 2018, 177, 819–832. [Google Scholar] [CrossRef]
  76. Lima, J.C.; Arenhart, R.A.; Margis-Pinheiro, M.; Margis, R. Aluminum Triggers Broad Changes in microRNA Expression in Rice Roots. Genet. Mol. Res. GMR 2011, 10, 2817–2832. [Google Scholar] [CrossRef]
  77. Ding, Y.; Chen, Z.; Zhu, C. Microarray-Based Analysis of Cadmium-Responsive microRNAs in Rice (Oryza sativa). J. Exp. Bot. 2011, 62, 3563–3573. [Google Scholar] [CrossRef]
  78. Tan, J.; Zhang, L.; Liu, C.; Hong, Z.; Wu, X.; Zhang, Y.; Fahad, M.; Shen, Y.; Bian, J.; He, H.; et al. UCL23 Hierarchically Regulated by WRKY51-miR528 Mediates Cadmium Uptake, Tolerance, and Accumulation in Rice. Cell Rep. 2025, 44, 115336. [Google Scholar] [CrossRef]
  79. Adhikari, A.; Roy, D.; Adhikari, S.; Saha, S.; Ghosh, P.K.; Shaw, A.K.; Hossain, Z. microRNAomic Profiling of Maize Root Reveals Multifaceted Mechanisms to Cope with Cr (VI) Stress. Plant Physiol. Biochem. PPB 2023, 198, 107693. [Google Scholar] [CrossRef] [PubMed]
  80. Khan, S.; Anwar, S.; Kuai, J.; Noman, A.; Shahid, M.; Din, M.; Ali, A.; Zhou, G. Alteration in Yield and Oil Quality Traits of Winter Rapeseed by Lodging at Different Planting Density and Nitrogen Rates. Sci. Rep. 2018, 8, 634. [Google Scholar] [CrossRef]
  81. Sun, Q.; Liu, X.; Yang, J.; Liu, W.; Du, Q.; Wang, H.; Fu, C.; Li, W.-X. MicroRNA528 Affects Lodging Resistance of Maize by Regulating Lignin Biosynthesis under Nitrogen-Luxury Conditions. Mol. Plant 2018, 11, 806–814. [Google Scholar] [CrossRef] [PubMed]
  82. Guo, Y.; Wang, Y.; Chen, H.; Du, Q.; Wang, Z.; Gong, X.; Sun, Q.; Li, W.-X. Nitrogen Supply Affects Ion Homeostasis by Modifying Root Casparian Strip Formation through the miR528-LAC3 Module in Maize. Plant Commun. 2023, 4, 100553. [Google Scholar] [CrossRef] [PubMed]
Ijms 26 07334 g001
Figure 1. The sequence alignment and phylogenetic analysis of miR528. (A) Alignment of mature miR528 sequences. miR528 derived from 17 monocot plant species, including Asparagus officinalis L. (aof), Brachypodium distachyon (L.) P.Beauv. (bdi), Musa acuminata Colla (mac), Oryza glaberrima Steud. (ogl), Oryza nivara S.D.Sharma and Shastry (oni), Oryza rufipogon Griff. (Oru), Oryza sativa L. (osa), Panicum hallii Vasey (pha), Panicum virgatum L. (pvi), Phalaenopsis aphrodite Rchb.f. (pap), Phoenix dactylifera L. (pda), Saccharum hybrid cultivar (scu), Setaria italica (L.) P. Beauvois (sit), Sorghum bicolor (L.) Moench (sbi), Spirodela polyrhiza (L.) Schleid. (spo), Triticum aestivum L. (tae), and Zea mays L. (zma). The black-shaded blocks indicate highly conserved residues. (B) The phylogenetic tree reconstructed with precursors of miR528. We aligned the miR528 precursor sequences of all the 17 monocot plant species using Molecular Evolutionary Genetics Analysis (MEGA) version 6.06 software [31] with MUltiple Sequence Comparison by Log-Expectation (MUSCLE) and reconstructed the phylogenetic tree using the maximum likelihood (ML) method using the general time reversible (GTR) substitution model with the default set of gamma distribution among-site rate variation. To compare the similarities and differences of the ML phylogenetic tree, we also reconstructed the phylogenetic tree with the maximum parsimony (MP) method using MEGA (Supplementary Figure S1). The phylogenetic tree of the 17 monocot plants used in the study was gathered from Monocots Plant Annotated Genomes Database (PLAZA) 4.5 (https://bioinformatics.psb.ugent.be/plaza/, accessed