Global warming leads to premature dormancy release and untimely flowering in southern highbush blueberry during winter, resulting in chilling injury and yield losses. However, effective strategies to delay flowering by modulating dormancy progression without compromising fruit quality remain lacking. This study demonstrated through field trials that spraying 1 mg/mL ethephon (ETH) during the early endodormancy stage effectively delayed dormancy release and reduced the bud break rate of spring shoots by approximately 33.92% relative to the control, with no adverse effects on fruit quality. The treatment also reduces sucrose content in floral buds, a change potentially associated with dormancy maintenance. To explore the molecular basis of this process, we examined two ethylene-responsive transcription factors, VcERF112 and VcERF115, previously identified in our laboratory. Their expression was rapidly upregulated following ETH treatment. Heterologous expression of either gene in Arabidopsis delayed both seed germination and flowering, suggesting a conserved growth-suppressive function. Dual-luciferase reporter assays confirmed that VcERF112 and VcERF115 bind to the T2 region (−2310 to −1595 bp) of the VcBRC1 (VcBRANCHED1) promoter and enhance its expression. In contrast, sucrose treatment suppressed VcBRC1 expression. Collectively, these results propose that ethylene may sustain bud dormancy through a coordinated mechanism that operates independently of the classic abscisic acid (ABA)/gibberellins (GA) balance, a relationship not addressed in this study. This mechanism involves the induction of VcERF112/115 to activate VcBRC1, coupled with the reduction in sucrose levels to alleviate its repressive effect on VcBRC1. These findings provide new molecular insights into the ethylene-mediated regulatory network underlying bud dormancy in blueberry.

Highbush blueberry (Vaccinium corymbosum) is an important horticultural crop of significant nutritional, therapeutic, and economic value. However, the development of elite cultivars via genetic transformation has been severely restricted by labor-intensive tissue culture requirements and low transformation efficiency. Here, a simple, efficient, and tissue culture-independent Agrobacterium rhizogenes-mediated hairy root transformation system was established for the highbush blueberry cultivar "Bluerain." By eliminating the need for aseptic manipulation, new shoots generated from semilignified stem segments were subjected to vacuum-assisted A. rhizogenes strain K599 infiltration. Approximately 3 months post-infiltration, high biomass hairy roots was successfully induced, and the transformation efficiency reached 74.4% following stepwise optimization. This in planta protocol was further successfully applied to the cultivars "O'Neal," "Legacy," and "Emerald," yielding transformation efficiencies range from 26.7% to 40.0%. This optimized genetic transformation approach provides a valuable tool for the functional characterization of root-specific gene and for accelerating clonal propagation and genetic improvement in highbush blueberry.

Flavonoids in plant extracts exhibit significant biological activities but are limited by their low bioavailability. This research aimed to improve the functional properties by synthesizing polyvinylpyrrolidone (PVP) nano- particles using flavonoids derived from T. hemsleyanum. 208 flavonoids were identified using ultrahighperformance-mass/mass spectrometry and used for nanoparticle preparation. The nanoparticles exhibited 24 % and 55 % higher equilibrium dissolution rates in simulated intestinal and gastric fluids than free flavonoids. With spherical morphologies, 100 nm size and- 26.7 mV zeta potential, these nanoparticles showed remarkable improvements in biological activities, including 1.5-fold enhancement in antioxidant capacity, HeLa cell inhibition, and antimicrobial efficacy, and a 5-fold increase in anti-inflammatory effects compared to unencapsulated flavonoids. This research has established a methodological system that could effectively enhance the utilization rate of flavonoids in plant extracts, offering promising potential for application in functional foods.

