1. Introduction
Myrica rubra, commonly known as Chinese bayberry, is a subtropical evergreen tree in the Myricaceae family that is native to southern China. Its fruit, prized for its distinctive sweet-and-sour flavor and high nutrimental value, is widely favored by consumers and makes a substantial contribution to the local economy [1]. Beyond its commercial appeal, Myrica rubra has attracted increasing scientific attention due to its potential health-promoting properties [2]. Previous studies have demonstrated that its fruit contains abundant bioactive compounds, such as anthocyanins, ascorbic acid, phenolic acids, and flavonols, which exhibit strong antioxidant, anti-inflammatory, antibacterial, and antitumor activities. These bioactive constituents are believed to contribute to the therapeutic potential of Myrica rubra, making it a promising candidate for further pharmacological and nutraceutical investigations [3,4,5]. The ripening period of Myrica rubra generally ranges from late May to early July, varying according to cultivar and geographical location. Due to its relatively short maturation window, optimal flavor is achieved only when the fruit is harvested at full ripeness. However, improper postharvest handling often leads to rapid softening and decay, resulting in substantial economic losses. Therefore, understanding the molecular mechanisms underlying fruit ripening in Myrica rubra is crucial for both improving postharvest management strategies and enhancing the commercial value of this species.
Fruit ripening is a complex physiological process regulated by diverse internal and external factors, including temperature, light, nutrients, hormones, epigenetic regulators, and endogenous signaling pathways [6,7]. Among these factors, ethylene is the best-characterized and most direct regulator of the fruit ripening process [8]. Ethylene, a gaseous plant hormone, governs a wide range of developmental processes, such as seed germination, fruit maturation, organ senescence, and stress responses [9,10,11]. The ethylene biosynthesis pathway is relatively simple, involving two key enzymatic steps. First, S-adenosyl-L-methionine (SAM) is converted into 1-aminocyclopropane-1-carboxylic acid (ACC) by ACC synthase (ACS), which catalyzes the rate-limiting step and serves as a key regulatory point in ethylene production [12,13]. Next, ACC is converted into ethylene by ACC oxidase (ACO). ACS genes play pivotal roles in ripening regulation across diverse species. For instance, PpACS1a is crucial for pear ripening, showing increased expression during the ripening stage [14]. ACS1 plays a dominant role in blueberry ripening, with cultivar-specific expression patterns [15]. In octoploid strawberry plants, FaACS27 and FaACS29 are highly expressed during achene development and contribute to ripening regulation [16]. In apples, MdACS3a regulates ripening, while MdACS6 acts at earlier developmental stages [17,18]. In tomatoes, RNA interference (RNAi) targeting ACS6, ACS1, and ACS2 suppresses ethylene biosynthesis, delays ripening, and extends shelf life [19]. In bananas, a multilayered regulatory cascade involving MaXB3, MaNAC, and MaERF11 modulates ripening, wherein MaXB3 negatively regulates ethylene biosynthesis by promoting the degradation of MaNAC2, MaACS1, and MaACO1 [20].
Beyond fruit ripening, ACS genes play essential roles in various aspects of plant growth and development by regulating ethylene biosynthesis. These roles include seed germination, root and shoot elongation, floral organ development, fruit set, leaf senescence, and organ abscission [21,22]. In Arabidopsis, AtACS7 is highly expressed during seed germination and contributes to early seedling development [23]. Mutations in AtACS4 and AtACS8 reduce ethylene production and affect lateral root formation [24]. AtACS2 and AtACS6 participate in petal and stamen development, with mutations resulting in floral defects or infertility [25]. In tomato plants, SlACS2 plays a critical role in floral organ development and normal fruit set, while SlACS2 and SlACS4 are key regulators of fruit softening and color change during ripening [26]. In the Cucurbitaceae, ACS genes are involved in sex determination. For example, CmACS6, CmACS7, CmACS9, and CmACS11 in melon, and ClACS1, ClACS7, and ClACS11 in watermelon, are predominantly expressed in female flowers, while ClACS6 shows high levels of expression in male flowers [27]. In Chinese chestnut (Castanea mollissima), CmACS7 expression determines ovule fertility, and its overexpression prior to fertilization leads to excessive ACC accumulation and ovule abortion [28]. Moreover, a natural allelic variant of ACS7 in watermelon plants modulates primary root elongation through ethylene-mediated signaling [29]. Transcriptional regulation of the ACS genes by various transcription factors has also been reported. In Arabidopsis, WRKY22 promotes ethylene biosynthesis by activating ACS5 and ACO5, thereby modulating root development [30], while WRKY71 regulates ethylene-mediated leaf senescence by activating EIN2, ORE1, and ACS2 [31].
Increasing research evidence shows that ACS genes are responsive to both biotic and abiotic stresses, thereby modulating ethylene production to enhance stress adaptation [32,33]. In rice, OsACS5 is strongly upregulated under hypoxic stress, promoting adventitious root formation through ethylene accumulation [34]. In bananas, MaACS1 and MaACS14 display distinct expression patterns under low-potassium and low-nitrogen stress [35]. Elsewhere, the ethylene response factor GsERF1 improves aluminum tolerance in Arabidopsis by upregulating ACS4, ACS5, and ACS6 [36]. Similarly, glutathione enhances resistance to necrotrophic pathogens and salt stress by triggering the WRKY33-mediated activation of ACS2 and ACS6 [37]. In cotton plants, the expression patterns of GhACS10 and GhACS12 change under various abiotic stresses, including cold, heat, drought, and salinity [38]. In sugarcane, ACS2 and ACS3 respond to low-nitrogen stress by regulating ethylene biosynthesis, contributing to stress tolerance and sugar accumulation [39]. These findings highlight the central role of ACS genes in regulating ethylene signaling in response to environmental stimuli.
Given the importance of the ACS genes in ethylene biosynthesis and their diverse roles in plant development and stress adaptation, investigating their structure, evolution, and expression patterns in various plants is essential. Myrica rubra genome sequencing was recently completed and published. However, the ACS gene family of this species has still not been reported. For this study, we performed a genome-wide identification of the ACS gene family in Myrica rubra and investigated their phylogenetic relationships, gene structures, conserved motifs, and chromosomal distributions. In addition, synteny analysis was conducted to evaluate evolutionary conservation, and gene expression profiles were analyzed to explore their potential roles during fruit development. These findings lay a solid foundation for future studies on ethylene biosynthesis and fruit ripening in Myrica rubra and related species.