on 6 December 2024) [32]. The 17 monocot plants belong to five orders of the Liliopsida. The five orders are as follows: Alismatales, Asparagales, Principes, Zingiberales, and Poales. Poaceae is a large and nearly ubiquitous family of monocotyledonous flowering plants known as grasses, containing two sister lineages (or clades): the Panicoideae, Arundinoideae, Chloridoideae, Micrairoideae, Aristidoideae, and Danthonioideae (PACMAD) clade and the Bambusoideae, Oryzoideae, and Pooideae (BOP) clade. The name of the PACMAD clade comes from the first initials of the six included subfamilies, i.e., Panicoideae, Arundinoideae, Chloridoideae, Micrairoideae, Aristidoideae, and Danthonioideae [33]. The BOP clade contains three subfamilies from whose initials its name derives: bamboos (Bambusoideae); Oryzoideae, including rice; and Pooideae, including important cereal crops such as wheat [34].
Figure 1. The sequence alignment and phylogenetic analysis of miR528. (A) Alignment of mature miR528 sequences. miR528 derived from 17 monocot plant species, including Asparagus officinalis L. (aof), Brachypodium distachyon (L.) P.Beauv. (bdi), Musa acuminata Colla (mac), Oryza glaberrima Steud. (ogl), Oryza nivara S.D.Sharma and Shastry (oni), Oryza rufipogon Griff. (Oru), Oryza sativa L. (osa), Panicum hallii Vasey (pha), Panicum virgatum L. (pvi), Phalaenopsis aphrodite Rchb.f. (pap), Phoenix dactylifera L. (pda), Saccharum hybrid cultivar (scu), Setaria italica (L.) P. Beauvois (sit), Sorghum bicolor (L.) Moench (sbi), Spirodela polyrhiza (L.) Schleid. (spo), Triticum aestivum L. (tae), and Zea mays L. (zma). The black-shaded blocks indicate highly conserved residues. (B) The phylogenetic tree reconstructed with precursors of miR528. We aligned the miR528 precursor sequences of all the 17 monocot plant species using Molecular Evolutionary Genetics Analysis (MEGA) version 6.06 software [31] with MUltiple Sequence Comparison by Log-Expectation (MUSCLE) and reconstructed the phylogenetic tree using the maximum likelihood (ML) method using the general time reversible (GTR) substitution model with the default set of gamma distribution among-site rate variation. To compare the similarities and differences of the ML phylogenetic tree, we also reconstructed the phylogenetic tree with the maximum parsimony (MP) method using MEGA (Supplementary Figure S1). The phylogenetic tree of the 17 monocot plants used in the study was gathered from Monocots Plant Annotated Genomes Database (PLAZA) 4.5 (https://bioinformatics.psb.ugent.be/plaza/, accessed on 6 December 2024) [32]. The 17 monocot plants belong to five orders of the Liliopsida. The five orders are as follows: Alismatales, Asparagales, Principes, Zingiberales, and Poales. Poaceae is a large and nearly ubiquitous family of monocotyledonous flowering plants known as grasses, containing two sister lineages (or clades): the Panicoideae, Arundinoideae, Chloridoideae, Micrairoideae, Aristidoideae, and Danthonioideae (PACMAD) clade and the Bambusoideae, Oryzoideae, and Pooideae (BOP) clade. The name of the PACMAD clade comes from the first initials of the six included subfamilies, i.e., Panicoideae, Arundinoideae, Chloridoideae, Micrairoideae, Aristidoideae, and Danthonioideae [33]. The BOP clade contains three subfamilies from whose initials its name derives: bamboos (Bambusoideae); Oryzoideae, including rice; and Pooideae, including important cereal crops such as wheat [34].