Cervical cancer progresses through distinct precancerous stages, making early screening and intervention crucial for prevention. However, conventional screening modalities, such as cytology and HPV testing, face challenges related to sensitivity, specificity, and resource dependency. Circular RNAs (circRNAs), owing to their high stability and tissue-specific expression, have emerged as promising biomarkers, though their role in cervical carcinogenesis remains underexplored. In particular, the clinical utility of circRNAs for optimizing cervical cancer screening and early diagnosis has yet to be established. This study aimed to investigate the dynamic expression profiles of circRNAs across various stages of cervical cancer progression and identify potential biomarkers to enhance early detection. CircRNA sequencing was performed on cervical tissues spanning normal cervical epithelium (NCE), high-grade squamous intraepithelial lesions (HSIL), and cervical squamous cell carcinoma (CSCC). Functional assays, including cell viability, colony formation, and apoptosis, were performed to assess the oncogenic potential of circPOLD1 and its interaction with YBX1 in cervical cancer cells. BaseScope and immunohistochemistry (IHC) were applied to tissue microarrays for clincial validation and ROC curve analysis evaluated the diagnostic performance of circPOLD1 in serum as a liquid biopsy marker. CircRNA profiling revealed a progressive increase in circPOLD1 expression from NCE to HSIL and CSCC. Mechanistically, circPOLD1 functioned as an oncogene by binding to and phosphorylating YBX1, activating the AKT/mTOR/HIF-1α pathway to enhance glycolysis-driven tumorigenesis. BaseScope and IHC confirmed the stage-specific elevation of circPOLD1 and YBX1 in cervical lesions. The circPOLD1/YBX1 multi-marker panel demonstrated superior diagnostic performance, achieving an AUC of 0.951 for LSIL+ and 0.817 for HSIL+ detection. Furthermore, serum circPOLD1 levels exhibited a progressive increase across disease stages, underscoring its potential as a non-invasive biomarker. circPOLD1 and YBX1 synergistically drive cervical carcinogenesis and exhibit stage-specific expression patterns. Their combined detection significantly enhanced the accuracy for cervical cancer screening and dynamic monitoring. The successful application of BaseScope and IHC highlights the immediate translational potential of these biomarkers, paving the way for refined risk stratification, improved therapeutic targeting, and reduced cervical cancer burden through early intervention.

Chinese bayberry (Myrica rubra or Morella rubra) is a valuable fruit, yet the mechanism of its flesh segment development is not well understood. Using paraffin sectioning, we investigated the flower buds of the ‘Biqi’ and ‘Zaojia’ varieties, revealing that the flesh segment development in these Chinese bayberry varieties involved the formation of a primordium outside the ovary wall, the establishment of a simple columnar structure, and the formation of the primary flesh segment. Assessment of endogenous hormone levels indicated the significant reductions in jasmonic acid (JA) and indole-3-acetic acid (IAA) levels at the critical stages of flesh segment development. Correlation analysis highlighted the essential roles of IAA, JA, abscisic acid (ABA), and gibberellins in the flesh segment developmental process, underscoring the complex interactions driven primarily by the IAA, JA, and ABA networks. Gene modules positively correlated with flesh segment development were identified using transcriptome-based weighted gene co-expression network analysis (WGCNA). Differentially expressed genes (DEGs) were enriched in plant hormone signal transduction pathways, particularly for upregulated genes associated with auxin and JA signaling. Key genes predicted to be involved in flesh segment development included LAX2 and LAX3 (auxin transport), JAZ6 (JA signaling repression), and KAN1 and KAN4 (regulating multiple hormonal signaling pathways). Quantitative real-time polymerase chain reaction (qRT-PCR) validation confirmed that the expression trends for these genes were consistent across both varieties, particularly for CRC, SEP1, SEP3, IAA7, and JAZ6. Immunofluorescence localization studies revealed that auxin was primarily distributed in the central vascular bundle and outer cells of the flesh segment. This uneven auxin distribution might contribute to the unique morphology of flesh segments. Overall, this study provides insights into the hormonal regulation and genetic factors involved in the development of Chinese bayberry flesh segments.

The HECATH (HEC) gene, a significant member of the basic helix-loop-helix (bHLH) transcription factor family, exerts a crucial influence on pistil development through the regulation of auxin and cytokinin biosynthesis and transport. The gene plays a pivotal role in the development of the pistil. To investigate the functional and regulatory roles of VcHEC in blueberry bud dormancy release, this study selected flower buds from the southern highbush blueberry cultivar 'O'Neal' (complex hybrids of V. corymbosum and low-chill Vaccinium species) as experimental materials. The research combined qRT-PCR analysis, genetic transformation techniques, and yeast one-hybrid assays to conduct systematic investigations. The results revealed significant differences in the VcHEC gene during the critical period of dormancy release, with its expression pattern aligning consistently with PINFORMED3 (PIN3). Results further demonstrate that VcPIN3 is regulated by VcHEC, suggesting that the VcHEC gene may be involved in the release of blueberry bud dormancy by regulating the transport of auxin. The VcHEC gene enhanced the germination rate of Arabidopsis dormant seeds and facilitated plantlet development. Deletion of 5 ' fragments gradually decreased promoter activity, with proVcHEC showing much lower GUS staining intensity than p1VcHEC (from -1,775 bp to -1 bp) and fluorescence analysis demonstrated this. Screening 12 upstream regulatory genes using the VcHEC promoter (from -2,272 bp to -1,775 bp) as bait identified a negative correlation between squamosa promoter-binding-like (SPL) and VcHEC expression based on transcriptome data. It is speculated that VcSPL may serve as a key repressor of VcHEC gene expression, warranting further investigation into the specific relationship. This study not only enriches our understanding of the biological function of the VcHEC gene but also provides a novel perspective on the study of dormancy release mechanisms. In summary, the research on the role of the VcHEC gene in blueberry bud dormancy release provides a theoretical basis for enhancing blueberry cultivation, breeding, and cold resistance management. It also broadens our understanding of the dormancy mechanisms in perennial woody plant species.

Herein, a simple yet powerful label-free electrochemical sensing platform developed for dual detection of Pb2+ and malathion by ingeniously integrating Pb2+-dependent DNAzyme and malathion aptamer into a triplex DNA structure. The designed DNA complex not only served as dual recognition elements but also functioned as signal amplifiers through DNA walker activity triggered by target binding. When detecting Pb2+, the activated DNAzyme initiated cyclic cleavage of substrate DNA, thereby generating numerous G-quadruplex/hemin complexes that produced significantly enhanced electrochemical signal and achieved its signal-on detection. Following this, for malathion detection, the specific binding between the pesticide and its aptamer effectively disrupted the DNA walker assembly, consequently reducing signal output forming a signal-off detection mode. This innovative design successfully achieved highly sensitive and specific detection of both contaminants on a single platform through merely two operational steps while avoiding complex labeling procedures. Significantly, the sensor demonstrated consistently reliable performance in analyzing various challenging environmental samples including urban river water, agricultural water and soil samples. By combining nucleic acid recognition specificity with DNA walker signal amplification and straightforward operation, this advanced sensing platform presents remarkable potential for practical environmental monitoring of both Pb2+ and malathion.

Autumn shoot pruning is a significant production measure in Highbush blueberry (Vaccinium corymbosum L.) that promotes floral differentiation, but the mechanism is still unclear. To explore the role of carbohydrates in floral induction and differentiation promoted by short pruning, carbohydrate content, and the transport and metabolic-related genes were compared and analyzed. It is found that sucrose might be an important trigger for pruning to promote floral induction. Compared with only pruning the shoots, cutting half of the leaves significantly reduces the flower bud rate and delays the flower differentiation process, while further adding 1.5% sucrose spray alleviates this phenomenon partly. During floral induction, the soluble sugar content in buds increases significantly, while the sucrose content in leaf midvein and buds increases earlier. The starch and soluble sugar content in the stem decreases significantly. At the early flower bud morphological differentiation stage, a large number of starch granules accumulate in the buds, sucrose in leaves increases, glucose in buds decreases, and then fructose decreases. There is an upregulation of VcFT, VcSOC1, VcFUL, VcSUC, and VcINV expression in the leaf midvein, as well as an upregulation of VcFT, VcSOC1, VcTPS1, VcINV, and VcSWEET1 expression in buds during floral induction. At the same time, interaction between VcSOC1 and VcTPS1 proteins exists. The level of VcSOC1 and VcAMY can also be upregulated by VcTPS1. Sucrose plays an important role in blueberry floral induction through flowering factors and sugar-related genes, especially VcFT, VcSOC1, VcTPS1, VcSUC, and VcINV. The results of this study laid a strong foundation for further research on the regulation mechanism of flower bud differentiation in blueberry and other woody plants.

Bud dormancy is a crucial mechanism that allows perennial woody plants to withstand adverse environmental conditions, and gibberellin (GA) typically promotes the dormancy release of buds. DELLA protein is the core factor in the GA signaling pathway. This study identified 11 VcDELLA genes from the blueberry genome and performed a comprehensive bioinformatics analysis and prediction of the VcDELLA gene family. GA not only inhibits the dormancy entry of blueberry flower buds under low temperature and short-day conditions but also promotes the dormancy release of flower buds. During the dormancy release process, exogenous GA4+7 significantly increased the GA level and VcDELLA2/4/7/10/11 expression, while it significantly decreased VcDELLA3/6/8 expression. It is widely accepted that the GA-GID1-DELLA signaling module plays important roles in regulating the dormancy process of flower buds. In this study, Y2H and BiFC experiments showed that VcDELLA3/6 proteins could interact withVcGID1b/c, VcGID2, and VcSOC1 proteins. Regardless of the presence or absence of GA, VcGID1b/c and VcGID2 proteins could interact with VcDELLA3/6 proteins to form the VcGID1-VcDELLA-VcGID2 complex. In the leaves of VcDELLA3/6-silenced blueberry plants, VcDELLA3/6, VcGA3ox, VcGA20ox, and VcSOC1 expression was significantly decreased. Meanwhile, VcGID1b and VcGID1c expression was significantly increased. The leaf shedding rates of blueberry plants overexpressing VcDELLA3 and VcDELLA6 increased by 25 % and 33.33 %, respectively. In VcDELLA3/6-overexpressing plants, the GA level and VcDELLA3/6, VcGA2ox, VcGA3ox, VcGA20ox, VcGID2, and VcSOC1 expression were significantly increased, whereas VcGID1b/c expression levels were significantly decreased in VcDELLA6-overexpressing plants. Furthermore, exogenous GA effectively alleviated the deciduous phenotype of VcDELLA3/6 overexpressing blueberry plants and downregulated VcGA2ox, VcGA3ox, and VcGA20ox expression. It could be inferred that VcDELLA3/6 could regulate the transition of dormancy states in blueberry flower buds through interactions with GA biosynthesis (VcGA2ox, VcGA3ox, and VcGA20ox), GA signal transduction (VcGID1b/c and VcGID2), and the flowering related protein VcSOC1. Additionally, it is proposed that there might be a pathway with VcDELLA6 as the core to regulate the chilling-mediated dormancy release of blueberry flower buds. The formation of the GA4+7-VcGID1b/c-VcDELLA6-VcGID2 complex could activate VcSOC1 expression, thereby promoting the dormancy release of blueberry flower buds.

Blueberry flower buds cultivated in greenhouses develop during both autumn and spring, with floral induction being a critical process for flowering, influenced by environmental factors. This study aimed to clarify the regulatory mechanisms governing floral induction in greenhouse blueberries, focusing on the similarities and differences in flower bud differentiation between the spring and autumn seasons. Understanding these mechanisms is pivotal for enhancing blueberry production. In this study, we analysed the phenotypic characteristics associated with flower bud differentiation and observed that short photoperiods markedly affect the induction process. Transcriptomic analyses revealed distinct major metabolic pathways activated in autumn compared to spring. Seasonal variations in carbohydrate metabolism were also noted, with sucrose hydrolysis being prominent in autumn and sucrose synthesis prevailing in spring. The interplay between circadian rhythms and photosynthesis appeared to facilitate the allocation of sugars for bud development. Subsequent investigations underscored the sensitivity of VcCO3 to variations in photoperiod. Predominantly localised in the nucleus, VcCO3 facilitated floral induction in response to short photoperiods by activating the expression of downstream genes, including VcFT, VcLFY, VcAP3, and VcSOC1. Furthermore, VcCO3 exhibits a close association with the sugar metabolism gene VcSUS, promoting increased sucrose concentrations.

Mimetic recognition materials for rapid and selective analysis of foodborne contamination from complex foodstuff becomes a hot research topic in the field of food safety. In this work, borate-affinity-functionalized MOF/ molecularly imprinted polymer paper-based microfluidic chips (FSU-BA@MIP) were prepared by growing functionalized MOFs on paper surface with 3-carboxyphenylboronic acid (3-CPBA) as the ligands, followed by coating of a MIP layer of salbutamol (SAL) through a borate-affinity surface imprinting strategy. Using the obtained paper-based chip as a mimetic recognition and specific enrichment module, the fluorescence sensing platform achieved visual rapid quantitative analysis of SAL with a linear range of 0.1 x 10- 2-5.0 mg L- 1. The limits of detection and quantification were calculated to be 0.095 and 0.290 mu g kg- 1, respectively, and the sample loading via lateral flow aid could be completed within 30 min. The obtained MOFs/MIPs paper chips provide an effective way for the rapid analysis of harmful contaminants from different animal-derived foods.

The developmental plasticity of the maize inflorescence depends on meristems, which directly affect reproductive potential and yield. However, the molecular roles of upper floral meristem (UFM) and lower floral meristem (LFM) in inflorescence and kernel development have not been fully elucidated. In this study, we characterized the reversed kernel1 (rk1) novel mutant, which contains kernels with giant embryos but shows normal vegetative growth like the wild type (WT). Total RNA was extracted from the inflorescence at three stages for transcriptomic analysis. A total of 250.16-Gb clean reads were generated, and 26,248 unigenes were assembled and annotated. Gene ontology analyses of differentially expressed genes (DEGs) detected in the sexual organ formation stage revealed that cell differentiation, organ development, phytohormonal responses and carbohydrate metabolism were enriched. The DEGs associated with the regulation of phytohormone levels and signaling were mainly expressed, including auxin (IAA), jasmonic acid (JA), gibberellins (GA), and abscisic acid (ABA). The transcriptome, hormone evaluation and immunohistochemistry observation revealed that phytohormone homeostasis were affected in rk1. BSA-Seq and transcriptomic analysis also provide candidate genes to regulate UFM and LFM development. These results provide novel insights for understanding the regulatory mechanism of UFM and LFM development in maize and other plants.

Cross-pollination can improve the fruit set and quality of blueberry (Vaccinium spp.) for growers and consumers. However, the xenia effect in southern highbush blueberry remains unclear. Therefore, we selected eight cultivars of southern highbush blueberry (Vaccinium corymbosum L., interspecific hybrids) and applied pollination treatments (i.e., artificial self-pollination, artificial pollination with mixed pollen, or artificial pollination with individual cultivar pollen) to explore the xenia effects on the fruit set and quality of ‘O’Neal’ and ‘Emerald’. Pollen viability tests indicated that all of the cultivars are capable of fertilization. The highest fruit set came from ‘Bluerain’ pollen for ‘O’Neal’, while ‘Gulfcoast’ pollen increased fruit set the most in ‘Emerald’. Principal component analysis revealed that the cross combinations ‘Emerald’ × ‘Gulfcoast’ and ‘O’Neal’ × ‘Gulfcoast’ optimized the external and interior quality of the fruit. SSR was applied to determine which pollen source yielded the most seedlings. Results indicated that ‘Emerald’ × ‘Gulfcoast’ and ‘O’Neal’ × ‘Bluerain’ increased seedling production. Our results demonstrate that the xenia effects of ‘Gulfcoast’ pollen may increase ‘Emerald’ yields and promote fruit quality, while pollen from ‘Bluerain’ or ‘Jewel’ can improve ‘O’Neal’ fruit quality and seed number. Hence, these cross combinations may be utilized in blueberry production to increase fruit set, yield, and quality.