2. Results
2.1. Identification of the ACS Gene Family in Myrica rubra
A total of seven MrACS gene family members in Myrica rubra were identified using a combination of BLAST (version 2.16.0) and HMMER (version 3.4) searches (Table 1 and Table S1). These proteins range in length from 446 amino acids (MrACS3) to 551 amino acids (MrACS7), with molecular weights (MW) of between 50.18 kDa and 60.34 kDa. The isoelectric points (pI) vary from 5.84 (MrACS2) to 8.33 (MrACS7), suggesting differences in protein charge. The aliphatic index for these proteins ranged from 82.6 to 89.6, and the grand average of hydropathicity (GRAVY) values was consistently negative, indicating that the MrACS proteins are hydrophilic. Subcellular localization predictions revealed that five of the MrACS proteins were localized in the cytoplasm, while two were localized in the nucleus. Additionally, the predicted tertiary structures showed that all MrACS proteins are composed of abundant α-helices and β-sheets, exhibiting overall compact three-dimensional structures (Figure 1).
2.2. Phylogenetic Analysis of the MrACS Gene Family
To access the evolutionary relationships among ACS genes, a phylogenetic tree was constructed based on the full-length protein sequences from seven plant species, including Myrica rubra (7 MrACSs), Arabidopsis thaliana (7 AtACSs), Malus domestica (7 MdACSs), Prunus persica (6 PpACSs), Solanum lycopersicum (13 SlACSs), Cucurbita pepo (9 CpACSs), and Vitis vinifera (6 VvACSs). A total of 55 ACS protein sequences were aligned and analyzed using MEGA 7.0 software (Supplementary Table S2). The resulting phylogenetic tree revealed that the ACS gene family could be classified into three major clades. The largest clade comprised 42 members, which were further subdivided into two main subgroups (Clade III and Clade IV). Clade I and Clade II contained 3 and 10 members, respectively, each also forming two subgroups (Figure 2). MrACS genes were distributed across different clades, with several clustering closely with homologs from the Rosaceae species such as peach and apple, suggesting a shared ancestry or conserved selection pressure.
2.3. Gene Structure and Conserved Motif Analysis of MrACSs
To further investigate the structural characteristics and evolutionary relationships of the MrACS gene family, conserved domains, protein motifs, and gene structures were analyzed using NCBI Batch CD-search (Bethesda, Maryland, USA), MEME (Knoxville, Tennessee, USA), and TBtools (Guangzhou, China). All MrACS proteins, except for MrACS3, contained the conserved PLN02450 domain (Figure 3), which is characteristic of the ACS protein family. The gene structure analysis revealed that MrACS genes contain between three and four exons, indicating relatively conserved exon-intron organization. Ten conserved motifs were identified among the MrACS proteins. Notably, MrACS1 and MrACS2 were each missing one motif, whereas the remaining members shared nine common motifs (Supplementary Table S3). These findings, in conjunction with the identified phylogenetic relationships, suggested that closely related members within the same clade exhibit highly similar motif compositions. This pattern supports the hypothesis that ACS subfamilies may possess conserved functional roles.
2.4. Chromosomal Localization and Collinearity Analysis of the MrACS Genes
The results of the chromosomal localization analysis showed that the seven MrACS genes were distributed on chromosomes 3, 4, 6, 7, and 8, with MrACS4 and MrACS7 both located on chromosome 4, while MrACS1 was located on an unanchored scaffold (Figure 4). In addition, the collinearity analysis revealed that two gene duplication events were identified: MrACS4–MrACS5 and MrACS6–MrACS7 (Figure 5). Ka/Ks ratio analysis showed values of 0.09 and 0.16 for these gene pairs, respectively (Table 2), indicating that the purifying selection process has acted to conserve their functions.
To further explore the evolutionary conservation and divergence of the MrACS gene family, synteny analyses were conducted with three fruit crops (comprising Malus domestica, Vitis vinifera, and Prunus persica) and two vegetable crops (comprising Cucurbita pepo and Solanum lycopersicum). The results revealed 10 homologous gene pairs between bayberry and tomato, 9 pairs with grape and peach, and 6 and 8 pairs with apple and pumpkin, respectively (Figure 6). These syntenic relationships suggest that the ACS genes in these species may share a common ancestral origin and that gene duplication or conservation events likely occurred after the divergence of these plant lineages.
2.5. Expression Patterns of MrACS Genes in Myrica rubra
To investigate the expression profiles of MrACS genes, transcripts per million (TPM) values were calculated (Table S4), based on the RNA-seq data in Myrica rubra, at three time points of 57, 85, and 113 days after pollination (DAP). The expression levels of three genes (MrACS5, MrACS6, and MrACS7) increased with the increase in pollination days, while the expression levels of the other four MrACS genes remained unchanged (Figure 7a). To further explore the tissue-specific expression of the MrACS genes, the expression levels in Myrica rubra in the flowers, leaves, stems, and fruits were examined (Figure 7b). MrACS1, MrACS2, and MrACS3 exhibited consistently low expression across all tissues, indicating limited or possibly redundant roles in Myrica rubra. In contrast, the remaining four genes showed variable and tissue-specific expression patterns. MrACS7 was highly expressed in all tissues, with higher levels than MrACS6, implying a more ubiquitous regulatory function. MrACS4 showed low expression in flowers, whereas MrACS5 was specifically and highly expressed in fruit, with minimal expression in other tissues. These expression profiles suggest that MrACS genes may be functionally diversified and may play distinct roles during fruit development and in different tissues of Myrica rubra.
3. Discussion
Ethylene is a key regulator in plant growth, development, fruit ripening, and stress tolerance [9,40]. The enzyme 1-aminocyclopropane-1-carboxylate synthase (ACS) catalyzes the rate-limiting step in ethylene biosynthesis and is encoded by a multigene family [13]. In Myrica rubra, we identified seven ACS genes, providing insight into the genetic basis of ethylene production in this economically and ecologically important species. Gene structure analysis revealed conserved exon-intron organization among the MrACS genes, which is consistent with observations in other species such as Malus domestica and Prunus persica [17,41]. Chromosomal localization analysis showed that the MrACS genes are unevenly distributed across chromosomes, a pattern that is frequently observed in plant genomes and is often associated with segmental duplications or genome rearrangements [42]. In this study, the gene pairs MrACS4–MrACS5 and MrACS6–MrACS7 exhibited low Ka/Ks ratios, indicating that the purifying selection process had maintained their functional integrity. This suggests that these duplicated genes are under evolutionary constraints that preserve their roles in ethylene biosynthesis. Phylogenetic analysis confirmed that these MrACS genes are highly conserved and are closely related to ACS genes from members of the Rosaceae family, implying shared evolutionary pressures, particularly those related to fruit development and ripening [43]. Collinearity analysis also provided compelling evidence of conserved homologous relationships between Myrica rubra and other plant species, particularly Vitis vinifera and Prunus persica, highlighting the functional significance of ACS genes in ethylene-mediated physiological processes across diverse fruit-bearing species. The distribution of ACS genes across multiple chromosomes in Myrica rubra suggests that gene duplication events, including both segmental and tandem duplications, have likely played a pivotal role in the expansion of this gene family. Such duplication events have been widely documented in other plant species, where they contribute to the functional diversification and adaptive evolution of ethylene biosynthesis pathways [44]. The uneven distribution of MrACS genes across the Myrica rubra chromosomes indicates that their genomic arrangement is independent of chromosome size. Similar patterns have been observed in Arabidopsis thaliana and Oryza sativa, where gene density and the chromosomal positioning of ACS genes are shaped by evolutionary constraints and regulatory interactions, rather than genome size alone [45]. Further comparative genomic analyses, especially those focusing on gene duplication mechanisms and the role of regulatory interactions, could offer deeper insights into the evolutionary forces shaping the distribution and functional diversification of the ACS gene family in Myrica rubra.