Ijms 26 07334 g001
Ijms 26 07334 g002
Figure 2. The target genes of miR528. (A) The sequence logos of miR528 target sites. Toolkit for Biologists integrating various biological data handling tools (TBtools) was used to draw the sequence logos (https://github.com/CJ-Chen/TBtools/, accessed on 25 December 2024) [38]. The sequence logos of miR528 target sites are conserved in rice, corn, wheat, millet, and switchgrass. These conserved target genes encode three types proteins, including plastocyanin-like domain-containing proteins, F-box domain- and (or) leucine-rich repeat (LRR)-containing proteins, and the copper/zinc superoxide dismutase. osa-miR528, zma-miR528, tae-miR528, sit-miR528, and pvi-miR528 represent the sequence logos of all the predicted miR528 target sites in rice, corn, wheat, millet, and switchgrass, respectively. (B–D) The conserved target genes of miR528. The target genes of miR528 were predicted by searching the transcripts for complementary sequences using the psRNATarget server with a more stringent cut-off threshold: maximum expectation (E) = 3.0 (https://www.zhaolab.org/psRNATarget/, accessed on 26 December 2024) [36]. Degradome sequencing data were downloaded from the PmiREN database. The upper parts of B-D present the alignments of miR528 with its target sequences, and the lower parts are the frequency of the 5′ end of the degradome tags within the full-length target transcripts. The solid lines indicate matched RNA base pairs. One dot shows G-U mismatch, and two dots represent other types of mismatch. (B) LOC_Os07g38290 and GRMZM2G107562, encoding a plastocyanin-like domain-containing protein, are the targets of miR528 in rice and maize, respectively; (C) LOC_Os06g06050 and GRMZM2G040278, encoding F-box domain- and (or) LRR-containing proteins, are the targets of miR528 in rice and maize, respectively; (D) LOC_Os08g44770 and GRMZM2G106928, encoding a copper/zinc superoxide dismutase, are the targets of miR528 in rice and maize, respectively. (E–H) Examples of non-conserved target genes of miR528 predicted in different monocot species, including genes involved in flowering regulation, glycosylation, alkaloid biosynthesis, and transcriptional regulation. The black line represents intron, the rectangle filled yellow represents exon, and miRNA complementary sites (red) are shown. The RNA sequences of each complementary site from 5′ to 3′ and the miR528 sequence from 3′ to 5′ are shown in the expanded regions.
Figure 2. The target genes of miR528. (A) The sequence logos of miR528 target sites. Toolkit for Biologists integrating various biological data handling tools (TBtools) was used to draw the sequence logos (https://github.com/CJ-Chen/TBtools/, accessed on 25 December 2024) [38]. The sequence logos of miR528 target sites are conserved in rice, corn, wheat, millet, and switchgrass. These conserved target genes encode three types proteins, including plastocyanin-like domain-containing proteins, F-box domain- and (or) leucine-rich repeat (LRR)-containing proteins, and the copper/zinc superoxide dismutase. osa-miR528, zma-miR528, tae-miR528, sit-miR528, and pvi-miR528 represent the sequence logos of all the predicted miR528 target sites in rice, corn, wheat, millet, and switchgrass, respectively. (B–D) The conserved target genes of miR528. The target genes of miR528 were predicted by searching the transcripts for complementary sequences using the psRNATarget server with a more stringent cut-off threshold: maximum expectation (E) = 3.0 (https://www.zhaolab.org/psRNATarget/, accessed on 26 December 2024) [36]. Degradome sequencing data were downloaded from the PmiREN database. The upper parts of B-D present the alignments of miR528 with its target sequences, and the lower parts are the frequency of the 5′ end of the degradome tags within the full-length target transcripts. The solid lines indicate matched RNA base pairs. One dot shows G-U mismatch, and two dots represent other types of mismatch. (B) LOC_Os07g38290 and GRMZM2G107562, encoding a plastocyanin-like domain-containing protein, are the targets of miR528 in rice and maize, respectively; (C) LOC_Os06g06050 and GRMZM2G040278, encoding F-box domain- and (or) LRR-containing proteins, are the targets of miR528 in rice and maize, respectively; (D) LOC_Os08g44770 and GRMZM2G106928, encoding a copper/zinc superoxide dismutase, are the targets of miR528 in rice and maize, respectively. (E–H) Examples of non-conserved target genes of miR528 predicted in different monocot species, including genes involved in flowering regulation, glycosylation, alkaloid biosynthesis, and transcriptional regulation. The black line represents intron, the rectangle filled yellow represents exon, and miRNA complementary sites (red) are shown. The RNA sequences of each complementary site from 5′ to 3′ and the miR528 sequence from 3′ to 5′ are shown in the expanded regions.
