A new transcription factor, HgMYB60, regulates salt and drought stress of halophyte Halogeton glomeratus

A new transcription factor, HgMYB60, regulates salt and drought stress of halophyte Halogeton glomeratus | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 16 April 2025 V1 Latest version Share on A new transcription factor, HgMYB60, regulates salt and drought stress of halophyte Halogeton glomeratus Authors : Pengxu He [email protected] , Lirong Yao , Xiangling Sun , Lujuan Sun , Zhenghuang Zhang , Shangqing Hu , Yan Yan , … Show All … , Baochun Li , Hong Zhang , Xiaole Ma , Erjing Si , Ke Yang , Yaxiong Meng , Huajun Wang , and Juncheng Wang Show Fewer Authors Info & Affiliations https://doi.org/10.22541/au.174480048.89154542/v1 189 views 115 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Salt and drought represent major threats to global agricultural productivity and food security, but plants have developed a suite of adaptive responses conferring short-or longer-term survival to this stress. A new transcription factor HgMYB60 in H. glomeratus was explored to coordinate these responses. We cloned the full-length sequence of HgMYB60 , determined the expressions and functions of HgMYB60 , and explored the HgMYB60 assayed proteins related to sodium and potassium transport. The expression levels of HgMYB60 were increased in Arabidopsis mutants under salt and drought stress, and HgMYB60 -overexpressing Arabidopsis mutants showed enhanced survival to salt and drought stress. In addition, HgMYB60 mediated sodium ion efflux and potassium ion absorption at the cellular level. HgMYB60 was localized to the nucleus, and these results are thus consistent with a major role for Na + transport in the signaling systems that respond to salt and drought stress. 1 Introduction With the construction of standardized farmland, most crops are currently irrigated using shallow-buried drip irrigation, which can lead to increased salt and drought, especially in northwest China (Man et al. , 2024). Therefore, it is particularly important to cultivate resistant varieties. Abiotic stresses such as soil salinity and drought have a profoundly detrimental impact on the productivity and quality of globally significant economic crops (Devi et al. , 2012). Approximately 3.6 billion hectares of the world’s arable land are afflicted by soil degradation and salinization, resulting in annual global agricultural losses exceeding 12 billion US dollars due to salinity alone, while drought concurrently exerts a significant toll on crop yields (Shabala, 2013; Yang et al. , 2021). Both salinity anddrought significantly affect plant growth and development, physiological andbiochemical responses, and quality and yield. Consequently, enhancing the salt tolerance and drought resistance of crops is becoming a crucial strategy for managing extreme environmental conditions. In general, salt and drought stress disrupts various physiological mechanisms in plants, leading to changes in growth patterns (Angon et al. , 2022). When salts accumulate in the roots, plants experience physiological drought, which affects stomatal physiology, reduces photosynthesis, and inhibits plant growth. The growth of plant leaves slows down over time, and the leaves become smaller (James et al. , 2011; Kumar et al. , 2019; Shahzad et al. , 2019). Plants have evolved a series of intricate response mechanisms to adapt to salt stress, whichincludes defense against oxidative stress, prevention of ionic toxicity, and maintenance of the osmotic balance (Li, X et al. , 2020; Zhang et al. , 2020). Plant salt tolerance has been associated with various proteins and genes, including theplasma membrane cation/proton antiporter (SOS1), the vacuolar membrane Na + /H + exchanger (NHX), plasma membrane and vacuolar H + -ATPases, vacuolar H + -pyrophosphatases, potassium transporters, and other salt tolerance-related genes(Hamaji et al. , 2009). Plants enhance their tolerance to salt stress by reducing the accumulation of reactive oxygen species (ROS) through increased activity of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione S-transferase (GPX) (Gill & Tuteja, 2010; Hasanuzzaman et al. , 2019). Osmotic regulators such as proline and glycine betaine play a role in the osmotic adjustment of plants under salt stress (Sharma et al. , 2019).Under salt stress, the excessive accumulation of Na + in plants inhibits the absorption of K + , leading to a decrease in K + levels and disrupting the Na + /K + balance, which results in ionic toxicity. To reduce the Na + concentration in the cytoplasm, plants often employ compartmentalization to maintain ionic homeostasis and balance. Excessive Na + is allocated to older tissues or compartmentalized into vacuoles, enabling the plants to survive salinity (Zhu, 2003; Li, H et al. , 2020). Drought stress, similar to salt stress, disrupts plant nutritional homeostasis and photosynthetic efficiency, with water deficit in drought conditions leading to cellular turgor loss and consequent growth inhibition (Umair Hassan et al. , 2020; Razi & Muneer, 2021; Nardini, 2022). Drought also disrupts internal plantprocesses, such as carbon assimilation, increases oxidative damage, and alters leaf gas exchange, irreversibly affecting plant health (Nadeem et al. , 2019). Drought stress damages the plant cell membrane system, increasing ROS and creating an imbalance between ROS production and scavenging. This leads to membrane lipid peroxidation and oxidative stress in plant cells, elevating malondialdehyde (MDA) content and cell membrane permeability. The inactivation of proteins and enzymes in the plant cell membrane results in damage to the structure and function of biomembranes. Severe drought stress can lead to plant death. Plants mitigate drought-induced damage by enhancing antioxidant enzyme activity and transcriptionally regulating defense-related genes (Li et al. , 2022). Increasing crop productivity under stress conditions requires breakthroughs in drought-resistant breeding. However, traditional crops possess certain limitations in their stress tolerance. To address this issue, it is necessary to identify stress-tolerant genes from extreme stress-resistant plants. Transcription factors are essential proteins that regulate gene expression and biological functions by binding to cis-regulatory elements in the promoter regions of genes. MYB transcription factors were first discovered in viruses and represent one of the largest families of transcription factors, being nearly ubiquitous among all eukaryotes (Klempnauer et al. , 1982; Lin et al. , 2023). The MYB family constitutes a diverse array of regulatory proteins playing pivotal roles in growth, development, metabolism, and stress responses across a wide range of plant species (Li, B et al. , 2019; Cao et al. , 2020). For example, the R2R3-MYB proteins in Arabidopsis thaliana are involved in drought resistance and disease resistance (Dubos et al. , 2010), with AtMYB15 underpinning drought and cold tolerance in Arabidopsis (Agarwal et al. , 2006). The defining characteristic of MYB transcription factors is the presence of one or more MYB-DNA-binding domains (Dubos et al. , 2010). The MYB domain consists of four imperfectly repeated sequences—R1, R2, R3, and R4—which form a helix-turn-helix structure. The R2 and R3 repeats form the core DNA-binding motif, while other repeats contribute to overall stability and possess additional regulatory elements (So et al. , 2024). MYB transcription factors are key regulators of