The coronavirus disease 2019 (COVID-19) pandemic is caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The spike protein (S) of SARS-CoV-2 is a major target for diagnosis and vaccine development because of its essential role in viral infection and host immunity. Currently, time-dependent responses of humoral immune system against various S protein epitopes are poorly understood. In this study, enzyme-linked immunosorbent assay (ELISA), peptide microarray, and antibody binding epitope mapping (AbMap) techniques were used to systematically analyze the dynamic changes of humoral immune responses against the S protein in a small cohort of moderate COVID-19 patients who were hospitalized for approximately two months after symptom onset. Recombinant truncated S proteins, target S peptides, and random peptides were used as antigens in the analyses. The assays demonstrated the dynamic IgM- and IgG recognition and reactivity against various S protein epitopes with patient-dependent patterns. Comprehensive analysis of epitope distribution along the spike gene sequence and spatial structure of the homotrimer S protein demonstrated that most IgM- and IgG-reactive peptides were clustered into similar genomic regions and were located at accessible domains. Seven S peptides were generally recognized by IgG antibodies derived from serum samples of all COVID-19 patients. The dynamic immune recognition signals from these seven S peptides were comparable to those of the entire S protein or truncated S1 protein. This suggested that the humoral immune system recognized few conserved S protein epitopes in most COVID-19 patients during the entire duration of humoral immune response after symptom onset. Furthermore, in this cohort, individual patients demonstrated stable immune recognition to certain S protein epitopes throughout their hospitalization period. Therefore, the dynamic characteristics of humoral immune responses to S protein have provided valuable information for accurate diagnosis and immunotherapy of COVID-19 patients.

Endodormancy is the stage that perennial plants must go through to prepare for the next seasonal cycle, and it is also an adaptation that allows plants to survive harsh winters. Blueberries (Vaccinium spp.) are known to have high nutritional and commercial value. To better understand the molecular mechanisms of bud dormancy release, the transcriptomes of flower buds from the southern highbush blueberry variety “O’Neal” were analyzed at seven time points of the endo- and ecodormancy release processes. Pairwise comparisons were conducted between adjacent time points; five kinds of phytohormone were identified via these processes. A total of 12,350 differentially expressed genes (DEGs) were obtained from six comparisons. Gene Ontology analysis indicated that these DEGs were significantly involved in metabolic processes and catalytic activity. KEGG pathway analysis showed that these DEGs were predominantly mapped to metabolic pathways and the biosynthesis of secondary metabolites in endodormancy release, but these DEGs were significantly enriched in RNA transport, plant hormone signal transduction, and circadian rhythm pathways in the process of ecodormancy release. The contents of abscisic acid (ABA), salicylic acid (SA), and 1-aminocyclopropane-1-carboxylate (ACC) decreased in endo- and ecodormancy release, and the jasmonic acid (JA) level first decreased in endodormancy release and then increased in ecodormancy release. Weighted correlation network analysis (WGCNA) of transcriptomic data associated with hormone contents generated 25 modules, 9 of which were significantly related to the change in hormone content. The results of this study have important reference value for elucidating the molecular mechanism of flower bud dormancy release.

This work mainly investigated the interferences of NaCl treatment to major metabolite accumulation, individual flavonoid biosynthesis and photosynthetic activity in Tetrastigma hemsleyanum. The results showed that the accumulation of phenolic acids, steroids and polysaccharides in T. hemsleyanum was significantly enhanced after the treatment of 50 or 100 mM NaCl for 12 days, and after 150 mM NaCl treatment for 3 days, the total content of flavonoids reached the highest value of 219.5 mu g.g(-1-1) and was higher by 3.8 times in comparison with the control (CK). In addition, the salt stress would generally enhance the reaction selectivity of Path II (RPSII) to produce flavonol derivatives (Rut, IsoQ, Km3rut, Ast, Qu, Km) and reduce the reaction selectivity of Path I (RPSI) to produce flavone derivatives (IsoO, Or, Ap), and after the treatment of 150 mM NaCl for 3 days, the RPSII and the RPSI reached the highest level of 0.785 and the lowest level of 0.215, respectively. Moreover, salt stress demonstrated significant and diverse effects on photosynthetic characteristics of T. hemsleyanum. Compared with CK, the values of Pn, Cond, Tr, Fv/Fm, Y(II) and qP increased first and then declined with increasing NaCl concentration and stress time, and the highest level of Pn, Fv/Fm and Y(II)occurred after 50 mM NaCl treatment. These findings clearly showed that 50 mM NaCl treatment results in favorable changes in photosynthesis and secondary metabolites accumulation in T. hemsleyanum.

Age has been found to be one of the main risk factors for the severity and outcome of COVID-19. However, differences in SARS-CoV-2 specific antibody responses among COVID-19 patients of different age groups remain largely unknown. In this study, we analyzed the IgG/IgM responses to 21 SARS-CoV-2 proteins and 197 peptides that fully cover the spike protein against 731 sera collected from 731 COVID-19 patients aged from 1 to 92 years. We show that there is no overall difference in SARS-CoV-2 antibody responses in COVID-19 patients in the 4 age groups. By antibody response landscape maps, we find that the IgG response profiles of SARS-CoV-2 proteins are positively correlated with age. The S protein linear epitope map shows that the immunogenicity of the S-protein peptides is related to peptide sequence, disease severity and age of the COVID-19 patients. Furthermore, the enrichment analysis indicates that low S1 IgG responses are enriched in patients aged <50 and high S1 IgG responses are enriched in mild COVID-19 patients aged >60. In addition, high responses of non-structural/accessory proteins are enriched in severe COVID-19 patients aged >70. These results suggest the distinct immune response of IgG/IgM to each SARS-CoV-2 protein in patients of different age, which may facilitate a deeper understanding of the immune responses in COVID-19 patients.

The basic leucine zipper (bZIP) is a transcription factor family that plays critical roles in abiotic and biotic stress responses as well as plant development and growth. A comprehensive genome-wide study in Liriodendron chinense was conducted to identify 45 bZIP transcription factors (LchibZIPs), which were divided into 13 subgroups according the phylogenetic analysis. Proteins in the same subgroup shared similar gene structures and conserved domains, and a total of 20 conserved motifs were revealed in LchibZIP proteins. Gene localization analysis revealed that LchibZIP genes were unequally distributed across 16 chromosomes, and that 4 pairs of tandem and 9 segmental gene duplications existed. Concluding that segmental duplication events may be strongly associated with the amplification of the L. chinense bZIP gene family. We also assessed the collinearity of LchibZIPs between the Arabidopsis and Oryza and showed that the LchibZIP is evolutionarily closer to O. sativa as compared to the A. thaliana. The cis-regulatory element analysis showed that LchibZIPs clustered in one subfamily are involved in several functions. In addition, we gathered novel research suggestions for further exploration of the new roles of LchibZIPs from protein-protein interactions and gene ontology annotations of the LchibZIP proteins. Using the RNA-seq data and qRT-PCR we analyzed the gene expression patterns of LchibZIP genes, and showed that LchibZIP genes regulate cold stress, especially LchibZIP4 and LchibZIP7; and LchibZIP2 and LchibZIP28 which were up-regulated and down-regulated by cold stress, respectively. Studies of genetic engineering and gene function in L. chinense can benefit greatly from the thorough investigation and characterization of the L. chinense bZIP gene family.