Ijms 26 07334 g002
Table 1. The summary of MIR528 from 17 monocot plant species.
Table 1. The summary of MIR528 from 17 monocot plant species.
Locus IDOrganismGenome Location aOverlapping Gene b
aof-MIR528Asparagus officinalis L.NC_033797.1:10579184~10579302,+×
bdi-MIR528Brachypodium distachyon (L.) P.Beauv.Bd1:73295140~73295266,−BRADI_1g76465v3
mac-MIR528Musa acuminata CollaChr8:10341398~10341507,−GSMUA_Achr8G13592_001
ogl-MIR528Oryza glaberrima Steud.GL455988.1:1549995~1550120,+ORGLA03G0021200
oni-MIR528Oryza nivara S.D.Sharma and ShastryChr3:1341691~1341816,+ONIVA03G01960
oru-MIR528Oryza rufipogon Griff.HG417167.1:1412714~1412839,+ORUFI03G02090
osa-MIR528Oryza sativa L.Chr3:1667310~1667435,+LOC_Os03g03724
pha-MIR528Panicum hallii VaseyChr09:69425755~69425882,−×
pvi-MIR528aPanicum virgatum L.Chr01N:69192314~69192445,+Pavir.Aa02527.1
pvi-MIR528bPanicum virgatum L.Chr09K:87418444~87418571,−×
pvi-MIR528cPanicum virgatum L.Chr09N:120070864~120070991,−×
pap-MIR528Phalaenopsis aphrodite Rchb.f.NEWO01000071.1:538724~538838,+×
pda-MIR528Phoenix dactylifera L.NW_008247750.1:18899~19050,−×
scu-MIR528Saccharum cultivarJXQF01030556.1:160~293,+×
sit-MIR528Setaria italica (L.) P. BeauvoisScaffold_9:57157041~57157170,−SETIT_040610mg
sbi-MIR528Sorghum bicolor (L.) MoenchChr1:79165414~79165537,−sbi-MIR528
spo-MIR528Spirodela polyrhiza (L.) Schleid.Pseudo17:196564~196705,−×
tae-MIR528aTriticum aestivum L.Chr4B:162371969~162372094,+×
tae-MIR528bTriticum aestivum L.Chr4D:32161888~32162013,+×
tae-MIR528cTriticum aestivum L.Chr5A:210090100~210090222,+×
zma-MIR528aZea mays L.Chr1:6409229~6409390,+zma-MIR528a
zma-MIR528bZea mays L.Chr9:153752320~153752436,−zma-MIR528b
a ‘+’ and ‘−’ indicate the forward and reverse strands, respectively. b ‘×’ indicates nonexistence.
Table 2. MiR528-regulated agronomic traits and potential applications in monocot crops.
Table 2. MiR528-regulated agronomic traits and potential applications in monocot crops.