Tissue-specific expression patterns of ACS genes often reflect functional divergence following gene duplication. In Myrica rubra, RT-qPCR analysis revealed that MrACS5 was predominantly expressed in fruit tissue, which is consistent with a specialized role in ripening-related ethylene production, similar to SlACS2/4 in tomatoes and MdACS1 in apples [46,47,48,49]. Conversely, MrACS1-3 exhibited uniformly low expression levels across tissues, possibly indicating low basal activity or inducibility under specific conditions, as seen with AtACS6 in Arabidopsis thaliana [50]. MrACS7 showed ubiquitous expression across all tissues, implying a conserved housekeeping role in basal ethylene biosynthesis. MrACS4 was moderately expressed in the vegetative organs and may be involved in processes like development or senescence. Comparable findings have been reported in Cucurbita maxima, where ethylene biosynthesis genes like CmaACS4, CmaACS7, and CmaACS9 display flower-type-specific expression, reinforcing the notion of functional divergence within the gene family [33]. These distinct expression patterns underscore the regulatory specialization of MrACS genes, which is potentially governed by promoter architecture, epigenetic modifications, and tissue-specific transcription factor interactions. Further mechanistic studies are essential to elucidate how these regulatory networks shape the spatiotemporal roles of MrACS genes in Myrica rubra.
Fruit ripening is a genetically programmed and hormone-regulated process, with ethylene playing a central role in climacteric fruits such as tomatoes and apples [51,52,53]. In tomatoes, the transcription factor NAC-like1 can bind to the promoters of the ethylene biosynthesis genes SlACS2 and SlACS4, thereby promoting ethylene production [47]. Additionally, in RIN- and NOR-deficient tomato mutants, the accumulation of ACS and ACO mRNAs is significantly suppressed, resulting in delayed fruit ripening [54,55]. In apples, MdERF2 represses MdACS1 transcription, and MdACS3 mutations extend shelf life [17,56]. Compared with these model systems, the molecular regulation of fruit ripening in Myrica rubra remains largely unexplored. However, the present study identified MrACS5 as a gene that is highly expressed during fruit development, suggesting its involvement in ethylene biosynthesis and fruit ripening. Unlike tomatoes or bananas, where multiple ACS genes are simultaneously activated during ripening, MrACS5 exhibited tissue- and stage-specific expression, indicating a potentially distinct and more specialized regulatory mechanism. Notably, the weak and constitutive expression of other MrACS genes implies that they may function under specific environmental stimuli or hormonal cues, rather than in general developmental regulation.
Beyond plant development, ACS genes are known to mediate responses to biotic and abiotic stresses. Ethylene, synthesized via the ACS pathways, functions as a critical signaling molecule in stress adaptation [57,58]. In Arabidopsis thaliana, for example, AtACS8 mediates Cu2+-induced ethylene production and contributes to pathogen defense [59], while AtACS4/5/6 genes are upregulated by GsERF1 under aluminum stress [36]. Pathogen-induced ethylene biosynthesis involves AtACS2/6 activation via MPK3/6 phosphorylation, linking MAPK cascades to hormone regulation [60]. In rice, the OsACS1/2 genes respond dynamically to flooding and hypoxia, supporting adaptive ethylene synthesis [61]. Ethylene also interacts with SA and JA signaling to fine-tune immune responses under biotic stress [62]. In kiwifruit, ASA suppresses ethylene biosynthesis by repressing AdACS1/2 transcription and destabilizing AdACS3 through the inhibition of AdMPK16-mediated phosphorylation, revealing multi-level regulation [63]. Although ACS genes are known to mediate stress responses in model plants, their roles in Myrica rubra remain unclear. Our results have revealed the tissue-specific expression of MrACS genes, but their response to stress has not yet been explored. Future studies should investigate their stress inducibility and potential involvement in ethylene-mediated adaptation. In addition, the possible link between ethylene biosynthesis and bioactive compound regulation in Myrica rubra merits further research.
4. Materials and Methods
4.1. Plant Materials
Myrica rubra was cultivated in an orchard located in Wenling City, Zhejiang Province, China. Trees with similar growth potential and identical environmental conditions were selected for the study. Samples were collected from all tissues of Myrica rubra, with at least three biological replicates per sample. All samples were immediately snap-frozen in liquid nitrogen and stored at −80 °C for subsequent analysis.
4.2. Identification of ACS Genes in Myrica rubra
The methodology followed in this study was based on the approach outlined by Xie et al. [64]. The genome sequence, assembly files, and annotation information of Myrica rubra were retrieved from the Myrica rubra database (
4.3. Phylogenetic Analysis of ACS Genes in Myrica rubra
The ACS protein sequences of Arabidopsis thaliana, pumpkin (Cucurbita pepo), apple (Malus domestica), peach (Prunus persica), tomato (Solanum lycopersicum), grape (Vitis vinifera), and bayberry (Myrica rubra) were aligned multiple times using MEGA software, and a phylogenetic tree was constructed using the neighbor-joining (NJ) method.
4.4. Chromosome Distribution and Collinearity Analysis of ACS Genes in Myrica rubra
Genomic data for these species were downloaded from the NCBI website (
4.5. Analysis of ACS Gene Structure and Conserved Motifs in Myrica rubra
The exon-intron structure of the ACS gene was visualized using TBtools. Conserved motif analysis was conducted using the MEME suite (
4.6. Expression Profiling of MrACS Genes via Transcriptome Analysis and Quantitative Real-Time PCR
Transcriptome data from 57, 85, and 113 days after pollination (DAP) were selected to represent key stages of Myrica rubra fruit development—namely, immature, color-break, and fully ripe stages—during which ethylene biosynthesis and sugar accumulation are actively modulated [70]. The associated RNA-seq accession numbers are provided in Table S5 and are publicly available via the NCBI.