stress response genes under abiotic stress conditions such as drought, salinity, and high temperature (Li, J et al. , 2019). According to adaptability to salt, plants are classed as either salt-tolerant or salt-sensitive. Halogeton glomeratus are plant communities that thrive in high-salinity and arid regions due to their ability to tolerate harsh salt and drought stress conditions (Yao et al. , 2021). The modified lipid composition of the tonoplast in halophytes contributes to reduced sodium ion leakage into the cytosol, a significant contribution to the salt tolerance observed in these plants (Leach et al. , 1990; Glenn et al. , 1999). Investigating stress-resistant genes in extremophilic halophytes to understand their mechanisms of stress resistance is key to the breeding of new crops with salt and drought tolerance (Huang et al. , 2024). In the present study, we identified and successfully cloned the stress resistance gene HgMYB60 from the full-length transcriptome data of H. glomeratus. Comprehensive analyses of HgMYB60 to the salinity and drought resistance were performed through its overexpression in Arabidopsis and yeast cells. Our research is important for further understanding the regulatory mechanism of HgMYB60 in endowing stress tolerance. 2 Results 2.1 The results of HgMYB60 bioinformatics analysis To investigate the relationships between HgMYBs and AtMYBs, 125 MYBproteins, comprising 58 H. glomeratus MYBs (HgMYBs) and 67 Arabidopsis MYBs (AtMYBs), were used to design a phylogenetic tree. All MYBs were classified into six main groups (Figure 1A), and the phylogenetic tree was divided into six groups according to all MYB gene families. Among them, HgMYB60 was located in the sixth group with 26 MYB genes, including 21 H. glomeratus MYB genes and 5 Arabidopsis MYB genes. The phylogenetic tree showed that HgMYB60 is closely related to HgMYB58, HgMYB58, and HgMYB120 in halophytes and to AtMYB-55 and AtMYB-56 in Arabidopsis . Analysis of the protein physicochemical properties of the 58 MYB family members of H. glomeratus (Table S2) revealed that the number of amino acids ranges from 157 to 1324, with HgMYB129 having the fewest amino acids and HgMYB71 having the most. Those genes encoding proteins that exhibit relatively good stability in vitro, such as HgMYB15 , HgMYB108 , HgMYB129 , HgMYB81 , and HgMYB89 . All of the proteins are hydrophilic. The MYB families of the 58 halophytes are divided into six groups. Motifs 1 and 2 are mainly present in the first branch, whereas motifs 7 and 15 are mainly found in the fourth branch (Figure 1B). The exons and introns of the HgMYB gene family were analyzed to further understand the gene structure of this gene family. In the HgMYB gene family, the number of exons ranges from 3 to 23 (Figure 1C). Specifically, HgMYB91 and HgMYB66 each contain a single exon, while HgMYB114 and HgMYB117 each have two exons. Ten genes possess three exons and 31 genes exhibit between four and nine exons. Additionally, 12 genes contain more than 10 exons. HgMYB genes contain between 1 and 24 introns. Among these genes, four— HgMYB38 , HgMYB48 , HgMYB57 , and HgMYB58 —contain a single intron, nine contain two introns, two— HgMYB7 and HgMYB52 —contain three introns, 33 contain between four and nine introns, and six contain between 11 and 14 introns. HgMYB43 comprises 17 introns, HgMYB44 comprises 19 introns, HgMYB14 comprises 22 introns, and HgMYB39 comprises 24 introns. Most genes have upstream and downstream UTR regions, but some genes lack UTR regions, suchas HgMYB122 , HgMYB81 , HgMYB71 , HgMYB94 , and HgMYB106 . The prediction results of conserved domains indicated that most HgMYB genes contain conserved domains related to MYB transcription factor proteins (Figure 1D). 2.2 Expression patterns of the MYB gene family under salt and drought stress The expression patterns of the AtMYB gene under different salt stress and drought stress conditions were analyzed by using qRT-PCR. The expression levels of most AtMYB genes increased to better manage the damage caused by abiotic stress (Figure 1A, Figure 1B). This suggests that the MYB family plays an important role in organisms subjected to salt and drought stress. 2.3 Subcellular localization of HgMYB60 protein Transient expression of 35S:: HgMYB60 -GFP in Arabidopsis protoplasts revealed its subcellular localization. HgMYB60 -GFP fluorescence was only evident in the nucleus (Figure 2A, Figure 2B), indicating that the HgMYB60 gene is localized to the nucleus, it was consistent with the bioinformatics description. 2.4 Histochemical staining and qRT-PCR analysis in transgenic lines The leaves, flower buds, roots, and lateral roots of transgenic Arabidopsis seedlings containing the upstream promoter of HgMYB60 and WT were stained.The entire plant, leaf veins, flower buds, root tips, vascular tissues of the roots, and lateral roots of the transgenic HgMYB60 lines were all stained with GUS, and no staining was seen in WT (Figure 2C), which indicates that HgMYB60 has some constitutive promoter activity. qRT-PCR analysis showed that the expression of HgMYB60 in increased with the prolong of salt and drought treatments. However, the expression level of HgMYB60 reached highest when the seedlings were treated with 200 mM NaCl for 24 h, and it was highest for transgenic Arabidopsis seedlings under drought stress for 12 h, reaching 25.15-fold compared with the control (Figure 2D). 2.5 HgMYB60 regulates Na + and K + transport in yeast To assess the transport capabilities of Na + and K + for HgMYB60 , the HgMYB60 was introduced into the yeast strains AXT3 and CY162. There was no significant difference in the growth of colonies between the EV strain and the HgMYB60 strain without NaCl (Figure 2E). Lower Na + levels were observed in the medium of HgMYB60 -expressing yeast at 10 and 50 mM Na + concentrations compared to EV-expressing yeast (P < 0.05, Figure 2F). When the K + concentration in the medium was less than 7 mM (0, 0.2 mM, and 1 mM), the yeast strains transformed with EV did not grow. However, when the K + concentration in the medium was 0.2 mM and 1 mM, the yeast strains transformed with HgMYB60 could grow, and they could still grow even when the culture medium was diluted 10 times. When the concentration of K + in the medium reached100 mM, the strains transformed with HgMYB60 grew normally (Figure 2G). At the same time, the K + content showed higher in the HgMYB60 strain than that in EV at 0.2 and 100 mM K + concentration levels (P < 0.05, Figure 2H). This indicates that the HgMYB60 gene can midiates Na + efflux and K + absorption in cell level. 2.6 The salt and drought tolerance of HgMYB60 in Arabidopsis Compared with the control, the seed germination and seedlings growth of transgenic Arabidopsis decreased under salt and drought stress, but which showed higher than that of WT treated with salt and drought stress (Figure 3A) . The seed germination of WT reached 79.44% and 69.16% when it exposed to 100 mM and 200 mM NaCl stress, while those of transgenic seeds were 91.39% and 89.16%, respectively. Under 10% and 20% PEG-6000 stress, the seed germination of WT seeds were 91.94% and 70.83%, while those of transgenic seeds were 97.50% and 93.89%, respectively (Figure 3A). The seedlings growth of transgenic Arabidopsis was not significantly different from that of WT in normal 1/2 MS medium, and which was significantly better than that of WT under 100 mM and 200 mM NaCl stress. The WT seedlings grew slowly under 10% and 20% PEG-6000 treatments, but transgenic Arabidopsis grew normally (Figure 3B). In addition, salt and drought stress inhibited root growth, the root length of transgenic plants was significantly longer than that of WT when it exposed to 100 mM and 200 mM NaCl stress. Seedlings growth of WT was significantly inhibited in the presence of 200 mM NaCl stress, but transgenic plants grew significantly better than WT. The root length of transgenic plants was significantly longer than that of WT when they were exposed to 10% PEG-6000 and 20% PEG-6000 stress (Figure 3C, Figure 3D).The leaves were wilting in WT treated with 200 mM NaCl for 9 d, and which showed well in transgenic Arabidopsis (Figure 3E). Similarly, the WT leaves became wilting when the samples under drought treatment for 9 d, and the transgenic Arabidopsis could still grow normally ( Figure 3E ). Under normal conditions, there was no significant difference in the fresh weight between WT and transgenic Arabidopsis . However, the fresh weights of transgenic Arabidopsis was 1.37- and 3.15 -fold than that of WT when seedlings exposed to salt and natural drought stress for 9 d(Figure 3F). 2.7 The physiological analysis of Arabidopsis under salt and drought stress To further explore the ability of HgMYB60 to confer tolerance to salt and drought stress, the physiological indices of samples treated with salt and drought stress were measured. The activities of SOD, POD, CAT, Pro content and SP content increased first, and then decreased with the prolonging of salt and drought stress, which showed highest for plants treated with salt and drought stress for 6d, and the MDA content increased (Figure 4A-F). Meanwhile, the activities of SOD, POD, CAT and proline content were higher in transgenic Arabidopsis than that of WT under salt and drought stress (Figure 4A-F). Na + content of transgenic Arabidopsis decreased with the prolonging the salt treatments, and it showed lower in transgenic Arabidopsis than that of WT (Figure 4G). Conversely, the K + content of transgenic Arabidopsis was higher than that of WT at 200 mM NaCl for 0, 6, 12, 24 and 48 h (Figure 4H). The K + /Na + ratio at different time points indicated that the WT plants exhibited a lower ratio than the transgenic plants (Figure 4I). Moreover, the K + /Na + ratio of transgenic Arabidopsis reached its maximum value at 24 h. 3 Discussion Soil salinization, drought, and other abiotic stressors have emerged as increasingly critical global challenges that significantly restrict the sustainable development of agriculture worldwide (Ganapati et al. , 2022; Muthuvel et al. , 2023). Therefore, there is an urgent need to identify and characterize novel genes from extreme halophytes in order to boost the drought and salt tolerance of crops. The MYB gene family is one of the largest families of transcription factors in plants. Compared to animals and yeast, the structure and function of MYB proteins are highly conserved in plants. Based on the number of conserved MYB domains, the MYB family can be classified into four subfamilies: MYB-related (1R-MYB), R2R3-MYB (2R-MYB), R1R2R3-MYB (3R-MYB), and 4R-MYB (Du et al. , 2015). In plants, the first identified MYB gene was COLORED1 (C1), which plays a crucial role in anthocyanin biosynthesis in maize kernels (Paz Ares et al. , 1987). Members of the MYB family have been identified and characterized in more than 75 plant species (Wu et al. , 2022), such as Arabidopsis (Stracke et al. , 2001), Oryza sativa (Katiyar et al. , 2012), Brachypodium distachyon (Chen et al. , 2017), and Setaria italica (Muthamilarasan et al. , 2014). Members of the MYB-R2R3 subfamily have been frequently implicated in plant development and stress tolerance (Wu et al. , 2022). In the present study, fifty-eight members of the MYB gene family were identified from the full-length transcriptome data of H. glomeratus . These members encode an average of 548 amino acids, with a mean protein molecular weight of 60.810 KDa, a GRAVY score of −0.6913, and an instability index of 50.78. All identified proteins exhibit hydrophilic properties. Excessive accumulation of Na + within plant tissues can cause significant physiological damage and may ultimately lead to plant death (Munns & Tester, 2008) . Therefore, plants must maintain an optimal concentration of Na + to mitigate the adverse effects of its accumulation. Additionally, the energy conserved by reducing salt stress responses can be redirected toward plant growth (Munns & Gilliham, 2015) . Accordingly, maintaining the intracellular balance of Na + and K + is crucial for enhancing plant salt tolerance (Deinlein et al. , 2014). Previous studies have demonstrated that the transformation of defective yeast strains can effectively illustrate the capability of genes to transport sodium and potassium ions (Al-Harrasi et al. , 2020). In the AXT3 yeast mutant strain, we found that overexpression of HgMYB60 enhanced yeast growth and promoted Na + uptake. Similarly, in the CY162 yeast mutant strain, HgMYB60 overexpression improved yeast growth while facilitating K + absorption. These findings demonstrate that HgMYB60 is capable of participating in Na + efflux transport and enhancing K + absorption. The reduction in cytoplasmic Na + and retention of K + represent one of the key mechanisms in plant salt tolerance. This process can be achieved by removing excess Na + from the cytoplasm via SOS1 or Na + /H + antiporters in the plasma membrane and by sequestering surplus Na + in the vacuole through NHX or vacuolar Na + /H + antiporters (Blumwald & Poole, 1985; Bassil et al. , 2012). Overexpression of AlNHX1 in transgenic soybeans results in decreased Na + levels in the shoots and increased K + concentrations in both the roots and shoots, indicating that NHX1 plays a significant role in maintaining K + homeostasis rather than in sequestering excess Na + (Kobayashi et al. , 2012; Liu et al. , 2014). In the present study, after stable expression of HgMYB60 in Arabidopsis , WT leaves accumulated significantly more Na + over time compared to HgMYB60 -overexpressing Arabidopsis leaves when exposed to salt stress conditions. Conversely, HgMYB60 -overexpressing Arabidopsis leaves accumulated less K + than WT leaves. These findings indicate that HgMYB60 plays a role in Na + efflux and promotes K + uptake in Arabidopsis leaves, consistent with the results from the yeast complementation analysis. K + plays a critical role in numerous enzymatic reactions and is essential for maintaining ion homeostasis and pH stability as a vital macronutrient in plants (Ahmad & Maathuis, 2014). Experimental data indicate that maintaining a high K + /Na + ratio can mitigate the detrimental effects of salt stress on plants (Jbir-Koubaa et al. , 2015). Here, the K + /Na + ratio of transgenic A. thaliana was significantly higher than that of WT with prolonged salt stress. Plants have developed ROS clearance systems as defense mechanisms to combat salt and drought stress. They have also developed antioxidant defense systems, including SOD, POD, CAT, and APX, to mitigate oxidative damage under adverse conditions (Mittler, 2002; Foyer & Noctor, 2005). The tolerance or sensitivity of plants to salt and drought stress is closely linked to their inherent antioxidant response. Plants with higher resistance exhibit greater adaptability and protect themselves from adverse conditions by increasing the activity of antioxidant enzymes (Türkan et al. , 2005; Hediye Sekmen et al. , 2007). The important compatible solutes, such as Pro, SP, and SS, include Pro, which also acts as an antioxidant (Li et al. , 2018). MDA is a product of lipid peroxidation in membrane systems. In the present study, the activities of SOD, POD, and CAT were higher in transgenic Arabidopsis leaves than in WT. Similarly, the Pro content was also higher compared to WT, whereas the MDA and SP contents were lower. Thus, the overexpression of HgMYB60 in Arabidopsis enhanced its salt tolerance. Notably, HgMYB60 was also involved in drought stress regulation. Under drought stress, the activities of SOD, POD, and CAT, as well as Pro content, were significantly higher than those in WT plants, while MDA and SP contents were lower. The physiological responses to salt stress and drought stress exhibited a consistent pattern. Genes are differentially expressed across various growth stages and tissues during plant growth and development. The promoters of these genes play a crucial role in regulating their expression. Promoters are DNA sequence elements located upstream of genes that are recognized and bound by RNA polymerase to initiate transcription. In transgenic technology, the precise regulation of target gene promoters is of paramount importance (Bao et al. , 2024). The leaf veins, flower buds, root tips, and root vascular tubes of Arabidopsis were stained after GUS staining of transgenic Arabidopsis and WT plants expressing HgMYB60 . The promoter of HgMYB60 was able to drive the expression of the GUS gene in different tissues, demonstrating that HgMYB60 functions as an active tissue-specific promoter. qRT-PCR analysis of transgenic Arabidopsis thaliana leaves further confirmed that HgMYB60 positively regulates responses to salt and drought stress. Under both stress conditions, HgMYB60 expression levels gradually increased over time, enabling the HgMYB60 -overexpressing Arabidopsis plants to better tolerate these adverse environmental conditions. In conclusion, HgMYB60 plays a significant role in enhancing salt tolerance and drought resistance in plants . HgMYB60 regulates ion homeostasis in plant cells by enhancing Na + uptake and reducing K + efflux in leaf tissues . This gene enhances the activity of relevant antioxidant enzymes, such as SOD, POD, and CAT, under salt and drought stress, reduces the levels of MDA (malondialdehyde), and fortifies the plant antioxidant defense system. Our results indicate that HgMYB60 is a promising subject for studying the underlying functional mechanisms and for breeding salt-tolerant and drought-resistant varieties. 4 Materials and Methods 4.1 Plant materials and cultivation techniques H. glomeratus was sown in flowerpots with a 1:1 ratio of sand and vermiculite and cultivated in a greenhouse under natural light conditions. A nutrient soil was thoroughly mixed with imidacloprid solution and placed into 100-ml pots. Twelve flowerpots were randomly placed in a rectangular tray and watered with 3 L of water. The next day, we evenly sowed the seeds of wild-type (WT) and T2 generation transgenic Arabidopsis thaliana with HgMYB60 that had been vernalized at 4℃ for 2 days in the pots. The plants were cultivated in a 25℃ growth chamber with 16 h of light and 8 h of darkness. When the Arabidopsis reached the two-leaf stage, thinning was carried out. An appropriate amount of water and nutrient solution was administered every 2 to 3 days. After about 30 days of growth, the transgenic Arabidopsis was screened for resistance. After 45 days of growth, the plants were subjected to 0 and 200 mM NaCl stress treatments, as well as natural drought stress. 4.2 Molecular cloning of HgMYB60 Complete RNA was extracted from the roots of H. glomeratus after 7 days treatment with 100 mM NaCl. The RNA was treated with DNase I and used for cDNA synthesis. The cDNA of HgMYB60 was amplified through PCR using the primer pairs HgMYB60 -F1 and HgMYB60 -R1 (Table S1). The PCR output obtained was cloned into the pM19-T vector, and the DNA sequence was verified by sequencing. Subsequently, this DNA was used to transfer HgMYB60 into various expression vectors for functional studies. 4.3 Bioinformatics analysis of HgMYB60 Using the H. glomeratus whole-transcriptome database (BioProject ID: PRJNA388267), a BLASTp search against the model plant Arabidopsis yielded protein sequences for members of the H. glomeratus MYB gene family. The Arabidopsis MYB gene family protein sequences were procured from the Arabidopsis Information Resource (TAIR) database (https://www.arabidopsis.org/). The amino acid count, isoelectric point, molecular weight, total number of positively and negatively charged residues, aliphatic index, instability index (II), and grand average of hydropathicity (GRAVY) of the encoded proteins were analyzed using the ExPASy database (http://web.expasy.org/protparam/). Hydrophobicity, signal peptide, and subcellular localization predictions were conducted using ProtScale (https://web.expasy.org/protscale/),SignalP4.1(https://services.healthtech.dtu.dk/service.php/), and WoLF PSORT (https://wolfpsort.hgc.jp/). Multiple sequence alignment of MYB protein sequences of H. glomeratus and Arabidopsis was performed using Muscle in MEGA 11. The alignment was then trimmed using trimAL Wrapper in TBtools. Phylogenetic trees were constructed in MEGA 11 using the maximum likelihood estimation (Felsenstein, 1981), with the LG+G and WAG+G models selected as optimal, and a bootstrap method of 1000 replicates. Motif analysis was performed using Multiple Em for Motif Elicitation (MEME) (http://meme-suite.org/tools/meme). Promoter cis-acting element prediction analysis was performed using PlantCARE software (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/) and visualized using the Basic Biosequence View function within TBtools software. 4.4 Analysis of the expression patterns of the MYB family under stresses Arabidopsis Heat Tree Viewer was used to download the gene expression of 66 Arabidopsis genes in the MYB family under different salt and drought conditions, and the expression map was constructed with TBtools. 4.5 Subcellular localization of HgMYB60 The vector pRI101-GFP was linearized using SalI restriction enzyme and the digestion products were purified and recombined with the PCR products using the ClonExpress-II One Step Cloning Kit (Vazyme) with the target gene fragment. The recombinant plasmids were transformed into competent Escherichia coli DH5α cells. The PCR-positive transformants were selected for shake flask culture and plasmid extraction, and the amplification products were sequenced. Leaves of Arabidopsis seedlings at the 5–6 leaf stage were collected and immersed in an appropriate amount of enzymatic solution to treat the tissues (all solution formulations are detailed in Table S3). After filtration, the supernatant was removed by centrifugation, and the precipitate was cleaned twice with a pre-cooled solution. Microscopic examination was performed after 500 μL MMG solution was added to every 200 μL protoplasm for suspension to ensure that 20–40 cells could be seen in each field of vision under 40× magnification. Next, 200 μL protoplast suspension, 10 μL carrier plasmid DNA, and PEG-6000 solution equal to the sum of the volume of DNA and protoplast were evenly mixed and incubated for 30 min at room temperature. Next, 1 mL W5 solution was added to terminate the reaction, and the protoplasts were collected by centrifugation. Then, 1 mL W5 solution was added again and the tissue samples were cleaned twice. After the third addition, the samples were cultured at 28℃ for 48 h away from light. An FV1000 laser confocal microscope was used for observation and imaging. Empty vector expressing GFP gene was used as control and the above steps were followed. 4.6 GUS histochemical analysis and qRT-PCR analysis of HgMYB60 Arabidopsis seeds with HgMYB60 gene were disinfected and evenly planted on 1/2 MS medium. WT Arabidopsis was used as the control, and staining was performed with a GUS staining kit (SL7160 Coolaber Technology Beijing China). Transgenic and WT Arabidopsis plants, leaves, and roots were respectively loaded into 5-mL EP tubes. Then, the appropriate amount of GUS staining solution was added at 37℃ for 12–16 h in the dark, followed by 3 to 4 washes in 70% ethanol to remove chlorophyll. The treated material was photographed under a microscope. Under 200 mM NaCl stress and natural drought treatment for 0, 6, 12, 24, and 48 h, the leaves of Arabidopsis HgMYB60 were cleaned and disinfected and quickly stored in liquid nitrogen. RNA from Arabidopsis leaves was extracted using an RNA Simple Total RNA Kit (DP419 Tiangen Biotech Beijing, China). cDNA was reverse-transcribed using a Tiangen reverse transcription kit (KR118-02 Tiangen Biotech Beijing, China). NCBI Prime-BLAST was used to design qRT-PCR primers, with AtActin as the internal reference gene (Table S1). The total reaction volume was 20 μL, comprising 10 μL 2×SuperReal PreMix Plus, 0.6 μL positive and reverse primer (10 μmol/L), 1 μL cDNA template (100 ng/μL), and 7.8 μL RNase-free ddH 2 O. The reaction procedure involved two steps: predenaturation at 95℃ for 15 min, followed by 40 cycles of denaturation at 95℃ for 10 s and annealing at 60℃ for 32 s. The relative expression of genes was calculated using the 2 -ΔΔCT formula (Livak & Schmittgen, 2001). Three technical replicates were obtained from each analysis. 4.7 Heterologous expression of HgMYB60 in yeast HgMYB60 CDNA was cloned into yeast plasmid empty vector (EV) (Fig. S1). Recombinant vector (EV- HgMYB60 ) or EV were introduced into Na + -sensitive yeast ( S. cerevisiae ) strain AXT3 (Amtmann et al. , 2001) and K + absorption-deficient yeast strain CY162 (Mäser et al. , 2002). The ability of the HgMYB60 yeast strain to transport Na + and K + was tested on AP solid medium with different concentrations of Na + and K + using the yeast spot method and compared with that of EV (Patankar et al. , 2018). The behavior of HgMYB60 and EV yeast cells was verified in AP liquid medium containing different concentrations of Na + and K + (Patankar et al. , 2019). Yeast colonies were used to evaluate the effects of HgMYB60 and EV yeast cells on the salinity response. 4.8 Salt and drought tolerance of transgenic Arabidopsis Each Arabidopsis plant with the HgMYB60 gene was labeled, and leaves of about 2–5 mm 2 were taken from each plant to extract DNA. After the PCR, detection was performed using 1% agarose gel electrophoresis, and the gel was visualized using a gel electrophoresis imaging instrument (Fig. S2).The false-positive plants were removed for further analysis. Arabidopsis seeds were washed with 1% NaClO for 10 min, followed by 70% ethanol for 1 min and 4–5 washes in ddH 2 O. They were then unifo rmly seeded on 1/2 MS medium (0, 100, and 200 mM, 10% and 20% PEG-6000) with three biological replicates per treatment. The seeds were cultured in an incubator under a 16 h/8 h light/dark cycle at a temperature of 22℃ and humidity of 70%. The germination rate was calculated after 7 days. Root length was measured when the Arabidopsis plants reached 20 days. 4.9 Physiological analysis of transgenic Arabidopsis SOD activity was determined using the nitroblue tetrazolium (NBT) photoreduction method (Fu & Huang, 2001). POD activity was measured using the guaiacol method (Xing et al. , 2010). CAT activity was determined using the ultraviolet absorption method (Cakmak & Marschner, 1992). MDA content was measured using the thiobarbituric acid method (Zhang et al. , 2008). Soluble protein (SP) content was determined using the Coomassie Brilliant Blue G-250 staining method (Liu et al. , 2019). Proline (Pro) content was calculated using the acid ninhydrin colorimetric method (Redillas et al. , 2012). 4.10 The content of Na + and K + in transgenic Arabidopsis Two kinds of Arabidopsis leaves were collected under 200 mM NaCl stress and natural drought treatment for 0, 6, 12, 24, and 48 h, dried to constant weight at 65℃, and crushed into a 0.3-mm sieve. Then, to 0.1 g of the sample, 5 mL of concentrated H 2 SO 4 was added, mixed well, and boiled at 400℃. When the boiling liquid turned brown, it was removed and let cool slightly. Then, hydrogen peroxide was added drop by drop with shaking of the digestion tube to accelerate the reaction until the boiling liquid became colorless or clarified. The solution was boiled for another 10 min, cooled to room temperature, and transferred to a 100-mL volumetric bottle for determination of Na + and K + content. Three biological replicates were obtained. 4.11 Statistical analysis Statistical evaluations were performed using IBM SPSS Statistics version 21. Data analysis and mapping was conducted using Origin 2022. Bioinformatics analysis and visualization was performed TBtools and MEGA11. One-way analysis of variance (ANOVA) was used to evaluate the differences among means. Duncan’s new multiple range test was used to determine the significance of variance among the means of the examined variables. A p-value < 0.05 was considered significant. Acknowledgements Fuxi Talent Project of Gansu Agricultural University (GAUfx-04Y011; Gaufx- 03Y06);Provincial Youth Science and Technology Fund of Gansu Province (22JR5RA880); Special Projects for East West Science and Technology Cooperation (25CXNA030);National Natural Science Foundation of China (Grant 31960072; 32001514); Industrial Support Project of Colleges and Universities in Gansu Province (Grant 2021CYZC-12); the Earmarked Fund for China Agriculture Research System (CARS-05-02A-02); Innovation Capacity Enhancement Project of Gansu Education Department (Grant 2019A-053). Conflicts of Interest The authors declare that they have no conflict of interest. Data Availability Statement The data that support the findings of this study are available from the corresponding author upon reasonable request. Author contributions H.W. and J.W.designed the experiments;X.S.,S.L.,Z.Z., Y.M., E.S., B.L. and X.M. conducted the experiments; P.H. wrote the manuscript; L.R., H.Z.,P.H., K.Y. and Y.Y. analyzed the data; All authors have read and agreed to the published version of the manuscript. Supporting Information Fig. 1. (A) Phylogenetic analysis of MYB. Different colors indicate the genes of different species. Halogeton is in blue while Arabidopsis is in green. (B) Conserved motifs in Halogeton MYB protein. (C) Halogeton MYB gene structure. (D) Conserved motifs of Halogeton MYB proteins. (E) Model map of Arabidopsis MYB gene expression under different salt stress conditions. The horizontal axis shows different tissues and different stress treatments.(F) Model map of Arabidopsis MYB gene expression under different drought stress conditions. The horizontal axis shows different tissues and different stress treatments. Fig. 2. (A) Localization of HgMYB60 -GFP in Arabidopsis protoplasts. Images from left to right show HgMYB60 -GFP fluorescence, chloroplast fluorescence, a bright-field image, and a merge of the three images. Scale bar = 10 μm. (B) Localization of free GFP in Arabidopsis protoplasts. Scale bar = 10 μm. (C) GUS staining of HgMYB60 in different tissues of Arabidopsis . Scale bar = 500 px. (D) qRT-PCR expression in leaves of HgMYB60 -overexpressing plants under salt and drought stress. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05. (E) The yeast strain AXT3 transformed with HgMYB60 or EV was grown on AP solid medium with the designated concentrations of Na + to assay the response of the transgenic yeast. (F) The concentration of Na + in transgenic HgMYB60 and EV yeast cells after 3-day culture in AP liquid medium with the specified Na + concentration. (G) The yeast strain CY162 transformed with HgMYB60 or EV was grown on AP solid medium with the designated concentrations of K + to assay the response of the transgenic yeast. (H) The concentration of K + in transgenic HgMYB60 and EV yeast cells after 3-day culture in AP liquid medium with the specified K + concentration. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates, each being an independent colony. Two technical replicates were averaged). Fig. 3. (A) Seed germination phenotypes of wild-type and transgenic strains on MS medium supplied with 0, 100, 200 mM NaCl and 10%,20% PEG-6000 for 7 days. (B) Seed germination of wild-type and transgenic strains on MS medium containing 0, 100, and 200 mM NaCl and 10% and 20% PEG-6000 for 7 days. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). (C) WT and HgSMYB60 strains were observed after 20 days of growth on 1/2 MS medium with different concentrations of NaCl (100 and 200 mM) and different concentrations of PEG-6000 (10% and 20%). Scale bar = 10 mm. (D) Root length of WT and HgSMYB60 plants was determined after 20 days of growth on 1/2 MS medium containing different concentrations of NaCl (100 and 200 mM) and different concentrations of PEG-6000 (10% and 20%). The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). (E) The performance of genetically modified HgMYB60 and wild-type plants growing in soil after exposure to natural drought and salinity (200 mM NaCl) for 3, 6, and 9 days. (F) Fresh weight of transgenic Arabidopsis thaliana on day 9 of natural drought and salinity (200 mM NaCl). The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). Fig. 3. (A) Superoxide dismutase (SOD) activity in leaves of plants overexpressing HgMYB60 under salt stress and drought stress for 0,3,6 and 9 days (B) Peroxidase (POD) Activity in Leaves of Plants Overexpressing HgMYB60 under Salt Stress and Drought Stress for 0, 3, 6 and 9 Days. (C) Catalase (CAT) Activity in Leaves of Plants Overexpressing HgMYB60 under Salt Stress and Drought Stress for 0,3,6 and 9 Days. (D) Malondialdehyde (MDA) Content of Leaves of Plants Overexpressing HgMYB60 under Salt Stress and Drought Stress for 0,3,6 and 9 Days. (E) Proline (Pro) Content of Leaves of Plants Overexpressing HgMYB60 under Salt Stress and Drought Stress for 0,3,6 and 9 Days. (F) Soluble Protein (SP) Content of Leaves of Plants Overexpressing HgMYB60 under Salt Stress and Drought Stress for 0,3,6 and 9 Days. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). (G) Effect of salt stress on the content of sodium ion in plants overexpressing HgMYB60 . Transgenic HgMYB60 and wild-type plants growing in soil and subjected to natural drought and salt stress (200 mM NaCl) for 3, 6, and 9 day were analyzed to determine the levels of physiologically active substances in leaves. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). (H) Effect of salt stress on the content of potassium ion in plants overexpressing HgMYB60 . Transgenic HgMYB60 and wild-type plants growing in soil and subjected to natural drought and salt stress (200 mM NaCl) for 3, 6, and 9 day were analyzed to determine the levels of physiologically active substances in leaves. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). (I) Effect of salt stress on the content of Sodium-potassium ion ratio in plants overexpressing HgMYB60 . Transgenic HgMYB60 and wild-type plants growing in soil and subjected to natural drought and salt stress (200 mM NaCl) for 3, 6, and 9 day were analyzed to determine the levels of physiologically active substances in leaves. The different letters denote significant differences by Duncan’s new multiple range test at P < 0.05 (data are the mean ± SE of n = 3 biological replicates). Supplementary Material Fig. S1. Amplification of the CDS sequence of HgMYB60 in H. glomeratus . PCR product. Fig. S2. PCR identification of positive overexpressing plants. M: DL15000+2000 DNA Marker. 1–4: resistant seedling number. 5–6: False-positive seedling number. +: recombinant plasmid control. -: pure water blank control. Table S1. Primers used in this study. Table S2. Physicochemical properties of MYB family proteins of H. glomeratus . Table S3. Reagent preparation. References Agarwal, M., Hao, Y. Kapoor, A. Dong, C. et al. 2006. A R2R3 type MYB transcription factor is involved in the cold regulation of CBF genes and in acquired freezing tolerance. Journal of Biological Chemistry 281(49): 37636-37645. https://doi.org/10.1074/jbc.M605895200. Ahmad, I., Maathuis, FJ. 2014. Cellular and tissue distribution of potassium: physiological relevance, mechanisms and regulation. Journal of Plant Physiology 171(9): 708-714. https://doi.org/ 10.1016/j.jplph.2013.10.016. Al-Harrasi, I., Jana, GA. Patankar, HV. Al-Yahyai, R. et al. 2020. A novel tonoplast Na + /H + antiporter gene from date palm (PdNHX6) confers enhanced salt tolerance response in Arabidopsis. Plant Cell Reports 39: 1079-1093. https://doi.org/ 10.1007/s00299-020-02549-5. Amtmann, A., Fischer, M. Marsh, EL. Stefanovic, A. et al.2001. The wheat cDNA LCT1 generates hypersensitivity to sodium in a salt-sensitive yeast strain. Plant Physiology 126(3): 1061-1071. https://doi.org/10.1104/pp.126.3.1061. Angon, PB., Tahjib-Ul-Arif, M. Samin, SI. Habiba, U. et al.2022. How do plants respond to combined drought and salinity stress?—a systematic review. Plants Basel 11(21): 2884. https://doi.org/ 10.1016/j.jplph.2005.05.003. Bao, A., Jiao, T. Hu, T. Cui, K. Yue, W. et al. 2024. Cloning of the Arabidopsis SMAP2 promoter and analysis of its expression activity. Scientific Reports 14(1): 11451. https://doi.org/10.1038/s41598-024-61525-1. Bassil, E., Coku, A. Blumwald, E. 2012. Cellular ion homeostasis: emerging roles of intracellular NHX Na + /H + antiporters in plant growth and development. Journal of Experimental Botany 63(16): 5727-5740. https://doi.org/10.1093/jxb/ers250. Blumwald, E., Poole, RJ. 1985. Na + /H + antiport in isolated tonoplast vesicles from storage tissue of Beta vulgaris. Plant Physiology 78(1): 163-167. https://doi.org/10.1104/pp.78.1.163. Cakmak, I., Marschner, H. 1992. Magnesium deficiency and high light intensity enhance activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase in bean leaves. Plant Physiology 98(4): 1222-1227. https://doi.org/10.1104/pp.98.4.1222. Cao, Y., Li, K. Li, Y. Zhao, X. Wang, L. 2020. MYB transcription factors as regulators of secondary metabolism in plants. Biology 9(3): 61. https://doi.org/10.3390/biology9030061. Chen, S., Niu, X. Guan, Y. Li, H. 2017. Genome-wide analysis and expression profiles of the MYB genes in Brachypodium distachyon. Plant and Cell Physiology 58(10): https://doi.org/1777-1788. 10.1093/pcp/pcx115. Deinlein, U., Stephan, A,B. Horie, T. Luo, W, Xu. G, Schroeder JI. 2014. Plant salt-tolerance mechanisms. Trends in Plant Science 19(6): 371-379. https://doi.org/10.1016/j.tplants.2014.02.001. Devi, R., Kaur, N. Gupta, A,K. 2012. Potential of antioxidant enzymes in depicting drought tolerance of wheat (Triticum aestivum L.). https://doi.org/10.1016/j.bpc.2012.09.002. Du, H., Liang, Z. Zhao, S. Nan, M. Tran, L,P. et al.2015. The evolutionary history of R2R3-MYB proteins across 50 eukaryotes: new insights into subfamily classification and expansion. Scientific Reports 5(1): https://doi.org/11037. 10.1038/srep11037. Dubos, C., Stracke, R. Grotewold, E. Weisshaar, B. Martin, C. Lepiniec, L. 2010. MYB transcription factors in Arabidopsis. Trends in Plant Science 15(10): 573-581. https://doi.org/. Foyer, C,H., Noctor, G. 2005. Oxidant and antioxidant signalling in plants: a re‐evaluation of the concept of oxidative stress in a physiological context. Plant, Cell and Environment 28(8): 1056-1071. https://doi.org/ 10.1111/j.1365-3040.2005.01327.x. Fu, J., Huang, B. 2001. Involvement of antioxidants and lipid peroxidation in the adaptation of two cool-season grasses to localized drought stress. Environmental and Experimental Botany 45(2): 105-114. https://doi.org/ 10.1016/S0098-8472(00)00084-8. Ganapati, R,K., Naveed, S,A. Zafar, S. Wang, W. Xu, J. 2022. Saline-alkali tolerance in rice: Physiological response, molecular mechanism, and QTL identification and application to breeding. Rice Science 29(5): 412-434. https://doi.org/ 10.1016/j.rsci.2022.05.002. Gill, S,S., Tuteja, N. 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry 48(12): 909-930. https://doi.org/10.1016/j.plaphy.2010.08.016. Glenn, E,P., Brown, J,J. Blumwald, E. 1999. Salt tolerance and crop potential of halophytes. Critical Reviews in Plant Sciences 18(2): 227-255. https://doi.org/ 10.1080/07352689991309207. Hamaji, K., Nagira, M. Yoshida, K. Ohnishi, M. Oda, Y. et al.2009. Dynamic aspects of ion accumulation by vesicle traffic under salt stress in Arabidopsis. Plant and Cell Physiology 50(12): https://doi.org/2023-2033. 10.1093/pcp/pcp143 Hasanuzzaman, M., Bhuyan, M,B. Anee, T,I. et al.2019. Regulation of ascorbate-glutathione pathway in mitigating oxidative damage in plants under abiotic stress. Antioxidants 8(9): 384. https://doi.org/ 10.3390/antiox8090384. Hediye, Sekmen, A., Türkan, İ. Takio, S. 2007. Differential responses of antioxidative enzymes and lipid peroxidation to salt stress in salt‐tolerant Plantago maritima and salt‐sensitive Plantago media. Physiologia Plantarum 131(3): 399-411. https://doi.org/10.1111/j.1399-3054.2007.00970.x. Huang, Z., Yao, L. Li, B. Ma, X. et al. 2024. HgS2, a novel salt‐responsive gene from the Halophyte Halogeton glomeratus, confers salt tolerance in transgenic Arabidopsis. Physiologia Plantarum 176(3): e14356. https://doi.org/ 10.1111/ppl.14356. James, R,A., Blake, C. Byrt, C,S. Munns, R. 2011. Major genes for Na+ exclusion, Nax1 and Nax2 (wheat HKT1; 4 and HKT1; 5), decrease Na + accumulation in bread wheat leaves under saline and waterlogged conditions. Journal of Experimental Botany 62(8): 2939-2947. https://doi.org/ 10.1093/jxb/err003. Jbir-Koubaa, R., Charfeddine, S. Ellouz, W. Saidi, M,N. et al.2015. Investigation of the response to salinity and to oxidative stress of interspecific potato somatic hybrids grown in a greenhouse. Plant Cell, Tissue and Organ Culture 120: 933-947. https://doi.org/10.1007/s11240-014-0648-4. Katiyar, A., Smita, S. Lenka, S,K. Rajwanshi, R. Chinnusamy, V. Bansal, K,C. 2012. Genome-wide classification and expression analysis of MYB transcription factor families in rice and Arabidopsis. BMC Genomics 13: 1-19. https://doi.org/ 10.1186/1471-2164-13-544. Klempnauer, K., Gonda, T,J. Bishop, J,M. 1982. Nucleotide sequence of the retroviral leukemia gene v-myb and its cellular progenitor c-myb: the architecture of a transduced oncogene. Cell 31(2): 453-463. https://doi.org/ 10.1016/0092-8674(82)90138-6. Kobayashi, S., Abe, N. Yoshida, K,T. Liu, S. Takano, T. 2012. Molecular cloning and characterization of plasma membrane-and vacuolar-type Na + /H + antiporters of an alkaline-salt-tolerant monocot, Puccinellia tenuiflora. Journal of Plant Research 125: 587-594. https://doi.org/ 10.1007/s10265-012-0475-9. Kumar, M., Hasan, M. Arora, A. Gaikwad, K. Kumar, S. Rai, R,D. et al. Sodium chloride-induced spatial and temporal manifestation in membrane stability index and protein profiles of contrasting wheat ( Triticum aestivum L.) genotypes under salt stress. Ind. J Plant Physiol. 2015; 20:271-275. 10.1007/s40502-015-0157-4 Leach, R,P., Wheeler, K,P. Flowers, T,J. Yeo, A,R. 1990. Molecular Markers for Ion Compartmentation in Cells of Higher Plants. Journal of Experimental Botany 41(9): https://doi.org/ 1089-1994. 10.1093/JXB/41.9.1079. Li, B., Fan, R. Guo, S. Wang, P. Zhu, X. Fan, Y. Chen, Y. et al. 2019. The Arabidopsis MYB transcription factor, MYB111 modulates salt responses by regulating flavonoid biosynthesis. Environmental and Experimental Botany 166: 103807. https://doi.org/ 10.1016/j.envexpbot.2019.103807. Li, H., Shi, J. Wang, Z. Zhang, W. Yang, H. 2020. H 2 S pretreatment mitigates the alkaline salt stress on Malus hupehensis roots by regulating Na + /K + homeostasis and oxidative stress. Plant Physiology and Biochemistry 156: 233-241. https://doi.org/ 10.1016/j.plaphy.2020.09.009. Li, J., Han, G. Sun, C. Sui, N. 2019. Research advances of MYB transcription factors in plant stress resistance and breeding. Plant Signaling and Behavior 14(8): 1613131. https://doi.org/ 10.1080/15592324.2019.1613131. Li, L., Li, H. Wu, L. Qi, H. 2022. Sulfur dioxide improves drought tolerance through activating Ca 2+ signaling pathways in wheat seedlings. Ecotoxicology 31(5): 852-859. https://doi.org/10.1007/s10646-022-02547-1. Li, W., Qiang, X. Han, X. et al.2018. Ectopic expression of a Thellungiella salsuginea aquaporin gene, TsPIP1; 1, increased the salt tolerance of rice. International Journal of Molecular Sciences 19(8): 2229. https://doi.org/ 10.3390/ijms19082229. Li, X., Li, S. Wang, J. Lin, J. 2020. Exogenous abscisic acid alleviates harmful effect of salt and alkali stresses on wheat seedlings. International Journal of Environmental Research and Public Health 17(11): 3770. https://doi.org/ 10.3390/ijerph17113770. Lin, M., Dong, Z. Zhou, H. Wu, G. Xu, L. et al. 2023. Genome-wide identification and transcriptional analysis of the MYB gene family in pearl millet ( Pennisetum glaucum ). International Journal of Molecular Sciences 24(3): 2484. https://doi.org/ 10.3390/ijms24032484 Liu, H., Li, H. Ning, H. Zhang, X. Li, S. et al.2019. Optimizing irrigation frequency and amount to balance yield, fruit quality and water use efficiency of greenhouse tomato. Agricultural Water Management 226: 105787. https://doi.org/10.1016/j.agwat.2019.105787. Liu, J., Zhang, S. Dong, L. Chu, J. 2014. Incorporation of Na + /H + antiporter gene from Aeluropus littoralis confers salt tolerance in soybean (Glycine max L). Indian Journal of Biochemistry and Biophysics 51.1(2014):58-65. https://doi.org/ 10.1017/S0033583514000018. Livak, K,J., Schmittgen, T,D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 − ΔΔC T method. METHODS 25(4): 402-408. https://doi.org/10.1006/meth.2001.1262 Man WU, Xiangzhu W, Haiyan L, Liyu Y, Qi WU, Cuiping M, Pu S. 2024. Hot Issues and Visual Analysis of Water and Fertilizer Integration Research. Journal of Agriculture 14(4): 52. https://doi.org/10.11923/j.issn.2095-4050.cjas2023-0096 Mäser, P., Eckelman, B. Vaidyanathan, R. et al.2002. Altered shoot/root Na + distribution and bifurcating salt sensitivity in Arabidopsis by genetic disruption of the Na + transporter AtHKT1. Febs Letters 531(2): 157-161. https://doi.org/ 10.1016/S0014-5793(02)03488-9. Mittler, R. 2002. Oxidative stress, antioxidants and stress tolerance. Trends in Plant Science 7(9): 405-410. https://doi.org/ 10.1016/S1360-1385(02)02312-9. Munns, R., Gilliham, M. 2015. Salinity tolerance of crops–what is the cost? New Phytologist 208(3): 668-673. https://doi.org/10.1111/nph.13519. Munns, R., Tester, M. 2008. Mechanisms of salinity tolerance. Annual Review of Plant Biology 59(1): 651-681. https://doi.org/10.1146/annurev.arplant.59.032607.092911. Muthamilarasan, M., Khandelwal, R. Yadav, C,B.et al. 2014. Identification and molecular characterization of MYB transcription factor superfamily in C4 model plant foxtail millet (Setaria italica L.). PLoS One 9(10): e109920. https://doi.org/10.1371/journal.pone.0109920. Muthuvel, D., Sivakumar, B. Mahesha, A. 2023. Future global concurrent droughts and their effects on maize yield. Science of the Total Environment 855: 158860. https://doi.org/10.1016/j.scitotenv.2022.158860 Nadeem, M., Li, J., Yahya, M. Sher, A. Ma, C. Wang, X. Qiu, L. 2019. Research progress and perspective on drought stress in legumes: a review. International Journal of Molecular Sciences 20(10): 2541. https://doi.org/10.3390/ijms20102541. Nardini, A., 2022. Hard and tough: the coordination between leaf mechanical resistance and drought tolerance. Flora 288: 152023. https://doi.org/10.1016/j.flora.2022.152023. Patankar, H,V., Al-Harrasi, I. Al-Yahyai, R. Yaish, M,W. 2018. Identification of candidate genes involved in the salt tolerance of Date Palm (Phoenix dactylifera L.) Based on a Yeast functional bioassay. Dna and Cell Biology 37(6): https://doi.org/524-534. 10.1089/dna.2018.4159. Patankar, S., Yang, S,T. Bayramian, A,J. Bowers, M,W. Datte, P,S. et al.2019. High Intensity 5th Harmonic Generation using CLBO. 2019 Conference on Lasers and Electro-Optics (CLEO). IEEE. https://doi.org/10.1364/cleo_qels.2019.fth1m.7. Paz, Ares, J., Ghosal, D. Wienand, U. Peterson, P,A. Saedler, H. 1987. The regulatory c1 locus of Zea mays encodes a protein with homology to myb proto‐oncogene products and with structural similarities to transcriptional activators. The Embo journal 6(12): 3553-3558. https://doi.org/10.1002/j.1460-2075.1987.tb02684.x. Razi, K., Muneer, S. 2021. Drought stress-induced physiological mechanisms, signaling pathways and molecular response of chloroplasts in common vegetable crops. Critical Reviews in Biotechnology 41(5): 669-691. https://doi.org/10.1080/07388551.2021.1874280. Redillas, M,C., Park, S. Lee, J,W. Kim, Y,S. et al. 2012. Accumulation of trehalose increases soluble sugar contents in rice plants conferring tolerance to drought and salt stress. Plant Biotechnology Reports 6: 89-96. https://doi.org/10.1007/s11816-011-0210-3. Shabala, S. 2013. Learning from halophytes: physiological basis and strategies to improve abiotic stress tolerance in crops. Annals of Botany 112(7): 1209-1221. https://doi.org/10.1093/aob/mct205. Shahzad,H., Ullah, S. Iqbal, M. Bilal, H,M. et al. 2019. Salinity types and level-based effects on the growth, physiology and nutrient contents of maize (Zea mays). Italian Journal of Agronomy 14(4): https://doi.org/199-207. 10.4081/ija.2019.1326. Sharma, A., Shahzad, B. Kumar, V. Kohli, S,K. Sidhu, G,P,S. et al. 2019. Phytohormones regulate accumulation of osmolytes under abiotic stress. Biomolecules 9(7): 285. https://doi.org/10.3390/biom9070285. So, K., Wang, J. Sun, S. Che, H. Zhang, Y. 2024. Comprehensive analysis of MYB gene family and their expression under various stress conditions in Lilium pumilum. Scientia Horticulturae 327: 112764. https://doi.org/10.1016/j.scienta.2023.112764. Stracke, R., Werber, M. Weisshaar, B. 2001. The R2R3-MYB gene family in Arabidopsis thaliana. Current Opinion in Plant Biology 4(5): 447-456. https://doi.org/10.1016/S1369-5266(00)00199-0. Türkan, I., Bor, M. Özdemir, F. Koca, H. 2005. Differential responses of lipid peroxidation and antioxidants in the leaves of drought-tolerant P. Acutifolius Gray and drought-sensitive P. Vulgaris L. Subjected to polyethylene glycol mediated water stress. Plant Science 168(1): 223-231. https://doi.org/10.1016/j.plantsci.2004.07.032. Umair, Hassan, M., Aamer, M. UmerChattha, M. Haiying, T. et al.2020. The Critical Role of Zinc in Plants Facing the Drought Stress. Agriculture , 10, Article,10(9), 396. https://doi.org/10.3390/agriculture10090396. Wu, Y., Wen, J. Xia, Y. Zhang, L. Du, H. 2022. Evolution and functional diversification of R2R3-MYB transcription factors in plants. Horticulture Research , 9(1), 1132-1147. https://doi.org/10.1093/hr/uhac0583. Xing, W., Li, D. Liu, G. 2010. Antioxidative responses of Elodea nuttallii (Planch.) H. St. John to short-term iron exposure. Plant Physiology and Biochemistry 48(10-11): 873-878. https://doi.org/10.1016/j.plaphy.2010.08.006. Yang, X., Lu, M. Wang, Y. et al. 2021. Response mechanism of plants to drought stress. Horticulturae 7(3): 50. https://doi.org/10.19579/j.cnki.plant-d.p.2023.05.009. Yao, L., Wang, J. Li, B. Meng, Y. Ma, X. et al.2021. Influences of heavy metals and salt on seed germination and seedling characteristics of halophyte Halogeton glomeratus. Bulletin of Environmental Contamination and Toxicology 106: 545-556. https://doi.org/10.1007/s00128-021-03130-w. Zhang, S. Hua, B. Zhang, F. 2008. Induction of the activities of antioxidative enzymes and the levels of malondialdehyde in cucumber seedlings as a consequence of Bemisia tabaci (Hemiptera: Aleyrodidae) infestation. Arthropod Plant Interactions 2: 209-213. https://doi.org/10.1007/s11829-008-9044-5. Zhang, S., Wu, Q. Liu, L. Zhang, H. Gao, J. Pei, Z. 2020. Osmotic stress alters circadian cytosolic Ca 2+ oscillations and OSCA1 is required in circadian gated stress adaptation. Plant Signaling and Behavior 15(12): 1836883. https://doi.org/10.1080/15592324.2020.1836883. Zhu, J. 2003. Regulation of ion homeostasis under salt stress. Current Opinion in Plant Biology 6(5): 441-445. https://doi.org/10.1016/S1369-5266(03)00085-2. Supplementary Material File (table.docx) Download 29.67 KB Information & Authors Information Version history V1 Version 1 16 April 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords drought，transport genome myb salt transcription factor transcriptome Authors Affiliations Pengxu He [email protected] Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Lirong Yao Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Xiangling Sun Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Lujuan Sun Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Zhenghuang Zhang Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Shangqing Hu Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Yan Yan Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Baochun Li Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Hong Zhang Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Xiaole Ma Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Erjing Si Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Ke Yang Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Yaxiong Meng Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Huajun Wang Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Juncheng Wang Gansu Agricultural University State Key Laboratory of Aridland Crop Science View all articles by this author Metrics & Citations Metrics Article Usage 189 views 115 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Pengxu He, Lirong Yao, Xiangling Sun, et al. A new transcription factor, HgMYB60, regulates salt and drought stress of halophyte Halogeton glomeratus. Authorea . 16 April 2025. DOI: https://doi.org/10.22541/au.174480048.89154542/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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