Soil alkalinity severely limits the productivity of strawberry, a high-value horticultural crop. The root endophytic fungus Piriformospora indica enhances plant stress resilience, yet the systemic mechanisms underlying its promotion of alkaline tolerance remain poorly understood. This study employed an integrated transcriptomic and metabolomic approach to elucidate these mechanisms in strawberry under alkaline stress. Inoculation with P. indica significantly alleviated stress-induced growth inhibition, improving biomass accumulation and leaf development. Metabolomic profiling identified 1,352 DAMs, predominantly flavonoids and phenolic acids. Transcriptome analysis revealed 19,689 DEGs enriched in oxidoreductase activity, hormone signaling, and secondary metabolism. Multi-omics integration highlighted coordinated changes in the metabolism of pyruvate,alanine, aspartate and glutamate. P. indica maintained the balance of carbon and nitrogen allocation in strawberry under alkaline stress, an effect linked to the downregulation of argG, asnB, and GLT1. These findings suggest a putative systemic metabolic mechanism by which P. indica may enhance alkaline tolerance, potentially through rebalancing primary metabolism. This offers molecular insights into fungal-mediated stress adaptation in strawberry and supports its potential as a sustainable bio-inoculant for improving productivity in alkaline soils.

The first T2T reference genome of Guazuma ulmifolia is reported, which serves as a core genomic resource for stress adaptation research and stress-tolerant breeding in cacao wild relatives. Climate change, particularly increased incidence of drought, poses a major threat to food security. Understanding the genomic basis of environmental adaptation in crop wild relatives can provide valuable resources for improving stress resilience. Guazuma ulmifolia, a wild relative of Theobroma cacao with important ecological and medicinal value, lacks high-quality reference genomic resources. Here, we report the first telomere-to-telomere (T2T) chromosome-level genome assembly of G. ulmifolia, with a genome size of 311.31 Mb, contig N50 of 35.19 Mb, and 98.70

Jojoba (Simmondsia chinensis) is an important woody oil crop, but its cultivation is severely limited by extreme sensitivity to low temperatures. Pathogenesis-related protein 4 (PR4) genes participate in plant stress responses and are frequently associated with jasmonic acid (JA) signaling, but their functions in woody crops remain poorly understood. Here, six class II ScPR4 genes containing conserved Barwin-like domains were identified in jojoba. Phylogenetic and collinearity analyses revealed lineage-specific expansion of the family, mainly driven by tandem duplication. Among these genes, ScPR4–1 contained low-temperature- and MeJA-responsive cis-elements and was strongly induced by both treatments. Functional analyses in Arabidopsis, yeast, and jojoba hairy roots showed that ScPR4–1 enhanced cold tolerance by reducing ROS accumulation and membrane lipid peroxidation and promoting osmotic adjustment. Mechanistic studies further demonstrated that ScMYC2a directly bound to two G-box elements in the ScPR4–1 promoter and activated its transcription. DIECA treatment attenuated ScPR4–1 induction and partially weakened the protective effects of its overexpression, supporting the contribution of JA biosynthesis to this regulation. Together, these findings established a “JA–ScMYC2a–ScPR4–1” regulatory module underlying cold response in jojoba, providing novel insights and candidate targets for cold-tolerance improvement in woody crops.

Scutellaria baicalensis is an important medicinal plant, and the diversity of its rhizosphere microbiota may influence its growth, development, and yield. Numerous studies have reported that warming associated with global climate change significantly altered plant-associated soil microbial diversity. To reveal the effects of night-time warming on the rhizosphere microbial community of S. baicalensis, soil microbial diversity in the rhizosphere (RS) and bulk soil (BS) of S. baicalensis were analyzed by employing bacterial 16S rRNA and fungal ITS sequencing technology. Warming significantly altered both bacterial and fungal communities in the rhizosphere and bulk soils of S. baicalensis, with pronounced changes in OTU composition, relative abundances at both phylum and species levels. The analysis of alpha and beta diversity showed that warming significantly altered the fungal community structure in the rhizosphere soil (R-2 = 0.423, p < 0.05) and significantly reduced the species richness in the bulk soil of S. baicalensis (Shannon and Simpson index, p < 0.05). LEfSe and functional prediction analyses revealed that warming altered the taxonomic composition of both bacterial (35 taxa, LDA > 3) and fungal (24 taxa, LDA > 4) communities in rhizosphere and bulk soils of S. baicalensis, with multiple bacterial and fungal taxa serving as treatment-specific biomarkers. Functional predictions indicated that fungal functional groups, including saprotrophic and mycorrhizal guilds, were more strongly affected by warming than bacteria. Overall, warming has a significantly stronger impact on fungal communities in the rhizosphere and bulk soils of S. baicalensis than on bacteria, and has a significantly greater effect on the diversity of microbial communities in bulk soils than that in rhizosphere soils. This study provides important data for understanding the impact of global climate change on the rhizosphere microbial communities of cultivated plants.

Jojoba [Simmondsia chinensis (Link) Schneider] is sensitive to low temperatures, which hinders its cultivation in temperate arid regions. The apoplast is a cellular component outside the plasma membrane of plant cells and is widely involved in the response of plants to environmental stress. Here, we used the infiltration-centrifugation method to extract apoplast fluid from jojoba leaves and analyzed changes in the apoplast proteome after cold acclimation using quantitative proteomics. In total, 751 apoplast proteins were identified in jojoba, and the abundance of 200 proteins showed significant changes after cold acclimation. These proteins were primarily involved in defense, cell wall modification, carbohydrate metabolism, and redox balance. We also investigated the function and regulation of a cold acclimation-responsive class III chitinase ScCHIA. The results showed that the overexpression of ScCHIA enhanced the tolerance of Arabidopsis, yeast, and jojoba to low temperature and osmotic stress. Under cold stress, ScCHIA-overexpressing Arabidopsis accumulated more osmolytes, activated antioxidant enzymes, and reduced stomatal aperture size, which may contribute to enhanced tolerance to cold stress. ScCHIA was induced by methyl jasmonate (MeJA) application, and electrophoretic mobility shift assay (EMSA), yeast one-hybrid (Y1H), and luciferase activity assays demonstrated that an E-box cis-acting element on the ScCHIA gene promoter mediated regulation of ScCHIA by MeJA signaling, indicating that elevated levels of MeJA caused by cold acclimation may promote the expression of ScCHIA, thereby enhancing the cold tolerance of jojoba. Our research highlights the important role of the apoplast in plant response to low temperature stress and improves the understanding of the functions and regulatory mechanisms of plant chitinases in abiotic stress response.

Tetraena mongolica is a rare xerophytic shrub native to arid regions of western China. T. mongolica, commonly known as “oil firewood,” was once used as an important fuel source by local herders. To understand lipid accumulation and metabolism in the non-seed organs of T. mongolica, we analyzed lipid content in its roots, stems, and leaves. We identified 450 lipids, of which 78 were triglycerides (TGs), making it the most diverse subclass. Roots and stems accumulated significantly higher levels of total lipids and TGs than leaves, and both lipid fatty acids and total fatty acids showed similar accumulation patterns across different organs. A total of 38 fatty acids were detected in T. mongolica lipids, with C16:0, C18:1, and C18:2 being the dominant TG components across all organs. In the analysis of total fatty acids, hydroxy fatty acids and dicarboxylic acids emerged as the most abundant. By integrating the relative content of lipid fatty acids with transcriptome analysis, we identified 15 core genes involved in the lipid metabolism pathway and also identified genes with a high level of association with these pathways. The expression levels of these genes were determined to be closely correlated with the accumulation of relevant lipids, suggesting that they play important roles in the regulation of fatty acid and oil metabolism. This study presents a systematic analysis of lipid metabolism in the non-seed organs of T. mongolica, offering a new theoretical foundation for utilizing vegetative organs from extreme environments as alternative sources of plant oils and biofuels.

Jojoba (Simmondsia chinensis) is a commercially important woody oil crop whose productivity is limited by low-temperature sensitivity. Chitinases (CHIAs), members of the glycoside hydrolase family, play vital roles in plant defense and stress adaptation, yet their functions in jojoba remain unexplored. Here, 19 ScCHIA genes were identified in the jojoba genome, and phylogenetic analysis revealed a lineage-specific expansion of class III members. Among them, ScCHIA1 showed the strongest response, with transcript levels increasing by up to 2.07-fold under cold stress and 1.95-fold under osmotic stress. Functional assays demonstrated that ScCHIA1 overexpression enhanced stress tolerance in yeast and transgenic jojoba by promoting antioxidant activity and maintaining membrane stability. Furthermore, ScWRKY23 directly bound to the ScCHIA1 promoter and activated its transcription, and its overexpression in transgenic jojoba further improved stress resistance. Together, these findings reveal a novel ScWRKY23-ScCHIA1 regulatory module that mediates chitinase-associated stress responses and provide potential targets for the genetic improvement of stress tolerance in jojoba.

Plant pathogenesis-related (PR) proteins are a large and diverse family of proteins with antimicrobial activity, often induced by pathogen attack. Traditionally, PR proteins were thought to mainly participate in plant defense mechanisms against biotic stress. However, in recent years, increasing evidence has shown that these proteins also play important roles in the response to abiotic stress in plants. In the present review, we provide a summary of the latest findings on PR proteins and focus on their response to various abiotic stresses, the mechanism by which PR proteins are activated by external and internal signals, and their biological functions in plant responses to abiotic stresses. In addition, the existing challenges and future applications are also summarized, aiming to provide a reference for further research on PR proteins in the context of plant physiology.

Flavonols serves as bioactive pharmaceutical components in Astragalus membranaceus, and their accumulation is induced under drought stress. However, the underlying molecular mechanism remains unclear. In this study, transcriptome sequencing analysis revealed that drought stress activated flavonol biosynthesis in A. membranaceus, with AmMYB35 participating in regulation of flavonol biosynthesis. AmMYB35 is a SG7 group R2R3-MYB transcription factor. Constitutive expression of AmMYB35 increased flavonol content, upregulated the expression of the related enzyme genes, and improved drought tolerance in both Arabidopsis thaliana and A. membranaceus. Conversely, silencing AmMYB35 decreased flavonol accumulation and compromised drought tolerance in A. membranaceus. Further investigation revealed that AmMYB35 promoted the expression of flavonol synthase (FLS) by binding to its promoter. Collectively, these findings revealed that drought stress induced flavonol accumulation via the AmMYB35-AmFLS regulatory module, with elevated level of flavonol enhancing drought resistance in A. membranaceus by reducing ROS accumulation. This study offered novel insights into the influence of environmental factors on flavonol biosynthesis in A. membranaceus.

Flavonoids are rich active components in plants, widely used in pharmaceuticals and nutritional supplements. Flavonoids are the important active healthcare components in Astragalus membranaceus. Methyl jasmonate (MeJA) is an important growth regulator that promotes the accumulation of flavonoids in a variety of plants. However, the molecular regulatory mechanism of the flavonoid synthesis in A. membranaceus after MeJA treatment is unclear. We conducted metabolomic and transcriptomic analyses of A. membranaceus subjected to MeJA treatment were conducted. A group of 118 differentially accumulated metabolites (DAMs) identified were primarily associated with the biosynthesis pathways of flavones. Flavonoid biosynthesis was one of the most enriched pathways in differentially expressed genes (DEGs). Multiple-omics analysis revealed an association between gene expression levels and the metabolic flux within the flavonoid pathway. Weighted gene co-expression network analysis (WGCNA) identified five co-expression modules and nineteen hub genes involved in flavonoid biosynthesis. AmMYB30 interacted with the promoters of AmCHS, AmFLS, and AmF3H, thereby regulating the flavonoid signaling pathway in A. membranaceus. Elevated expression of AmMYB30 in hairy roots resulted in heightened levels of AmCHS, AmFLS, and AmF3H expression, along with an increase in total flavonoid content, indicated AmMYB30 induced flavonoid accumulation after MeJA treatment. In summary, this study identified key regulators for flavonoid biosynthesis and offered valuable insights for understanding the regulation of flavonoid biosynthesis in A. membranaceus.

Chitinases are enzymes that hydrolyze β-1,4-glycosidic bonds in chitin. Previous studies have shown that several chitinases accumulated significantly in A. mongolicus, suggesting that chitinases might participate in the adaptation to winter climate in Ammopiptanthus mongolicus. Here, we analyzed the evolution and expression patterns of the chitinase gene family in A. mongolicus and investigated the function and regulatory mechanisms of the AmChi7 gene in response to abiotic stress. The chitinase gene family in A. mongolicus comprises 27 members, many of which arose through formed by tandem and segmental duplication. Several chitinase genes, including AmChi7 gene, were significantly upregulated in winter. Overexpression of AmChi7 gene enhanced the tolerance of yeast to freeze-thaw cycle and osmotic stress, and enhanced the tolerance of transgenic Arabidopsis to low-temperature and drought stress. Furthermore, AmWRKY59, a MeJA-induced transcription factor, bound to the W box element in the AmChi7 gene promoter, activating its expression in winter. It is speculated that chitinase AmChi7 accumulation in winter enhances adaptation to temperate winter climates in A. mongolicus. This study expands our understanding of the biological functions of chitinases and provides insights into the molecular mechanisms underlying winter climate adaptation in A. mongolicus.

Jojoba (Simmondsia chinensis Schneider), the sole species in the Simmondsiaceae family, holds significant economic value. Understanding the mechanism of seed germination and post-germination is essential for jojoba breeding programs. For the first time, this study employed matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) to comprehensively detect and image endogenous lipids in jojoba seeds during germination and post-germination. Meanwhile, a new method of lithium salt-doped 2-mercaptobenzothiazole (2-MBT) matrix was adopted to detect the wax ester in jojoba seeds. The spatial lipidomic analysis of MALDI-MS revealed the dynamics of lipid alterations during three stages of jojoba seed (stage I, quiescent seed; stage II, germination; stage III, post-germination). MALDI-MSI results demonstrated heterogeneous distributions of differential lipids and main wax esters (WEs) in unique tissue regions of the jojoba seed at different stages. For instance, at stages II and III, three phosphatidylcholines (PCs) (PC(36:2), PC(36:4), and PC(35:3)) are mainly localized in the cotyledons, while the other two PCs (PC(34:1) and PC(36:1)) were distributed in both cotyledons and embryonic axis. In contrast, three phosphatidylethanolamines (PE(36:1), PE(42:4), and PE(42:9)) are primarily distributed near the embryonic axis and phosphatidylserine (PS(32:0)) was found exclusively in the embryonic axis area at three stages. In addition, three phosphatidic acids (PA(40:7), PA(46:2), and PA(42:1)) displayed highly accumulated in the middle of the cotyledons at stages II and III. Besides, WEs are continuously degraded mainly in the cotyledons during germination and post-germination processes. This study offers new insights into lipid alterations from a spatial lipidomics perspective and can contribute to our understanding of the role of these specific lipids during jojoba seed germination and post-germination. These findings will deepen our understanding of the metabolic processes involved in jojoba seed germination and seedling development and have underlying implications for crop improvement and seed quality control.

The Dehydration-Responsive Element Binding (DREB) subfamily of transcription factors plays crucial roles in plant abiotic stress response. Ammopiptanthus nanus (A. nanus) is an eremophyte exhibiting remarkable tolerance to environmental stress and DREB proteins may contribute to its tolerance to water deficit and low-temperature stress. In the present study, an A. nanus DREB A5 group transcription factor gene, AnDREB5.1, was isolated and characterized in terms of structure and function in abiotic stress tolerance. AnDREB5.1 protein is distributed in the nucleus, possesses transactivation capacity, and is capable of binding to DRE core cis-acting element. The transcription of AnDREB5.1 was induced under osmotic and cold stress. Tobacco seedlings overexpressing AnDREB5.1 displayed higher tolerance to cold stress, osmotic stress, and oxidative stress compared to wild-type tobacco (WT). Under osmotic and cold stress, overexpression of AnDREB5.1 increased antioxidant enzyme activity in tobacco leaves, inhibiting excessive elevation of ROS levels. Transcriptome sequencing analysis showed that overexpression of AnDREB5.1 raised the tolerance of transgenic tobacco seedlings to abiotic stress by regulating multiple genes, including antioxidant enzymes, transcription factors, and stress-tolerant related functional genes like NtCOR413 and NtLEA14. This study provides new evidence for understanding the potential roles of the DREB A5 subgroup members in plants.

Thaumatin-like proteins (TLPs) are conserved proteins involved in the defense and stress responses of plants. Previous studies showed that several TLPs were accumulated in leaf apoplast in Ammopiptanthus mongolicus in winter, indicating that TLPs might be related to the adaptation to winter climate in A. mongolicus. To investigate the roles of TLPs in winter adaptation, we first analyzed the expression pattern of TLP genes in A. mongolicus and then focused on the biological function and regulation pathway of AmTLP25 gene. Several TLP genes, including AmTLP25, were upregulated during winter and in response to both cold and osmotic stress. Overexpression of the AmTLP25 gene led to an increased tolerance of transgenic Arabidopsis to freezing and osmotic stress. Furthermore, the elevated AmWRKY14 transcription factor during winter activated AmTLP25 gene expression by specifically binding to its promoter. It is speculated that the AmWRKY14 - AmTLP25 module contributes to the adaptation to temperate winter climate in A. mongolicus. Our research advances the current understanding of the biological function and regulatory pathway of TLP genes and provides valuable information for understanding the molecular mechanism of temperate evergreen broad-leaved plants adapting to winter climate.

Plant cuticular wax forms a hydrophobic structure in the cuticle layer covering epidermis as the first barrier between plants and environments. Ammopiptanthus mongolicus, a leguminous desert shrub, exhibits high tolerances to multiple abiotic stress. The physiological, chemical, and transcriptomic analyses of epidermal permeability, cuticular wax metabolism and related gene expression profiles under osmotic stress in A. mongolicus leaves were performed. Physiological analyses revealed decreased leaf epidermal permeability under osmotic stress. Chemical analyses revealed saturated straight-chain alkanes as major components of leaf cuticular wax, and under osmotic stress, the contents of total wax and multiple alkane components significantly increased. Transcriptome analyses revealed the up-regulation of genes involved in biosynthesis of very-long-chain fatty acids and alkanes and wax transportation under osmotic stress. Weighted gene co-expression network analysis identified 17 modules and 6 hub genes related to wax accumulation, including 5 enzyme genes coding KCS, KCR, WAX2, FAR, and LACS, and an ABCG transporter gene. Our findings indicated that the leaf epidermal permeability of A. mongolicus decreased under osmotic stress to inhibit water loss via regulating the expression of wax-related enzyme and transporter genes, further promoting cuticular wax accumulation. This study provided new evidence for understanding the roles of cuticle lipids in abiotic stress tolerance of desert plants.

Apocynum venetum is a natural bast fiber crop and officinal plant belonging to the family Apocynaceae. Here, a chromosome -scale genome sequence was assembled using the single molecule sequencing and Hi -C technology. The genome of A. venetum is 228.80 Mb in length and has an N50 of 3.43 Mb. Up to 99.82 % of the contigs were attached to 11 pseudo -chromosomes. A total of 21,327 protein -coding genes were annotated in A. venetum genome. Phylogenetic analysis revealed the evolutionary relationships among the genera of Apocynum, Calotropis, and Catharanthus. Genes involved in bast fiber synthesis were identified in A. venetum by comparing gene expression between stem barks in different locations on the main stem. Combined metabolomic and transcriptomic analysis revealed the coordinated changes in flavonoids and related genes under methyl jasmonate treatment, as well as the dynamic change patterns of the genes encoding enzymes in the biosynthesis of flavonoids. Co -expression network analysis identified key genes that regulate flavonoid biosynthesis, including CHI (Av02G001490 and Av02G016020), MYB (Av01G029390), and bHLH (Av08G002930). The genome assembly of A. venetum provides a useful data set for understanding the evolution of Apocynum, bast fiber synthesis, and flavonoid biosynthesis in A. venetum.

Jojoba is an industrial oil crop planted in tropical arid areas, and its low-temperature sensitivity prevents its introduction into temperate areas. Studying the molecular mechanisms associated with cold acclimation in jojoba is advantageous for developing breeds with enhanced cold tolerance. In this study, metabolomic analysis revealed that various flavonols accumulate in jojoba during cold acclimation. Time-course transcriptomic analysis and weighted correlation network analysis (WGCNA) demonstrated that flavonol biosynthesis and jasmonates (JAs) signaling pathways played crucial roles in cold acclimation. Combining the biochemical and genetic analyses showed that ScMYB12 directly activated flavonol synthase gene (ScFLS). The interaction between ScMYB12 and transparent testa 8 (ScTT8) promoted the expression of ScFLS, but the negative regulator ScJAZ13 in the JA signaling pathway interacted with ScTT8 to attenuate the transcriptional activity of the ScTT8 and ScMYB12 complex, leading to the downregulation of ScFLS. Cold acclimation stimulated the production of JA in jojoba leaves, promoted the degradation of ScJAZ13, and activated the transcriptional activity of ScTT8 and ScMYB12 complexes, leading to the accumulation of flavonols. Our findings reveal the molecular mechanism of JA-mediated flavonol biosynthesis during cold acclimation in jojoba and highlight the JA pathway as a promising means for enhancing cold tolerance in breeding efforts.

The NAC family of transcription factors (TFs) is recognized as a significant group within the plant kingdom, contributing crucially to managing growth and development processes in plants, as well as to their response and adaptation to various environmental stressors. Ammopiptanthus mongolicus, a temperate evergreen shrub renowned for its remarkable resilience to low temperatures and drought stress, presents an ideal subject for investigating the potential involvement of NAC TFs in stress response mechanisms. Here, the structure, evolution, and expression profiles of NAC family TFs were analyzed systematically, and a cold and osmotic stress-induced member, AmNAC24, was selected and functionally characterized. A total of 86 NAC genes were identified in A. mongolicus, and these were divided into 15 groups. Up to 48 and 8 NAC genes were generated by segmental duplication and tandem duplication, respectively, indicating that segmental duplication is a predominant mechanism in the expansion of the NAC gene family in A. mongolicus. A considerable amount of NAC genes, including AmNAC24, exhibited upregulation in response to cold and osmotic stress. This observation is in line with the detection of numerous cis-acting elements linked to abiotic stress response in the promoters of A. mongolicus NAC genes. Subcellular localization revealed the nuclear residence of the AmNAC24 protein, coupled with demonstrable transcriptional activation activity. AmNAC24 overexpression enhanced the tolerance of cold and osmotic stresses in Arabidopsis thaliana, possibly by maintaining ROS homeostasis. The present study provided essential data for understanding the biological functions of NAC TFs in plants.

Background: Ammopiptanthus mongolicus is a rare temperate evergreen shrub with high tolerance to low temperature, and understanding the related gene expression regulatory network can help advance research on the mechanisms of plant tolerance to abiotic stress. Methods: Here, time-course transcriptome analysis was applied to investigate the gene expression network in A. mongolicus under low temperature stress. Results: A total of 12,606 differentially expressed genes (DEGs) were identified at four time-points during low temperature stress treatment, and multiple pathways, such as plant hormones, secondary metabolism, and cell membranes, were significantly enriched in the DEGs. Trend analysis found that the expression level of genes in cluster 19 continued to upregulate under low temperatures, and the genes in cluster 19 were significantly enriched in plant hormone signaling and secondary metabolic pathways. Based on the transcriptome data, the expression profiles of the genes in abscisic acid, salicylic acid, and flavonoid metabolic pathways were analyzed. It was found that biosynthesis of abscisic acid and flavonoids may play crucial roles in the response to low temperature stress. Furthermore, members of the phenylalanine ammonia-lyase (PAL) family in A. mongolicus were systematically identified and their structures and evolution were characterized. Analysis of cis-acting elements showed that the PAL genes in A. mongolicus were closely related to abiotic stress response. Expression pattern analysis showed that PAL genes responded to various environmental stresses, such as low temperature, supporting their involvement in the low temperature response in A. mongolicus. Conclusions: Our study provides important data for understanding the mechanisms of tolerance to low temperatures in A. mongolicus.

Ammopiptanthus mongolicus is an evergreen broad-leaved shrub growing in temperate regions. Plant defensins, a type of cysteine-rich small peptides, contribute to plant defense as antimicrobial peptides. In this study, we analyzed the evolution and expression patterns of the defensin gene family in A. mongolicus, and explored the function and regulatory mechanisms of the AmDEF2.7 gene in response to abiotic stress. Seven out of ten defensin genes had undergone segmental duplication and tandem duplication, especially, AmDEF2.6, AmDEF2.7, and AmDEF2.8, which were clustered on chromosome 9. The expression of multiple defensin genes was responsive to abiotic stress, with three defensin genes, including AmDEF2.7, showing significant induction during winter. Yeast expressing AmDEF2.7 gene exhibited increased resistance to freeze-thaw cycles and osmotic stress, while transgenic Arabidopsis overexpressing the AmDEF2.7 showed improved tolerance to both freezing and drought conditions. Furthermore, AmWRKY14 bound to the AmDEF2.7 gene promoter, and activated the expression of AmDEF2.7. These results highlighted the role of defensin AmDEF2.7 in the adaptation of A. mongolicus to temperate winter climate. This study expands our knowledge of plant defensin and provides support for clarifying the molecular mechanism of the adaptation of A. mongolicus to winter climate.