Total RNA was extracted using the Fast Pure Universal Plant Total RNA Extraction Kit (Vazyme, Nanjing, China), according to the manufacturer’s protocol. The RNA purity and concentration were analyzed using Nanodrop2000, and RNA integrity was assessed by 2% agarose gel electrophoresis. cDNA was performed following the manufacturer’s instructions by Prime Script™RT Kit (one-step gDNA removal) (Takara, Shiga, Japan). Specific primers for qRT-PCR were designed using Primer Premier v.5.0. SYBR Green PCR was carried out using a qTOWER3G fluorescence quantitative PCR instrument (Jena, Germany). In each reaction, 1 µL of the cDNA template, 10 µL of Taq Pro universal SYBR qPCR Master Mix (Vazyme, Nanjing, China), and 0.2 µm of specific primers were mixed, and ddH2O was added to a volume of 20 µL. The reaction was pre-denatured at 95 °C for 10 s, followed by 40 cycles of amplification at 95 °C for 10 s and 60 °C for 30 s. Three biological replicates and three technical replicates were performed for all reactions, using MrACT as an internal reference gene. Threshold cycle (Ct) values were automatically determined using a qTOWER3G fluorescence quantitative PCR instrument. The relative gene expression was calculated using the 2−ΔΔCT method [71]. The specific primers involved in this study are shown in Supplementary Table S6.
4.7. Statistical Analysis
GraphPad Prism 9 was used for data analysis and graphing. The results are expressed as the mean ± standard deviation (SD) from at least three replicate experiments. A one-way ANOVA, followed by Tukey’s test, was used to determine whether there was a statistically significant difference in the different independent groups.
5. Conclusions
In this study, seven ACS genes (MrACS1-7) were identified in Myrica rubra and were characterized by conserved structural features, including the PLN02450 domain (except MrACS3), hydrophilic properties, and α/β-rich tertiary structures. Phylogenetic analysis grouped these genes into three clades, revealing evolutionary conservation with Rosaceae species (Prunus persica and Malus domestica). Segmental duplication events (MrACS4-MrACS5 and MrACS6-MrACS7) and low Ka/Ks ratios (0.09–0.16) suggest that the purifying selection process has contributed to the maintenance of functional stability. Tissue-specific expression profiling highlighted MrACS5 as a fruit-specific gene that is significantly upregulated during late developmental stages (57–113 DAP), suggesting its critical role in ethylene-mediated ripening. In contrast, MrACS7 exhibited broad expression across tissues, implying a regulatory function in basal ethylene metabolism. These findings establish a foundation for deciphering ethylene biosynthesis in Myrica rubra and prioritizing MrACS5 as a key candidate for further functional studies aimed at improving postharvest fruit quality.
W.J. and H.H. designed the experiment. X.L. identified and characterized the MYB gene family. H.H. performed the experiments. H.H., X.L. and Y.L. prepared the manuscript. W.J. and F.W. revised the manuscript. All authors have read and agreed to the published version of the manuscript.
Not applicable.
Plant materials from bayberry were treated and collected with permission from Wenling City, Zhejiang Province, China. The plant materials did not include any wild species at risk of extinction. We complied with the relevant institutional, national, and international guidelines and legislation for plant study.
The data presented in this study are available in the article and
We sincerely thank our colleagues and collaborators for their valuable insights and technical assistance throughout this study.
The authors declare no competing interests.
Footnotes
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.
Figure 1 Predicted tertiary structures of seven MrACS proteins. Blue regions represent α-helices, red regions indicate β-sheets, and yellow regions denote random coils.
Figure 2 Phylogenetic relationships of the ACS proteins in Myrica rubra and six other species. This neighbor-joining phylogenetic tree was constructed based on a multiple sequence alignment of 55 ACS protein sequences from seven species. Different colors represent different subfamilies (I–IV).
Figure 3 Phylogenetic analysis, gene structure, conserved domains, and conserved motifs of the MrACS gene family. (A) Phylogenetic tree of the MrACS proteins. (B) Conserved motif distribution of the MrACS proteins. Motifs 1–10 are shown as colored boxes, with different colors representing distinct motifs. (C) Conserved domain architecture of the MrACS proteins. (D) Exon-intron structures of the MrACS genes. Green boxes represent exons, and black lines represent introns.
Figure 4 Schematic representations for the chromosomal distribution of MrACS genes.
Figure 5 Distribution and duplication events of MrACS genes across the bayberry genome. The gray lines indicate synteny blocks in the poplar genome, and the red lines indicate the synteny gene pairs of the ACS gene family.
Figure 6 Synteny analysis of ACS genes between Myrica rubra and Cucurbita pepo, Solanum lycopersicum, Malus domestica, Vitis vinifera, and Prunus persica. Gray lines in the background indicate the collinear blocks within Myrica rubra and other plant genomes, and the red lines indicate the ACS gene pairs. The chromosomes of Myrica rubra are labeled in magenta, while chromosomes of the other species are labeled in green.
Figure 7 Expression patterns of MrACS genes in bayberry after pollination. (a) The heatmap shows the expression levels of MrACS genes on the 57th day, 85th day, and 113th day after pollination. (b) Relative expression levels of MrACS1-7 in the flower, leaf, stem, immature fruit, and ripe fruit. The results are expressed as the means ± SEM of three biological replicates. Different letters indicate extremely significant differences at p < 0.01, according to a one-way analysis of variance (ANOVA) with Tukey’s multiple comparisons test.
The characteristics of 7 ACSs in Myrica rubra.
| Gene Name | Number of | Molecular Weight (kDa) | Theoretical pI | Instability Index | Aliphatic Index | Grand Average of | Subcellular |
|---|---|---|---|---|---|---|---|
| MrACS1 | 470 | 52.77 | 8.22 | 45.56 | 83.21 | −0.269 | Cytoplasm |
| MrACS2 | 477 | 53.71 | 5.84 | 47.28 | 82.6 | −0.24 | Nucleus |
| MrACS3 | 446 | 50.18 | 5.79 | 50.83 | 83.77 | −0.336 | Cytoplasm |
| MrACS4 | 486 | 54.60 | 6.6 | 43.83 | 82.84 | −0.18 | Cytoplasm |
| MrACS5 | 479 | 53.64 | 6.21 | 50.03 | 83.63 | −0.209 | Cytoplasm |
| MrACS6 | 528 | 58.41 | 8.4 | 50.54 | 89.6 | −0.084 | Nucleus |
| MrACS7 | 551 | 60.34 | 8.33 | 48.73 | 89 | −0.07 | Cytoplasm |
Ka/Ks of MrACS.
| Ka | Ks | Ka/Ks | ||
|---|---|---|---|---|
| MrACS4 | MrACS5 | 0.17033069606252893 | 1.9638562679829152 | 0.08673277104819707 |
| MrACS6 | MrACS7 | 0.23165164492644064 | 1.4133469890885786 | 0.16390288210528203 |
Supplementary Materials
The following supporting information can be downloaded at:
1. Ren, H.Y.; He, Y.H.; Qi, X.J.; Zheng, X.L.; Zhang, S.W.; Yu, Z.P.; Hu, F.R. The bayberry database: A multiomic database for Myrica rubra, an important fruit tree with medicinal value. BMC Plant Biol.; 2021; 21, 452. [DOI: https://dx.doi.org/10.1186/s12870-021-03232-x]
2. Zhang, S.W.; Yu, Z.P.; Sun, L.; Ren, H.Y.; Zheng, X.L.; Liang, S.M.; Qi, X.J. An overview of the nutritional value, health properties, and future challenges of Chinese bayberry. PeerJ.; 2022; 10, e13070. [DOI: https://dx.doi.org/10.7717/peerj.13070] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/35265403]
3. Xia, W.; Gong, E.S.; Lin, Y.Y.; Li, T.; Lian, F.L.; Zheng, B.S.; Liu, R.H. Comparison of phytochemical profiles, antioxidant and antiproliferative activities in Chinese bayberry (Myrica rubra Sieb. et Zucc.) fruits. J. Food Sci.; 2021; 86, pp. 4691-4703. [DOI: https://dx.doi.org/10.1111/1750-3841.15899] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/34549442]
4. Wang, Y.S.; Chen, J.B.; Wang, Y.; Zheng, F.H.; Qu, M.Y.; Huang, Z.W.; Yan, J.L.; Bao, F.P.; Li, X.; Sun, C.D.
5. Zhang, Q.Z.; Huang, Z.J.; Wang, Y.; Wang, Y.B.; Fu, L.L.; Su, L.J. Chinese bayberry (Myrica rubra) phenolics mitigated protein glycoxidation and formation of advanced glycation end-products: A mechanistic investigation. Food Chem.; 2021; 361, 130102. [DOI: https://dx.doi.org/10.1016/j.foodchem.2021.130102] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/34029891]
6. Fenn, M.A.; Giovannoni, J.J. Phytohormones in fruit development and maturation. Plant J.; 2021; 105, pp. 446-458. [DOI: https://dx.doi.org/10.1111/tpj.15112]
7. Tang, D.G.; Gallusci, P.P.; Lang, Z.B. Fruit development and epigenetic modifications. New Phytol.; 2020; 228, pp. 839-844. [DOI: https://dx.doi.org/10.1111/nph.16724]
8. Zhang, Y.Q.; Berman, A.; Shani, E. Plant hormone transport and localization: Signaling molecules on the move. Annu. Rev. Plant Biol.; 2023; 22, pp. 453-479. [DOI: https://dx.doi.org/10.1146/annurev-arplant-070722-015329]
9. Huang, J.Y.; Zhao, X.B.; Bürger, M.; Chory, J.; Wang, X.C. The role of ethylene in plant temperature stress response. Trends Plant Sci.; 2023; 28, pp. 808-824. [DOI: https://dx.doi.org/10.1016/j.tplants.2023.03.001]
10. Zhao, H.; Yin, C.C.; Ma, B.; Chen, S.Y.; Zhang, J.S. Ethylene signaling in rice and Arabidopsis: New regulators and mechanisms. J. Integr. Plant Biol.; 2021; 63, pp. 102-125. [DOI: https://dx.doi.org/10.1111/jipb.13028]
11. Yin, C.C.; Huang, Y.H.; Zhang, X.; Zhou, Y.; Chen, S.Y.; Zhang, J.S. Ethylene-mediated regulation of coleoptile elongation in rice seedlings. Plant Cell Environ.; 2023; 46, pp. 1060-1074. [DOI: https://dx.doi.org/10.1111/pce.14492]
12. Pattyn, J.; Vaughan-Hirsch, J.; Van de Poel, B. The regulation of ethylene biosynthesis: A complex multilevel control circuitry. New Phytol.; 2021; 229, pp. 770-782. [DOI: https://dx.doi.org/10.1111/nph.16873] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/32790878]
13. Booker, M.A.; DeLong, A. Producing the ethylene signal: Regulation and diversification of ethylene biosynthetic enzymes. Plant Physiol.; 2015; 169, pp. 42-50. [DOI: https://dx.doi.org/10.1104/pp.15.00672] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/26134162]
14. Shi, H.Y.; Zhang, Y.X. Expression and regulation of pear 1-aminocyclopropane-1-carboxylic acid synthase gene (PpACS1a) during fruit ripening, under salicylic acid and indole-3-acetic acid treatment, and in diseased fruit. Mol. Biol. Rep.; 2014; 41, pp. 4147-4154. [DOI: https://dx.doi.org/10.1007/s11033-014-3286-3] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/24562629]
15. Wang, Y.W.; Acharya, T.P.; Malladi, A.; Tsai, H.J.; NeSmith, D.S.; Doyle, J.W.; Nambeesan, S.U. Atypical Climacteric and Functional Ethylene Metabolism and Signaling During Fruit Ripening in Blueberry (Vaccinium sp.). Front. Plant Sci.; 2022; 23, 932642. [DOI: https://dx.doi.org/10.3389/fpls.2022.932642]
16. Upadhyay, R.K.; Motyka, V.; Pokorna, E.; Dobrev, P.I.; Lacek, J.; Shao, J.; Lewers, K.S.; Mattoo, A.K. Comprehensive profiling of endogenous phytohormones and expression analysis of 1-aminocyclopropane-1-carboxylic acid synthase gene family during fruit development and ripening in octoploid strawberry (Fragaria × ananassa). Plant Physiol. Biochem.; 2023; 196, pp. 186-196. [DOI: https://dx.doi.org/10.1016/j.plaphy.2023.01.031]
17. Wang, A.D.; Yamakake, J.; Kudo, H.; Wakasa, Y.; Hatsuyama, Y.; Igarashi, M.; Kasai, A.; Li, T.; Harada, T. Null mutation of the MdACS3 gene, coding for a ripening-specific 1-aminocyclopropane-1-carboxylate synthase, leads to long shelf life in apple fruit. Plant Physiol.; 2009; 151, pp. 391-399. [DOI: https://dx.doi.org/10.1104/pp.109.135822]
18. Li, T.; Tan, D.M.; Liu, Z.; Jiang, Z.Y.; Wei, Y.; Zhang, L.C.; Li, X.Y.; Yuan, H.; Wang, A.D. Apple MdACS6 regulates ethylene biosynthesis during fruit development involving ethylene-responsive factor. Plant Cell Physiol.; 2015; 56, pp. 1909-1917. [DOI: https://dx.doi.org/10.1093/pcp/pcv111]
19. Gupta, A.; Pal, R.K.; Rajam, M.V. Delayed ripening and improved fruit processing quality in tomato by RNAi-mediated silencing of three homologs of 1-aminopropane-1-carboxylate synthase gene. Plant Physiol.; 2013; 170, pp. 987-995. [DOI: https://dx.doi.org/10.1016/j.jplph.2013.02.003]
20. Shan, W.; Kuang, J.F.; Wei, W.; Fan, Z.Q.; Deng, W.; Li, Z.G.; Bouzayen, M.; Pirrello, J.; Lu, W.J.; Chen, J.Y. MaXB3 modulates MaNAC2, MaACS1, and MaACO1 stability to repress ethylene biosynthesis during banana fruit ripening. Plant Physiol.; 2020; 184, pp. 1153-1171. [DOI: https://dx.doi.org/10.1104/pp.20.00313]
21. Park, C.; Lee, H.Y.; Yoon, G.M. The regulation of ACC synthase protein turnover: A rapid route for modulating plant development and stress responses. Curr. Opin. Plant Biol.; 2021; 63, 102046. [DOI: https://dx.doi.org/10.1016/j.pbi.2021.102046] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/33965697]
22. Li, J.Y.; Cheng, K.; Lu, Y.; Wen, H.Y.; Ma, L.Q.; Zhang, C.J.; Suprun, A.R.; Zhu, H.L. Regulation of 1-aminocyclopropane-1-carboxylic acid synthase (ACS) expression and its functions in plant life. Plant Horm.; 2025; 1, e002. [DOI: https://dx.doi.org/10.48130/ph-0025-0002]
23. Tang, X.L.; Liu, R.; Mei, Y.Y.; Wang, D.; He, K.X.; Wang, N.N. Identification of key ubiquitination sites involved in the proteasomal degradation of AtACS7 in Arabidopsis. Int. J. Mol. Sci.; 2024; 25, 2931. [DOI: https://dx.doi.org/10.3390/ijms25052931] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/38474174]
24. Joo, S.; Seo, Y.S.; Kim, S.M.; Hong, D.K.; Park, K.Y.; Kim, W.T. Brassinosteroid induction of AtACS4 encoding an auxin-responsive 1-aminocyclopropane-1-carboxylate synthase 4 in Arabidopsis seedlings. Physiologia Plantarum.; 2006; 126, pp. 453-461. [DOI: https://dx.doi.org/10.1111/j.1399-3054.2005.00602.x]
25. Wang, N.N.; Shih, M.C.; Li, N. The GUS reporter-aided analysis of the promoter activities of Arabidopsis ACC synthase genes AtACS4, AtACS5, and AtACS7 induced by hormones and stresses. J. Exp. Bot.; 2005; 56, pp. 909-920. [DOI: https://dx.doi.org/10.1093/jxb/eri083]
26. Cao, H.H.; Chen, J.; Yue, M.; Xu, C.; Jian, W.; Liu, Y.D.; Song, B.Q.; Gao, Y.Q.; Cheng, Y.L.; Li, Z.G. Tomato transcriptional repressor MYB70 directly regulates ethylene-dependent fruit ripening. Plant J.; 2020; 104, pp. 1568-1581. [DOI: https://dx.doi.org/10.1111/tpj.15021]
27. Wang, Z.; Yadav, V.; Yan, X.; Cheng, D.; Wei, C.; Zhang, X. Systematic genome-wide analysis of the ethylene-responsive ACS gene family: Contributions to sex form differentiation and development in melon and watermelon. Gene.; 2021; 805, 145910. [DOI: https://dx.doi.org/10.1016/j.gene.2021.145910]
28. Cui, Y.; Ji, X.; Yu, W.; Liu, Y.; Bai, Q.; Su, S. Genome-Wide Characterization and Functional Validation of the ACS Gene Family in the Chestnut Reveals Its Regulatory Role in Ovule Development. Int. J. Mol. Sci.; 2024; 25, 4454. [DOI: https://dx.doi.org/10.3390/ijms25084454] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/38674037]
29. Mahmoud, A.; Qi, R.; Zhao, H.; Yang, H.; Liao, N.; Ali, A.; Malangisha, G.K.; Ma, Y.; Zhang, K.; Zhou, Y.
30. Wang, Z.; Wei, X.; Cui, X.; Wang, J.; Wang, Y.; Sun, M.; Zhao, P.; Yang, B.; Wang, Q.; Jiang, Y.Q. The transcription factor WRKY22 modulates ethylene biosynthesis and root development through transactivating the transcription of ACS5 and ACO5 in Arabidopsis. Physiol. Plant.; 2024; 176, e14371. [DOI: https://dx.doi.org/10.1111/ppl.14371]
31. Yu, Y.; Qi, Y.; Xu, J.; Dai, X.; Chen, J.; Dong, C.H.; Xiang, F. Arabidopsis WRKY71 regulates ethylene-mediated leaf senescence by directly activating EIN2, ORE1 and ACS2 genes. Plant J.; 2021; 107, pp. 1819-1836. [DOI: https://dx.doi.org/10.1111/tpj.15433] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/34296474]
32. Liu, S.Q.; Lei, C.; Zhu, Z.H.; Li, M.Z.; Chen, Z.P.; He, W.; Liu, B.; Chen, L.P.; Li, X.J.; Xie, Y.Z. Genome-wide analysis and identification of 1-aminocyclopropane-1-carboxylate synthase (ACS) gene family in wheat (Triticum aestivum L.). Int. J. Mol. Sci.; 2023; 24, 11158. [DOI: https://dx.doi.org/10.3390/ijms241311158] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/37446336]
33. Wang, C.J.; Li, W.L.; Chen, F.Y.; Cheng, Y.Q.; Huang, X.; Zou, B.X.; Wang, Y.L.; Xu, W.L.; Qu, S.P. Genome-wide identification and characterization of members of the ACS gene family in Cucurbita maxima and their transcriptional responses to the specific treatments. Int. J. Mol. Sci.; 2022; 23, 8476. [DOI: https://dx.doi.org/10.3390/ijms23158476]
34. Van Der Straeten, D.; Zhou, Z.; Prinsen, E.; Van Onckelen, H.A.; Van Montagu, M.C. A comparative molecular-physiological study of submergence response in lowland and deepwater rice. Plant Physiol.; 2001; 125, pp. 955-968. [DOI: https://dx.doi.org/10.1104/pp.125.2.955]
35. Tabassum, N.; Shafiq, M.; Fatima, S.; Tahir, S.; Tabassum, B.; Ali, Q.; Javed, M.A. Genome-wide in-silico analysis of ethylene biosynthesis gene family in Musa acuminata L. and their response under nutrient stress. Sci. Rep.; 2024; 14, 558. [DOI: https://dx.doi.org/10.1038/s41598-023-51075-3]
36. Li, L.; Li, X.; Yang, C.; Cheng, Y.; Cai, Z.; Nian, H.; Ma, Q. GsERF1 enhances Arabidopsis thaliana aluminum tolerance through an ethylene-mediated pathway. BMC Plant Biol.; 2022; 22, 258. [DOI: https://dx.doi.org/10.1186/s12870-022-03625-6]
37. Datta, R.; Kumar, D.; Sultana, A.; Hazra, S.; Bhattacharyya, D.; Chattopadhyay, S. Glutathione regulates 1-aminocyclopropane-1-carboxylate synthase transcription via WRKY33 and 1-aminocyclopropane-1-carboxylate oxidase by modulating messenger RNA stability to induce ethylene synthesis during stress. Plant Physiol.; 2015; 169, pp. 2963-2981. [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/26463088]
38. Wang, Z.; Li, F. Potential Roles of 1-Aminocyclopropane-1-carboxylic Acid Synthase Genes in the Response of Gossypium Species to Abiotic Stress by Genome-Wide Identification and Expression Analysis. Plants; 2022; 11, 1524. [DOI: https://dx.doi.org/10.3390/plants11111524]
39. Wu, Z.; Zhang, X.; Zhang, N.; Gao, X.; Feng, X.; Zeng, Q.; Chen, X.L.; Wu, J.Y.; Qi, Y.W. Identification of ACC synthetase genes in saccharum and their expression profiles during plant growth and in response to low-nitrogen stress. Trop. Plant Biol.; 2022; 15, pp. 197-210. [DOI: https://dx.doi.org/10.1007/s12042-022-09316-8]
40. Iqbal, N.; Trivellini, A.; Masood, A.; Ferrante, A.; Khan, N.A. Current understanding on ethylene signaling in plants: The influence of nutrient availability. Plant Physiol. Biochem.; 2013; 73, pp. 128-138. [DOI: https://dx.doi.org/10.1016/j.plaphy.2013.09.011]
41. Zeng, W.F.; Pan, L.; Liu, H.; Niu, L.; Lu, Z.H.; Cui, G.C.; Wang, Z.Q. Characterization of 1-aminocyclopropane-1-carboxylic acid synthase (ACS) genes during nectarine fruit development and ripening. Tree Genet. Genomes; 2015; 11, 18. [DOI: https://dx.doi.org/10.1007/s11295-015-0833-6]
42. Chen, H.T.; Bai, S.L.; Kusano, M.; Ezura, H.; Wang, N. Increased ACS enzyme dosage causes Initiation of climacteric ethylene production in tomato. Int. J. Mol. Sci.; 2022; 23, 10788. [DOI: https://dx.doi.org/10.3390/ijms231810788]
43. Zhang, Y.X.; Zhang, Y.R.; Yu, Z.; Wang, H.Y.; Ping, B.; Liu, Y.X.; Liang, J.K.; Ma, F.W.; Zou, Y.J.; Zhao, T. Insights into ACO genes across Rosaceae: Evolution, expression, and regulatory networks in fruit development. Genes. Genom.; 2024; 46, pp. 1209-1223. [DOI: https://dx.doi.org/10.1007/s13258-024-01551-5] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/39141243]
44. Zhang, T.C.; Qiao, Q.; Zhong, Y. Detecting adaptive evolution and functional divergence in aminocyclopropane-1-carboxylate synthase (ACS) gene family. Comput. Biol. Chem.; 2012; 38, pp. 10-16. [DOI: https://dx.doi.org/10.1016/j.compbiolchem.2012.04.001]
45. Ahmadizadeh, M.; Chen, J.T.; Hasanzadeh, S.; Ahmar, S.; Heidari, P. Insights into the genes involved in the ethylene biosynthesis pathway in Arabidopsis thaliana and Oryza sativa. J. Genet. Eng. Biotechnol.; 2020; 18, 62. [DOI: https://dx.doi.org/10.1186/s43141-020-00083-1]
46. Liu, Y.D.; Shi, Y.; Su, D.D.; Lu, W.; Li, Z. SlGRAS4 accelerates fruit ripening by regulating ethylene biosynthesis genes and SlMADS1 in tomato. Hortic. Res.; 2021; 8, 3. [DOI: https://dx.doi.org/10.1038/s41438-020-00431-9]
47. Gao, Y.; Wei, W.; Zhao, X.D.; Tan, X.L.; Fan, Z.Q.; Zhang, Y.P.; Jing, Y.; Meng, L.H.; Zhu, B.Z.; Zhu, H.L.
48. Dougherty, L.; Zhu, Y.D.; Xu, K.N. Assessing the allelotypic effect of two aminocyclopropane carboxylic acid synthase-encoding genes MdACS1 and MdACS3a on fruit ethylene production and softening in Malus. Hortic. Res.; 2016; 3, 16024. [DOI: https://dx.doi.org/10.1038/hortres.2016.24] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/27231553]
49. Sharma, K.; Gupta, S.; Sarma, S.; Rai, M.; Sreelakshmi, Y.; Sharma, R. Mutations in tomato 1-aminocyclopropane carboxylic acid synthase 2 uncover its role in development beside fruit ripening. Plant J.; 2021; 106, pp. 95-112. [DOI: https://dx.doi.org/10.1111/tpj.15148]
50. Kim, S.H.; Yang, S.H.; Kim, T.J.; Han, J.S.; Suh, J.W. Hypertonic stress increased extracellular ATP levels and the expression of stress-responsive genes in Arabidopsis thaliana seedlings. Biosci. Biotechnol. Biochem.; 2009; 73, pp. 1252-1256. [DOI: https://dx.doi.org/10.1271/bbb.80660]
51. Giovannoni, J.; Nguyen, C.; Ampofo, B.; Zhong, S.; Fei, Z. The epigenome and transcriptional dynamics of fruit ripening. Annu. Rev. Plant Biol.; 2017; 68, pp. 61-84. [DOI: https://dx.doi.org/10.1146/annurev-arplant-042916-040906]
52. Ji, Y.L.; Wang, A.D. Recent advances in epigenetic triggering of climacteric fruit ripening. Plant Physiol.; 2023; 192, pp. 1711-1717. [DOI: https://dx.doi.org/10.1093/plphys/kiad206] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/37002826]
53. Liu, M.C.; Pirrello, J.; Chervin, C.; Roustan, J.P.; Bouzayen, M. Ethylene control of fruit ripening: Revisiting the complex network of transcriptional regulation. Plant Physiol.; 2015; 169, pp. 2380-2390. [DOI: https://dx.doi.org/10.1104/pp.15.01361] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/26511917]
54. Ito, Y.; Nishizawa-Yokoi, A.; Endo, M.; Mikami, M.; Shima, Y.; Nakamura, N.; Kotake-Nara, E.; Kawasaki, S.; Toki, S. Re-evaluation of the rin mutation and the role of RIN in the induction of tomato ripening. Nat. Plants.; 2017; 3, pp. 866-874. [DOI: https://dx.doi.org/10.1038/s41477-017-0041-5]
55. Gao, Y.; Wei, W.; Fan, Z.Q.; Zhao, X.D.; Zhang, Y.P.; Jing, Y.; Zhu, B.Z.; Zhu, H.L.; Shan, W.; Chen, J.Y.
56. Li, T.; Jiang, Z.Y.; Zhang, L.C.; Tan, D.M.; Wei, Y.; Yuan, H.; Li, T.L.; Wang, A.D. Apple (Malus domestica) MdERF2 negatively affects ethylene biosynthesis during fruit ripening by suppressing MdACS1 transcription. Plant J.; 2016; 88, pp. 735-748. [DOI: https://dx.doi.org/10.1111/tpj.13289]
57. Shekhawat, K.; Fröhlich, K.; García-Ramírez, G.X.; Trapp, M.A.; Hirt, H. Ethylene: A master regulator of Plant-Microbe interactions under abiotic stresses. Cells; 2022; 12, 31. [DOI: https://dx.doi.org/10.3390/cells12010031]
58. Riyazuddin, R.; Verma, R.; Singh, K.; Nisha, N.; Keisham, M.; Bhati, K.K.; Kim, S.T.; Gupta, R. Ethylene: A master regulator of salinity stress tolerance in plants. Biomolecules; 2020; 10, 959. [DOI: https://dx.doi.org/10.3390/biom10060959]
59. Zhang, B.G.; Liu, H.F.; Ding, X.H.; Qiu, J.J.; Zhang, M.; Chu, Z. Arabidopsis thaliana ACS8 plays a crucial role in the early biosynthesis of ethylene elicited by Cu2+ ions. J. Cell Sci.; 2018; 131, jcs202424. [DOI: https://dx.doi.org/10.1242/jcs.202424]
60. Poulaki, E.G.; Tsolakidou, M.D.; Gkizi, D.; Pantelides, I.S.; Tjamos, S.E. The ethylene biosynthesis genes ACS2 and ACS6 modulate disease severity of Verticillium dahliae. Plants; 2020; 9, 907. [DOI: https://dx.doi.org/10.3390/plants9070907]
61. Zarembinski, T.I.; Theologis, A. Expression characteristics of OS-ACS1 and OS-ACS2, two members of the 1-aminocyclopropane-1-carboxylate synthase gene family in rice (Oryza sativa L. cv. Habiganj Aman II) during partial submergence. Plant Mol. Biol.; 1997; 33, pp. 71-77. [DOI: https://dx.doi.org/10.1023/B:PLAN.0000009693.26740.c3] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/9037160]
62. Yuan, M.; Huang, Y.Y.; Ge, W.N.; Jia, Z.H.; Song, S.S.; Zhang, L.; Huang, Y.L. Involvement of jasmonic acid, ethylene and salicylic acid signaling pathways behind the systemic resistance induced by Trichoderma longibrachiatum H9 in cucumber. BMC Genom.; 2019; 20, 144. [DOI: https://dx.doi.org/10.1186/s12864-019-5513-8]
63. Wang, J.; Liu, X.F.; Zhang, H.Q.; Allan, A.C.; Wang, W.Q.; Yin, X.R. Transcriptional and post-transcriptional regulation of ethylene biosynthesis by exogenous acetylsalicylic acid in kiwifruit. Hortic. Res.; 2022; 9, uhac116. [DOI: https://dx.doi.org/10.1093/hr/uhac116]
64. Xie, L.M.; Wang, Y.N.; Tao, Y.T.; Chen, L.X.; Lin, H.Y.; Qi, Z.C.; Li, J.M. Genome-wide identification and analysis of anthocyanin synthesis-related R2R3-MYB genes in Fragaria pentaphylla. BMC Genomics.; 2024; 25, 952. [DOI: https://dx.doi.org/10.1186/s12864-024-10882-2] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/39396954]
65. Ren, H.Y.; Yu, H.Y.; Zhang, S.W.; Liang, S.M.; Zheng, X.L.; Zhang, S.J.; Yao, P.; Zheng, H.K.; Qi, X.J. Genome sequencing provides insights into the evolution and antioxidant activity of Chinese bayberry. BMC Genom.; 2019; 20, 458. [DOI: https://dx.doi.org/10.1186/s12864-019-5818-7] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/31170907]
66. Garcia-Hernandez, M.; Berardini, T.Z.; Chen, G.H.; Crist, D.; Doyle, A.; Huala, E.; Knee, E.; Lambrecht, M.; Miller, N.; Mueller, L.A.
67. Mistry, J.; Chuguransky, S.; Williams, L.; Qureshi, M.; Salazar, G.A.; Sonnhammer, E.; Tosatto, S.; Paladin, L.; Raj, S.; Richardson, L.
68. Wilkins, M.R.; Gasteiger, E.; Bairoch, A.; Sanchez, J.C.; Williams, K.L.; Appel, R.D.; Hochstrasser, D.F. Protein identification and analysis tools in the ExPASy server. Methods Mol. Biol.; 1999; 112, pp. 531-552.
69. Bailey, T.L.; Boden, M.; Buske, F.A.; Frith, M.; Grant, C.E.; Clementi, L.; Ren, J.; Li, W.W.; Noble, W.S. MEME SUITE: Tools for motif discovery and searching. Nucleic Acids Res.; 2009; 37, pp. W202-W208. [DOI: https://dx.doi.org/10.1093/nar/gkp335]
70. Chen, X.; Shi, L.Y.; Shao, J.R.; Chen, W.; Zheng, Y.H.; Yang, Z.F. Molecular cloning and expression analysis of MrFRK2 in Chinese bayberry during fruit ripening. Acta Hortic. Sinica.; 2016; 43, 1585.
71. Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods; 2001; 25, pp. 402-408. [DOI: https://dx.doi.org/10.1006/meth.2001.1262] [PubMed: https://www.ncbi.nlm.nih.gov/pubmed/11846609]
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.
Details
1 Key Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China; [email protected] (H.H.); [email protected] (X.L.); [email protected] (Y.L.); [email protected] (F.W.), Shaoxing Academy of Biomedicine, Zhejiang Sci-Tech University, Shaoxing 312366, China
2 Key Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Science and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China; [email protected] (H.H.); [email protected] (X.L.); [email protected] (Y.L.); [email protected] (F.W.)