Flavonoids play a critical role in plant resistance to various biotic and abiotic environmental stresses. Guizhou province is located in the subtropical monsoon climate zone with abundant but uneven precipitation, where spring drought frequently occurs and severely affects rice (Oryza sativa) sowing and seedling growth. In this study, the flavanone 3-hydroxylase (OsF3H) gene (GenBank No. XM_015776658) was cloned from rice leaves. Its CDS was 1 041 bp in length, encoding 346 amino acids, and the predicted protein molecular weight was 38.74 kD. Phylogenetic analysis revealed that OsF3H clusters with flavanone 3-hydroxylases from other species, indicating it was a member of the flavanone 3-hydroxylase family and shared the closest relationship with wheat (Triticum aestivum) F3H. Cis-acting element prediction indicated that the promoter region of this gene contained not only functional elements responsive to phytohormone signals, light regulation, as well as growth and development, but also cis-acting elements involved in drought and salt stress responses. Further expression analysis under drought treatment demonstrated that OsF3H could be induced by drought, suggesting its potential role in drought stress response of rice. To verify its function in drought response, OsF3H was heterologously expressed in Arabidopsis thaliana, and the transgenic plants were subjected to drought stress. The results showed that the transgenic plants exhibited significantly increased root length, greening rate, fresh weight, and dry weight relative to the wild-type plants, indicating that overexpression of OsF3H significantly enhanced drought tolerance of A. thaliana. These findings provide important data for further elucidating the physiological function of OsF3H in rice and its mechanism in response to drought stress, which will facilitate subsequent research.

Low-temperature saccharification refers to the process in which starch in the tubers of potato (Solanum tuberosum) is rapidly converted into reducing sugars when the potato is stored at low temperatures. It is a physiological response of potato to low-temperature stress, but severely impairs the quality of fried potato products. In order to reduce the accumulation of reducing sugars in potato tubers caused by low-temperature storage and improve the quality offried-potato products, this study used CRISPR/Cas9 technology to knock down the expression of the vacuolar invertase gene (StVInv), and created StVInv gene mutants. The results of qPCR and physiological and biochemical indicators testing indicated that after 21 d of storage at 4 ℃, the expression level of the StVInv gene in the mutant tubers was significantly lower than that in the wild type (P<0.0001). The vacuolar invertase activity was reduced by 33.68% to 60.4% compared with the wild type, and the reducing sugar content was decreased by 55.30% to 56.75% compared with the wild type. The color of the fried-potato chips was significantly improved compared with the wild type. This study provides technical support for the use of biotechnology to improve the deep-fried processing traits of potatoes.

Cembratriene-ol is a biodegradable green pesticide component that has significant application value in sustainable agriculture. However, the chemical synthesis of this substance is inefficient, and the extraction of the substance from plants is challenging. The precise mechanisms of biosynthetic processes and the identification of key intermediate metabolites remain to be elucidated. In this study, a multi-gene integration expression strategy was employed to heterologously express the geranylgeranyl diphosphate synthase (crtE) from Pantoea ananatis and cembratriene-ol synthase (cbts) from Nicotiana tabacum in Escherichia coli, while integrating the isopentenol utilization pathway (IUP) by introducing the choline kinase gene (chk) from Saccharomyces cerevisiae, the isopentenyl phosphate kinase gene (ipk), and E. coli's isopentenyl diphosphate isomerase gene (idi) to enhance the supply of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). SDS-PAGE analysis confirmed that the successful expression of all 5 target proteins in E. coli, with observed molecular weights aligning precisely with theoretical predictions. GC-MS analysis demonstrated the successful establishment of a heterologous E. coli synthesis pathway from isopentenol to cembratriene-ol, yielding a target product of 19.7 mg/L. Furthermore, the potential intermediate for cembratriene-ol synthesis from geranylgeranyl pyrophosphate (GGPP) was identified as cembrene, marking a significant discovery as it was the first time this intermediate was identified. This study provides fundamental evidence for elucidating the biosynthetic mechanism of cembratriene-ol, optimising its microbial synthesis pathway to promote the industrialisation of green pesticides, and also offers basic data for heterologous biosynthesis research on agricultural specialty secondary metabolites.