Regulated TraitCrop SpeciesExperimental ApproachRegulatory Mechanism and Trait OutcomesPotential ApplicationReference
Flowering timeRiceOverexpressionRepresses OsRFI2 via OsSPL7 activation, promoting early flowering under long-day conditionsPhotoperiod adaptation and regional cultivation optimization [14]
Plant architectureCreeping bentgrassTransgenic overexpression of pri-osa-miR528Reduces internode length, increases tillering and vascular bundles, and enhances lodging resistanceBreeding for lodging-resistant cultivars [16]
Seed developmentRiceExpression profilingTargets copper-binding proteins and ascorbate oxidases during grain fillingEnhancing grain development and seed vigor [45,46]
Embryogenic developmentMaizeFunctional validationRegulates SOD, PLC, and transcription factors to promote somatic embryogenesisOptimization of somatic embryogenesis and plant regeneration protocols [48,49]
Biotic stress (viral)RiceMutant analysisAGO18 sequestration de-represses AO, increasing ROS-mediated antiviral defenseBreeding virus-resistant varieties [18,54]
Biotic stress (insect)RiceExpression analysislncRNA-mediated AO de-repression activates ascorbate defense against brown planthopperDevelopment of brown planthopper-resistant cultivars [56]
Biotic stress (fungal)MaizeCross-kingdom studyTargets fungal FvTTP to reduce mycotoxins and enhance SA signalingBreeding Fusarium-resistant maize with reduced mycotoxin accumulation [57]
Salt stressCreeping bentgrassTransgenic overexpression of pri-osa-miR528Suppresses AAO and CBP1, improving ion homeostasisBreeding salt-tolerant cultivars [16]
Salt stressRiceOverexpressionAO suppression elevates AsA/ABA, enhancing osmotic adjustment and ROS scavengingImproving salt tolerance via ABA–AsA–ROS modulation [59]
Temperature stress (cold)BananaMelatonin treatmentMaPPOs repression reduces enzymatic browning, enhances antioxidant activityPostharvest preservation via PPO suppression [61]
Temperature stress (cold)RiceCo-overexpressionSynergistic action with miR397/miR408 reduces oxidative damageDeveloping cold-tolerant rice through miRNA synergy [62]
Water stress (drought)WheatExpression profilingDownregulation coordinates drought-responsive gene networksBreeding drought-resilient wheat cultivars [20]
Water stress (drought)RiceOverexpressionIncreases IAA accumulation and reduces ROS, promoting root elongationEnhancing drought tolerance via IAA and ROS regulation [66]
Arsenic (As III) stressRiceOverexpressionSuppresses CBP and IAR1, disrupting As uptake and antioxidant defenseEnhancing arsenic tolerance and reducing arsenic accumulation [21]
Heavy-metal stress (Al)MaizeTissue-specific profilingMediates root-specific responses to aluminum toxicityEnhancing aluminum tolerance in maize via root-specific regulation [23]
Heavy-metal stress (Cd)RiceRegulatory module analysisWRKY51-miR528-UCL23 axis coordinates ROS homeostasis under cadmium stressMitigating cadmium toxicity by modulating ROS homeostasis [78]
Nitrogen homeostasisMaizeFunctional characterizationTargets ZmLAC3 to modify lignin biosynthesis and Casparian strip formationImproving nitrogen use efficiency [81,82]
Nitrogen homeostasisCreeping bentgrassTransgenic overexpression of pri-osa-miR528Enhances N assimilation via increased NiR activity and chlorophyll synthesisEnhancing nitrogen assimilation and growth under deficiency [16]
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Fu, H.; Zhang, L.; Hu, Y.; Liu, Z.; Wang, Z.; Shen, F.; Wang, W. MicroRNA528 and Its Regulatory Roles in Monocotyledonous Plants. Int. J. Mol. Sci. 2025, 26, 7334. https://doi.org/10.3390/ijms26157334

AMA Style

Fu H, Zhang L, Hu Y, Liu Z, Wang Z, Shen F, Wang W. MicroRNA528 and Its Regulatory Roles in Monocotyledonous Plants. International Journal of Molecular Sciences. 2025; 26(15):7334. https://doi.org/10.3390/ijms26157334

Chicago/Turabian Style

Fu, Hailin, Liwei Zhang, Yulin Hu, Ziyi Liu, Zhenyu Wang, Fafu Shen, and Wei Wang. 2025. "MicroRNA528 and Its Regulatory Roles in Monocotyledonous Plants" International Journal of Molecular Sciences 26, no. 15: 7334. https://doi.org/10.3390/ijms26157334

APA Style

Fu, H., Zhang, L., Hu, Y., Liu, Z., Wang, Z., Shen, F., & Wang, W. (2025). MicroRNA528 and Its Regulatory Roles in Monocotyledonous Plants. International Journal of Molecular Sciences, 26(15), 7334. https://doi.org/10.3390/ijms26157334

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop