Iron-sulfur (Fe-S) clusters are essential cofactors for diverse biological processes, and their biosynthesis in chloroplasts is mediated by the evolutionarily conserved sulfur utilization factor (SUF) pathway. However, the molecular composition of the rice (Oryza sativa L., Os) SUF system and its roles in chloroplast biology and environmental stress adaptation remain unclear. Here, we report that mutations in the rice homolog of bacterial and plant SUFD, designated OsSUFD, lead to defective chloroplast development and premature leaf senescence. Biochemical analyses show that OsSUFD presumably assembles with OsSUFB and OsSUFC to form a chloroplastic scaffold complex. Notably, OsSUFC also interacts with a chloroplastic DNAJ chaperone, a potential iron donor for Fe-S cluster biosynthesis. Consistent with these observations, mutations in OsSUFD disrupt iron homeostasis and reduce the accumulation of plastidial Fe-S proteins. Furthermore, impaired OsSUFD function compromises rice tolerance to oxidative and heat stresses, accompanied by increased reactive oxygen species accumulation and decreased chlorophyll levels. In contrast, overexpression of OsSUFD in rice enhances tolerance to these abiotic stresses and alleviates the stress-induced physiological damage, further confirming its functional role in stress responses. Collectively, our findings define the conserved architectural features of the rice SUF system and reveal its essential roles in Fe-S cluster biogenesis, chloroplast function maintenance, and environmental stress tolerance.

The accumulation of root litter in soil can cause soil sickness, which negatively affect crop production. Here, we isolated bacterial and fungal strains capable of degrading tomato root litter and construct efficient cross-kingdom microbial consortium. Through in vitro screening, we obtained 16 strains, including five bacterial strains and eleven fungal strains that can degrade tomato root litter. The degradation abilities of the fungal strains were generally higher than that of the bacterial strains. Then, we successfully contrasted two cross-kingdom microbial consortiums Z2 (containing Aspergillus sp., Sarocladium strictum , Cellulosimicrobium funkei , and Penicillium sp.) and consortium Z3 (containing Aspergillus sp., Sarocladium strictum , Cellulosimicrobium funkei , and Microbacterium sp.) that could efficiently degrade tomato root litter. The microbial strains in each consortium were compatible to each other. These two microbial consortia showed higher degrading efficiency than any single strain in each consortium. The results indicate that fungi play a dominant role in the degradation of tomato root litters, and certain compatible strain consortia can enhance the degradation capacity.

Pre-harvest sprouting (PHS) significantly reduces the yield and quality of Chenopodium quinoa (quinoa). A key determinant of PHS resistance is the balance between seed dormancy and germination, a process primarily regulated by phytohormones. To elucidate the molecular mechanisms underlying hormone-mediated germination regulation, we performed transcriptome sequencing on Jingli 1 quinoa seeds 6 h after treatment with six phytohormones: abscisic acid (ABA), indole-3-acetic acid (IAA), jasmonic acid (JA), gibberellic acid (GA₃), brassinolide (BR), and 6-benzylaminopurine (6-BA). The results showed that ABA, IAA, and JA significantly inhibited germination, with a maximum inhibition rate of 36.73

Longan (Dimocarpus longan Lour.) is highly sensitive to low temperature, which severely restricts its cultivation and industrial development. MYB transcription factors serve as key regulators in plant responses to cold stress. In this study, an R2R3-MYB gene DlMYB108 was cloned from ‘Shixia’ longan. Sequence analysis showed that DlMYB108 contains two typical MYB repeats and shares high homology with cold-responsive MYB108 proteins from other plants. Expression pattern analysis revealed that DlMYB108 is highly expressed in young leaves, which are more sensitive to cold stress, and is significantly induced by low-temperature treatment. Subcellular localization and transcriptional activation assays confirmed that DlMYB108 is a nuclear-localized transcriptional activator. Yeast one-hybrid and dual-luciferase assays demonstrated that DlMYB108 specifically binds to the promoters of DlCBF2 and DlCBF3 and activates their transcription. Heterologous expression of DlMYB108 in Arabidopsis significantly enhanced cold tolerance, accompanied by reduced ion leakage, malondialdehyde (MDA) content and reactive oxygen species (ROS) accumulation, as well as upregulated expression of CBF and cold-responsive genes. Collectively, DlMYB108 positively regulates longan cold tolerance through activating DlCBF2 and DlCBF3 expression, providing a valuable candidate gene for cold-tolerant longan breeding.

RNA modification is crucial for the post-transcriptional regulatory mechanism that plays a pivotal role in determining RNA structure and function. Among these, N6-methyladenosine (m6A) represents the most abundant one in eukaryotic mRNA. In plants, m6A modification is catalyzed by a complex comprising multiple methyltransferase components. In this study, bioinformatic analyses were employed to characterize the genes of m6A methyltransferases (m6A writers), including their physicochemical properties, structures, cis-acting elements, chromosomal distributions, phylogenetic relationships, and predicted protein structures. Moreover, qRT-PCR and LC-MS/MS were utilized to investigate the expression patterns of m6A writer genes as well as the m6A abundance in total RNA from rice seedlings under low-temperature conditions. Additionally, m6A me-RIP sequencing was performed to explore changes in the m6A profile of mRNA in rice under cold stress. Collectively, our findings revealed the involvement in the regulation of mRNA m6A modification under cold stress in rice.

Waterfowl fatty liver has high nutritional value and offers benefits to human health. While previous research on fatty liver has focused on individual organs, this study explored fatty liver by examining the cecum, serum metabolites, and liver gene expression. This study integrated transcriptomic, metabolomic, and 16S rRNA microbiome analyses to analyze the molecular mechanism of waterfowl fatty liver formation. We identified seven core genes, five core metabolites, and three core microorganisms, which were significantly correlated. Overfeeding increased the abundance of Mucispirillum in the cecum, while Prevotella and Olsenella decreased. These microbial shifts, mediated by metabolites such as phthalic acid, influenced lipid metabolism, which induced changes in liver gene expression, including upregulation of ACBD4 and downregulation of HSP90B1 and HSPA5, thereby supporting fatty liver development. Additionally, ABC transporters, protein processing in the endoplasmic reticulum, and amino acid metabolism were important in fatty liver development. Our research findings provide new insights into the molecular mechanism of fatty liver with overfeeding in waterfowl from the perspective of the gut-liver axis.

With the intensification of climate change, spring low-temperature stress (SLTS) leads to floret degeneration and a decrease in grain number. This study investigated the physiological mechanisms underlying SLTS-induced floret degeneration using two wheat varieties with contrasting cold sensitivity. SLTS caused yellowing and shrinkage of floret primordia, increasing floret degeneration and fertile floret abortion, ultimately reducing grains per spike by 12.2%–26.1%. SLTS disrupted nutrient supply, impairing dry matter accumulation in young spikes. At 0–15 d after low-temperature stress (DALTS), SLTS caused a brief increase in the sugar content of young spikes (0–3 DALTS), followed by a rapid decrease (6–15 DALTS), while the total nitrogen content keeps decreasing. SLTS altered key enzyme activities, enhancing sucrose synthase and sucrose phosphate synthase but suppressing nitrate reductase and glutamine synthetase. Transcriptomic analysis revealed that SLTS perturbed starch and sucrose metabolism, carbon and nitrogen metabolism, and amino sugar pathways, altering soluble sugars, sucrose, fructose, and protein levels. SLTS disrupted carbon–nitrogen metabolic homeostasis, thereby reducing the number of fertile florets and ultimately leading to a decrease in grain number per spike. These findings enhance understanding of SLTS impacts on wheat floret development and provide insights for improving low-temperature tolerance and yield stability in wheat.

Endoplasmic reticulum (ER) stress and its associated unfolded protein response (UPR) have been demonstrated to play a crucial role in cancer’s progression, but their prognostic significance in breast cancer (BC) remains unclear. In this study, a reliable ER-related gene signature was developed for the purpose of predicting BC prognosis and investigating the associated immune landscape. By utilizing public datasets and analytical methods, we developed a 16 ER-related gene risk signature and verified its efficacy in predicting prognosis in independent patient groups. Patients in the high-risk group exhibited significantly poorer survival rates. Single-cell analysis revealed that the low-risk group exhibited stronger immune interactions. Conversely, the high-risk group exhibiting elevated immune checkpoints may signify an immunosuppressive microenvironment or heightened sensitivity to immune checkpoint inhibitor therapy. In vitro and vivo experiments confirmed that knocking down the expression of Marginal Zone B And B1 Cell Specific Protein (MZB1) significantly inhibited the proliferation, invasion, and tumorigenesis of breast cancer. The 16 ER-related gene signature is capable of effectively categorizing breast cancer patients into different risk levels, thereby providing a basis for personalized therapy. MZB1 has been identified as a significant regulatory factor, suggesting its potential as a target for the treatment of breast cancer.

Although heterosis plays a crucial role in enhancing crop yield and stress resistance, its underlying genetic mechanism remains not yet fully understood. Previous studies have shown that heterosis tends to increase with greater genetic distance in the absence of reproductive isolation barriers. However, whether variation in parental genome size alone can generate heterosis under near-isogenic backgrounds has not been thoroughly explored. Here, we used a rapeseed double haploid (DH) inducer line to generate progeny from the Pol CMS three-line hybrid Rongyou 18 (RY18). Although the progeny maintained the same ploidy level as the parents, their genome sizes showed notable variation (818.99–1024.88 Mb). To eliminate genetic distance effects, multiple DH progeny carrying restorer genes were crossed as paternal parents with the female parent 0068A of RY18, creating novel F1 hybrids. Using RY18 as the control, we observed a marked reduction in the genetic distance between the newly induced restorer line and the female parent (0068A). Correlation analysis further revealed a significant negative correlation (r = −0.310 *) between the paternal genome size and heterosis for thousand-seed weight (TSW). Furthermore, the genomic expansion in hybrid offspring relative to the male parent showed that significant correlations were observed between paternal genome size and heterosis over the standard for both TSW (r = 0.300, p < 0.05) and plot yield (r = 0.326, p < 0.05). Resequencing of high-and low-yielding F1 hybrids identified SNP sites, indicating that under an identical genetic background, heterosis for yield was more pronounced on chromosome A and chromosome C04. The doubled haploid (DH) induction line facilitates the generation of parental lines with distinct genome sizes, potentially providing a potential novel approach for studying heterosis research in Brassica napus.

The MEDIATOR (MED) complex is a molecular bridge that regulates transcription by connecting transcription factors (TFs) and specific gene promoters. The MED subunit 25 (MED25) can interact with specific TFs in the phytohormone signaling pathways to regulate the transcription of downstream genes and affect various physiological processes in plants. Although MED25 plays an important role in adaptation to abiotic stresses, it is still unclear whether MED25 is involved in freezing tolerance in wheat. In this study, we found that the expression of TaMED25 was significantly increased under extremely low temperatures (-25 degrees C) in Dongnongdongmai1 (Dn1) plants, a variety of winter wheat with excellent cold-stress resistance,and hypothesized that TaMED25 may be a novel regulator of freezing tolerance in this wheat variety. To further characterize the function of TaMED25 in freezing tolerance, we cloned TaMED25 from Dn1 plants and overexpressed it in Arabidopsis thaliana. The results showed that the overexpression of TaMED25 improved freezing tolerance in Arabidopsis plants, with decreased relative conductivity, enhanced ability to effectively scavenge reactive oxygen species (ROS), and increased expression of cold-responsive genes. Furthermore, we proposed that TaMED25 mediates jasmonate-induced cold resistance in plants, possibly by interacting with TaJAZ7 and TaMYC2 of the jasmonate signaling pathway and TaICE41 of the ICE-CBF-COR pathway. Our findings reveal the functions of MED25 in plants and provide a scientific basis for an in-depth analysis of the transcriptional regulation mechanism of freezing tolerance in winter wheat.

Mitogen-activated protein kinases (MAPKs) play important roles in plant stress response. As a major member of the MAPK family, MPK3 has been reported to participate in the regulation of chilling stress. However, the regulatory function of wheat (Triticum aestivum) mitogen-activated protein kinase TaMPK3 in freezing tolerance remains unknown. Dongnongdongmai No.1 (Dn1) is a winter wheat variety with strong freezing tolerance; therefore, it is important to explore the mechanisms underlying this tolerance. In this study, the expression of TaMPK3 in Dn1 was detected under low temperature and hormone treatment. Gene cloning, bioinformatics and subcellular localisation analyses of TaMPK3 in Dn1 were performed. Overexpressed TaMPK3 in Arabidopsis thaliana was obtained, and freezing tolerance phenotype observations, physiological indices and expression levels of ICE-C-repeat binding factor (CBF)-COR-related genes were determined. In addition, the interaction between TaMPK3 and TaICE41 proteins was detected. We found that TaMPK3 expression responds to low temperatures and hormones, and the TaMPK3 protein is localised in the cytoplasm and nucleus. Overexpression of TaMPK3 in Arabidopsis significantly improves freezing tolerance. TaMPK3 interacts with the TaICE41 protein. In conclusion, TaMPK3 is involved in regulating the ICE-CBF-COR cold resistance module through its interaction with TaICE41, thereby improving freezing tolerance in Dn1 wheat.

The fear response is a crucial adaptive mechanism for coping with environmental changes, and the individuals have different levels of fearfulness. The purpose of this study was to determine the status of the immune response and gut health in hens with different fear responses. A total of 80 healthy 75-wk-old native Lindian chickens were individually housed in conventional cages and categorized into high (TH) and low (TL) levels of fearfulness using the tonic immobility (TI) test. The immunological status and intestinal health of the laying hens were assessed, and the intestinal microbial community was sequenced using 16S rRNA testing. The results showed that the immune-related genes of interleukin (IL)-1β, IL-4, IL-6, and IgG were significantly upregulated in the spleen of TH hens compared with hens in the TL group (P < 0.01). The inflammatory immune-related genes Toll-like receptor (TLR)2, TLR4, nuclear factor (NF)-κB, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, IL-10, and IgG were significantly increased in the intestinal tract, whereas IL-4, IgA, and the intestinal barrier gene claudin-4 were significantly decreased in TH hens (P < 0.05). In addition, serum concentrations of IL-1β, IL-6, IL-10, interferon (IFN)-α and IgG were significantly higher in TH hens (P < 0.01). A high fear response also led to changes in gut microbial diversity, with a higher Simpson's index and lower β-diversity similarity than hens with a low-fear response (P < 0.05). The TH group showed an increase in 8 genera, including Bacillaceae and Coprococcus, whereas the genus Anaerorhabdus decreased (P < 0.05). The gut microbiota has also been associated with gut barrier genes, and inflammatory cytokines. Bartonella stimulates IL-1β and IgG secretion, whereas Lactobacillus inhibits IL-6 secretion, and Coprococcus and Subdoligranulum are associated with the maintenance of intestinal barrier function. The results of this study suggest that laying hens with high fear response levels have a more sensitive immune response and a more enriched gut microbiota, which may have positive effects on adapting to a complex environment.

Litters, the composition of sand and peat rich in microbiota, are essential to trigger the dustbathing behavior of chickens. To investigate the effects of a dustbathing environment (DE) on the intestinal health, gut microbiota, and immune responses of laying hens, a total of 72 healthy Hy-Line Brown laying hens at 69 wk of age (WOA) were housed individually in modified traditional cages and randomly divided into 2 groups: one group had free access to litters (CT), while the other one was restricted from litters (CC). The experiment lasted for 42 d. At the end of the experiment, the intestinal histomorphology and immune status of laying hens were determined, and the 16S rRNA sequencing method was used to assess the composition of the intestinal microbial community of birds. Intestinal histomorphology changed, including villus height and villus-to-crypt ratio significantly increased in the CT group (P < 0.01). DE reshaped the microbial community and increased the microbial richness with the higher indicators of Chao1 and observed species and the comparatively abundant beta diversity (P < 0.05). Ten genera, including Faecalibacterium and Coprococcus, declined in laying hens from the CT group (P < 0.05), while Alistipes increased in CT hens (P < 0.05) compared to those hens from the CC group. The expression levels of intestinal barrier-related genes of claudin-1, claudin-4, occludin, ZO-1, and ZO-2 and immune-related genes of IL-4, IL-6, IL-8, IFN-γ, IgA, TLR-2, and TLR-4 were significantly upregulated in the intestine of laying hens in CT group (P < 0.05). DE also increased the serum levels of IL-4, IL-6, IL-8, IFN-γ, and IgA (P < 0.01). The alteration of the gut microbiota by DE is closely related to host immune responses, including Lactobacillus positively correlated with IL-4 and IgA. Thus, a dustbathing environment can improve the welfare of laying hens by changing the intestinal histomorphology, immune response, and the gut microbial community.

Mitogen-activated protein kinases (MAPKs) play important roles in the stress response of plants. However, the function of MPK proteins in freeze-resistance in wheat remains unclear. Dongnongdongmai No.1 (Dn1) is a winter wheat variety with a strong freezing resistance at extremely low temperature. In this study, we demon-strated that TaMPK6 is induced by JA signaling and is involved in the modulation of Dn1 freeze resistance. Overexpression of TaMPK6 in Arabidopsis increased the survival rate of plant at-10 celcius. The scavenging ability of reactive oxygen species (ROS) and the expression of cold-responsive genes CBFs and CORs were significantly enhanced in TaMPK6-overexpressed Arabidopsis, suggesting a role of TaMPK6 in activating the ICE-CBF-COR module and antioxidant enzyme system to resist freezing stress. Furthermore, TaMPK6 is localized in the nu-cleus and TaMPK6 interacts with TaICE41, TaCBF14, and TaMYC2 proteins, the key components in JA signaling and the ICE-CBF-COR pathway. These results suggest that JA-induced TaMPK6 may regulate freezing-resistance in wheat by interacting with the TaICE41, TaCBF14, and TaMYC2 proteins, which in turn enhances the ICE-CBF-COR pathway. Our study revealed the molecular mechanism of TaMPK6 involvement in the cold resistance pathway in winter wheat under cold stress, which provides a basis for enriching the theory of wheat cold resistance.

Dongnongdongmai No.1 (Dn1) is one of the few winter wheat varieties that can successfully overwinter at temperatures as low as -25°C or even lower. To date, few researches were carried to identify the freeze tolerance genes in Dn1 and applied them to improve plant resistance to extreme low temperatures. The basic helix-loop-helix (bHLH) transcription factor MYC2 is a master regulator in JA signaling, which has been reported to involve in responses to mild cold stress (2°C and 7°C). We hypothesized that MYC2 might be part of the regulatory network responsible for the tolerance of Dn1 to extreme freezing temperatures. In this study, we showed that wheat MYC2 (TaMYC2) was induced under both extreme low temperature (-10°C and-25°C) and JA treatments. The ICE-CBF-COR transcriptional cascade, an evolutionary conserved cold resistance pathway downstream of MYC2, was also activated in extreme low temperatures. We further showed that overexpression of any of the MYC2 genes from Dn1 TaMYC2A, B, D in Arabidopsis led to enhanced freeze tolerance. The TaMYC2 overexpression lines had less electrolyte leakage and lower malondialdehyde (MDA) content, and an increase in proline content, an increases antioxidant defences, and the enhanced expression of ICE-CBF-COR module under the freezing temperature. We further verified that TaMYC2 might function through physical interaction with TaICE41 and TaJAZ7, and that TaJAZ7 physically interacts with TaICE41. These results elucidate the molecular mechanism by which TaMYC2 regulates cold tolerance and lay the foundation for future studies to improve cold tolerance in plants.

Footpad dermatitis (FPD) has a negative impact on the welfare and health production of poultry. The present study aimed to determine the effect of FPD on the expression of host defense peptides (HPDs). A total of fifty-four 55-week-old Hy-line brown hens housed individually in furnished cages were selected for investigating the mRNA expression of HPDs in foot, spleen, liver, and small intestine tissues. FPD score 0 group with normal foot, FPD score 1 with a small lesion of the foot pad epithelium and FPD score 2 with a larger lesion were divided by visual examination. The results showed that compared with group 0, hens from FPD group 1 and 2 had increased mRNA expression of HPDs of AvBD1, AvBD3, AvBD4, AvBD5, AvBD6, AvBD7, AvBD8, AvBD9, AvBD10, AvBD11, AvBD12, AvBD13, AvBD14 and LEAP2in foot and spleen tissues (P < 0.05). The relative expression of HPDs genes AvBD2, AvBD3, AvBD4, AvBD5, AvBD7, AvBD8, AvBD11, AvBD12, AvBD13, AvBD14, and LEAP2 in liver tissue increased, while the relative mRNA expression of AvBD6, AvBD9, and AvBD10 decreased (P < 0.05), and the expression levels of AvBD1, AvBD3, AvBD5, AvBD6, AvBD7, AvBD9, AvBD10, AvBD12, and LEAP2 in small intestinal tissue increased (P < 0.05), while the expression of AvBD2, AvBD8, and AvBD14 decreased (P < 0.05). Therefore, FPD can affect the expression of AVBDs in laying hens that the foot and spleen tissue had stronger responsiveness to the FPD. It is speculated that the increase in the expression of AVBDs on account of the possible infection with pathogenic bacteria. Then the increased HPDs participated in the host's defense responses and strengthened the immune regulation after FPD occurred.

Longan (Dimocarpus longan) is a typical southern subtropical fruit tree species that is sensitive to cold stress. C-repeat binding factors (CBFs), as transcription factors, are crucial components involved in the molecular regulation of the plant response to cold stress. However, the role of CBF homologs in the cold response regulation of longan remains largely unknown. Here, three novel CBF genes, DlCBF1, DlCBF2, and DlCBF3, were cloned from longan. DlCBF1 and DlCBF2 contain an AP2 domain and PKKPAGR and DSAWR CBF signature motifs, while DlCBF3 has mutations within these conserved signature motifs. DlCBF1/2/3 were mainly localized in the nucleus and specifically bound to CRT/DRE cis-elements, resulting in strong transcriptional activation. DlCBF1/2 exhibited tissue expression specificity, and their expression was induced by low temperature, while DlCBF3 had no tissue specificity and barely responded to low temperature. DlCBF1, DlCBF2, and DlCBF3 overexpression in Arabidopsis-enhanced cold tolerance by increasing proline accumulation and reducing reactive oxygen species (ROS) content, accompanied by upregulated expression of cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47, and AtKIN1) in the CBF cold stress response signaling pathway. In conclusion, the biological functions of DlCBF1/2/3 were somewhat conserved, but slow expression of DlCBF1/2 and low expression of DlCBF3 may partly cause the cold sensitivity of longan. Collectively, these results indicated that differences exist in the expression and function of CBF orthologs in the cold-sensitive plant species longan, and these findings may help to improve the understanding of the cold response regulation mechanism and provide important theoretical support for cold-tolerant breeding of longan.

BACKGROUND Soybean pod borer (Leguminivora glycinivorella) is an important soybean pest in north-eastern Asia, whose mature larvae overwinter in a diapause state. Disruption of winter diapause may be a valuable tool in pest management. However, the molecular mechanisms regulating diapause in this species have not yet been elucidated. RESULTS We compared the transcriptomes and proteomes between diapause and mature larvae and between mature and newly developed pupae to identify the genes and proteins associated with diapause. Thirty-seven differentially expressed genes and their proteins changed synchronously between diapause and mature larvae and 82 changed synchronously between diapause larvae and newly developed pupae. Among these, genes involved in fatty acid biosynthesis and the longevity regulating pathway were up-regulated in diapause larvae and down-regulated in newly developed pupae, suggesting that they may regulate diapause. One fatty acid synthase (FAS) gene and two small heat shock genes (HSP19.8andHSP18.9) were chosen for further functional analysis. After RNA interference (RNAi)-mediated knockdown ofFAS, the survival of mature larvae was significantly lower than that of control larvae, but the mean developmental time from first-instar larva to adult remained unchanged. RNAi-mediated knockdown ofHSP19.8andHSP18.9severely shortened the mean developmental time, causing approximately 50% larvae to develop directly into pupae. CONCLUSION FASand the small heat shock gene play roles in diapause regulation and larvae survival. This study provides important information that may assist in understanding the molecular regulatory mechanisms of overwintering diapause of this important agricultural insect pest.

ICE1 (inducer of CBF expression 1) encodes a typical MYC-like basic helix-loop- helix (bHLH) transcription factor that acts as a pivotal component in the cold signalling pathway. In this study, DlICE1, a novel ICE1-like gene, was isolated from the southern subtropical fruit tree longan (Dimocarpus longan Lour.). DlICE1 encodes a nuclear protein with a highly conserved bHLH domain. DlICE1 expression was slightly upregulated under cold stress. Overexpression of DlICE1 in Arabidopsis conferred enhanced cold tolerance via increased proline content, decreased ion leakage, and reduced malondialdehyde (MDA) and reactive oxygen species (ROS) accumulation. Expression of the ICE1-CBF cold signalling pathway genes, including AtCBF1/2/3 and cold-responsive genes (AtRD29A, AtCOR15A, AtCOR47 and AtKIN1), was also significantly higher in DlICE1-overexpressing lines than in wild-type (WT) plants under cold stress. In conclusion, these findings indicate that DlICE1 is a member of the bHLH gene family and positively regulates cold tolerance in D. longan.