CLAVATA3/embryo surrounding region-related (CLE) genes are a class of gene widely distributed in plants that encode signaling peptides responses. In this study, CLE gene family members were identified from the whole genome of grape (Vitis vinifera) using bioinformatic approaches. Their physicochemical properties, phylogenetic relationships, gene structures and conserved motifs, chromosomal localization, and promoter cis-acting elements were systematically analyzed. At the same time, the expression levels of gibberellin (GA3) induced gibberellin in different organs were compared by combining transcriptome and qRT PCR. The results showed that a total of 15 CLE gene family members were present in the grape genome; they were unevenly distributed on 8 chromosomes, encoded proteins of 78~137 amino acids, and were classified into 4 subgroups. Subcellular localization predictions indicated that most VvCLE proteins were localized in the chloroplasts and nucleus, and all members contained the conserved CLE motif. Analysis of the promoter regions of VvCLE genes revealed multiple cis-acting elements associated with growth and development, hormone responses, light responsiveness, and stress adaptation. VvCLE genes were expressed in different organs of grape and exhibited obvious tissue specificity among them, VvCLE6 was highly expressed in the fruit flesh and was significantly induced by GA3, which was consistent with transcriptome data and suggested that it may be involved in fruit development. This study provides valuable resources and a theoretical basis for functional characterization and regulatory mechanism studies of the CLE gene family in grape.

Cucurbitacin B-type compounds in melon (Cucumis melo) possess important pharmacological effects such as anticancer and anti-inflammatory activities. Due to the high costs associated with their chemical synthesis and extraction, the mining and functional characterization of key genes involved in their biosynthetic pathways have attracted significant attention. To date, the key gene responsible for catalyzing the conversion of cucurbitacin B to cucurbitacin B-O-β-D-glucoside in melon remains unidentified. In this study, using the amino acid sequence of glycosyltransferases UDP-glycosyltransferase 74F2 (UGT74F2), which catalyzed the glycosylation of cucurbitacin E in watermelon (Citrullus lanatus), as a probe, the glycosyltransferases CmUGT6 gene in the melon genome with relatively high similarity was identified. Using the oriental melon variety 'Yumeiren' as experimental material, a 1 401 bp fragment of the CmUGT6 gene was cloned, encoding 467 amino acids. Bioinformatics analysis revealed that CmUGT6 contained a specific domain of the glycosyltransferase family, carried a negative charge, and possessed 46 phosphorylation sites, 3 N-glycosylation sites, and 4 O-glycosylation sites, with no transmembrane domain. The promoter region contained several cis-acting elements such as hormone responses, light responses, anaerobic induction, and metabolic regulation. Meanwhile, the CmUGT6 gene expressed in both the roots and fruits of the thin-skinned melon, with the highest expression level observed in the roots. Additionally, prokaryotic expression and purification of the CmUGT6 gene yielded a fusion protein band of approximately 100 kD. Molecular docking simulations between CmUGT6 and cucurbitacin B showed a binding energy of -8.9 kcal/mol. This study provides a reference for the subsequent in-depth exploration of the function of the CmUGT6 gene and the mechanism of glycosylation modification of cucurbitacin B in melon.

Phalaenopsis spp. has extremely high economic and ornamental value. The breeding of its new varieties generally adopts interspecific and intraspecific hybridization, resulting in complex genetic background and ploidy. In this study, expressed sequence tag-simple sequence repeat (EST-SSR) molecular marker technology was used to conduct genetic diversity analysis, hybrid identification, cluster analysis and ploidy prediction on 31 Phalaenopsis germplasm resources as well as the reciprocal F? hybrids of 'Treasure Map' and '075'. The aim was to provide a basis for the utilization and innovation of Phalaenopsis germplasm resources, molecular marker-assisted breeding, ploidy identification and authenticity identification of hybrid progeny. Based on 19 reported SSR primer pairs combined with the M13 universal primer, capillary fluorescence electrophoresis was performed. Ten pairs of EST-SSR primers were screened for genetic diversity, genetic relationship analysis and ploidy prediction of 31 germplasm resources of Phalaenopsis. Seven pairs of EST-SSR primers with paternal characteristic bands were screened for authenticity, cluster analysis and ploidy prediction of 50 F₁ hybrid progeny lines, and 50 F₁ hybrid progeny lines were identified by flow cytometry combined with chromosome counting. The results showed that a total of 233 alleles were detected in 10 pairs of primers, the average number of alleles detected at each locus was 23.3, the total number of effective alleles was 137.65, the average number of effective alleles at each locus was 13.76, and the polymorphism information content (PIC) value of 10 pairs of primers varied in the range of 0.32~0.91, only primer Pap-3754 had moderate polymorphism, and the other 9 primers had high polymorphism. The cluster analysis of 31 Phalaenopsis germplasm resources showed that the 31 Phalaenopsis germplasm resources were divided into 3 categories at the genetic similarity coefficient of 0.23, among which the classⅠand class Ⅱwere native to Phalaenopsis orchid, and the 25 materials of class Ⅲ were Phalaenopsis horticultural species. The authenticity of the hybrid F₁ progeny showed that all 50 hybrid F₁ progeny lines were identified as true hybrids. Cluster analysis of 50 crosses showed that the 49 of them tested were closely related to the maternal parent, and only one line, L25, was closely related to the paternal parent. Flow cytometry and chromosome counting results showed that the hybrid offspring of Phalaenopsis were tetraploid, and compared with the molecular marker results, the accuracy of EST-SSR molecular marker was as high as 98%. The results indicated that the EST-SSR molecular marker could be used for the analysis of genetic diversity, ploidy prediction and early identification of hybrid offspring of Phalaenopsis orchid. This study is of positive significance for improving the breeding efficiency of new varieties and exploring the genetic background of Phalaenopsis.

The new variety Lycoris albiflora 'Astro Girl' with a unique color and a temporal variation pattern, has extremely high ornamental value and is an ideal material for studying the mechanism of plant color gradient. To explore flower color formation mechanism in L. albiflora, R2R3-MYB transcription factor LaMYB56 gene associated with anthocyanin accumulation was cloned by RT-PCR. The regulation of anthocyanin accumulation was validated by molecular biology techniques. The results showed that the cDNA sequence of LaMYB56 with length of 1 031 bp and 723 bp ORF was obtained. LaMYB56 protein contains 2 conserved SANT domains at N-terminus. Phylogenetic analysis indicated that LaMYB56 was grouped with S20 subfamily in Arabidopsis thaliana. The expression level of LaMYB56 peaked on the first flowering day and gradually declined over time. The expression of LrF3'H, LrANS, LrGST genes were significantly decreased after silencing of LaMYB56 in the petals of L. radiata (P<0.05). Overexpression of LaMYB56 in Arabidopsis significantly increased the expression levels of AtUFGT and AtF3'H (P<0.05), leading to elevated anthocyanin content. The dual luciferase assay report and yeast one hybrid result indicated that LaMYB56 could bind to the promoter region of LaGST7 to regulate its expression and affect anthocyanin accumulation. The results demonstrated that LaMYB56 transcription factor was involved in the anthocyanins accumulation in L. albiflora. This study provides a theoretical found for the genetic improvement and molecular breeding of flower color in Lycoris genus and ornamental plants.

Plant root exudates are the key chemical signals and regulatory factors for plant response to environmental stress, and their composition changes directly affect the bioavailability of heavy metals in the rhizosphere and plant adaptability. In this study, using liquid chromatography-mass spectrometry (LC-MS) untargeted metabolomics, the metabolite changes in root exudates of Cynodon dactylon were systematically analyzed under soil Pb stress at concentrations of 500 and 1 000 mg/kg, and the tolerance mechanism of C. dactylon to soil Pb was revealed. The results showed that C. dactylon in the 500 mg/kg Pb treatment group exhibited higher root length, plant height, and leaf biomass compared with the 1 000 mg/kg Pb treatment group. Additionally, soil Pb stress reduced the contents of nitrogen, phosphorus and chlorophyll in leaves and roots of C. dactylon, indicating that soil Pb stress inhibited the absorption of nitrogen and phosphorus in soil and photosynthesis of C. dactylon. Different concentrations of soil Pb stress changed the metabolite components of C. dactylon root exudates, and a total of 1 028 differential metabolites were identified (VIP>1, P<0.05 ). The relative contents of DL-malic acid and γ-aminobutyric acid (GABA) in root exudates increased at 500 mg/kg soil Pb treatment group, but decreased at 1 000 mg/kg soil Pb treatment group. The metabolic response of C. dactylon to soil Pb stress revealed 2 key metabolic pathways associated with GABA: The GABAergic synapse pathway and butanoate metabolism pathway. These significantly enriched pathways under metal stress contributed to alleviating Pb-induced oxidative damage and ameliorating the imbalance in energy metabolism, and DL-malic acid had the ability to chelate heavy metals to reduce the bioavailability of Pb. Therefore, the organic acids (DL-malic acid, fumaric acid ) and GABA in the root exudates of C. dactylon jointly mediated the formation of a synergistic detoxification network of 'chelation-transport-antioxidation' to alleviate the stress of soil heavy metal Pb on plants. This study provides a theoretical basis for the regulation of plant rhizosphere metabolism on soil heavy metal Pb remediation technology.

Taxodium hybrid 'Zhongshanshan' relies on softwood cutting for asexual propagation. Elucidating the molecular mechanism of its adventitious root (AR) development and identifying key genes are of great significance for establishing an efficient genetic transformation system and improving propagation efficiency. To investigate the function of ethylene-insensitive 3 (EIN3), a core transcription factor in the ethylene (ET) signal transduction pathway of T. hybrid 'Zhongshanshan', during AR development, the ThEIN3 gene (GenBank No. PX209540) was cloned in this study. Its function was identified by overexpressing ThEIN3 in tobacco (Nicotiana benthamiana) through an Agrobacterium tumefacien-mediated expression system. The results showed that the full-length ThEIN3 gene was 2 073 bp, encoding 690 amino acids. It contained a typical conserved domain of the EIN3 superfamily and was localized in the nucleus. With the development of AR in T. hybrid 'Zhongshanshan', the expression level of ThEIN3 was gradually down-regulated, and its expression in mature soft shoots was significantly higher than that in rejuvenated soft shoots (P<0.05). Compared with wild-type tobacco (WT), the AR development of ThEIN3-overexpressing plants was inhibited. Hormone content measurements at the base of adventitious roots revealed a significant increase in the content of gibberellin A3 (GA3)(P<0.05), while the contents of indole-3-acetic acid (IAA) and trans-zeatin riboside (TZR) decreased. Further research on the molecular mechanism indicated that ThEIN3 might inhibit the occurrence of AR by coordinating key genes in the related hormone metabolism pathways. This study provides a foundation for further research on the regulatory mechanisms of AR development in T. hybrid 'Zhongshanshan' and offers genetic resources and theoretical guidance for its molecular breeding.

Vitrfication is commonly used for oocyte cryopreservation, however, cryoinjury and cryoprotectant toxicity results in damage to the structure and poor developmental capacity of oocytes after freezing. Liquid helium (LHe, -269 ℃) as cryogen, can improve cooling rate of vitrification. And high cooling rate is the key factor ensuring the success of vitrification of oocytes. In this study, laser tweezers Raman spectroscopy (LTRS) was used to non-invasively investigate the effects of varying vitrification temperatures (VTs) and cryoprotective agent concentrations (CPAs)(17.5% ethylene glycol (EG)+17.5% dimethyl sulfoxide (DMSO), 20% EG+20% DMSO) on the metabolic fingerprint profiles of bovine (Bos taurus) MⅡ stage oocytes after vitrification-thawing, and through in vitro developmental competence tests, the feasibility of using Raman spectroscopy to detect the quality of oocytes was explored. Bovine MⅡ oocytes were collected and randomized into 5 groups: Fresh oocytes (Fresh), oocytes vitrified in liquid helium including LHe-EDS35 and LHe-EDS40, oocytes vitrified in liquid nitrogen (LN, -196 ℃) including LN-EDS35 and LN-EDS40. The fresh oocytes and 4 experimental groups oocytes after vitrification-thawing were cultured in vitro for 4 h, then the culture media for single oocyte in different experimental groups were collected and separately analyzed using LTRS. Subsequently, they were evaluated embryo developmental competence after in vitro fertilization. These results showed that the significant changes were observed in the Raman spectra of culture media for oocyte after vitrification-thawing (P<0.05). The intensities of the characteristic peaks at 862 cm^-1 (tyrosine), 1 631 cm^-1 (Amide I: β-sheet), and 1 451 cm^-1 (lipids) for culture media of oocytes in LHe-EDS35, LHe-EDS40 were significantly lower than those in LN-EDS35, LN-EDS40 groups (P<0.05). Furthermore, vitrification induced a transformation of the protein secondary structure from the α-helices to the β-sheet form in oocytes of LN-EDS35 group. The results of in vitro development of oocytes from 5 experimental groups showed that cleavage rate, blastocyst rate of oocytes in LHe-EDS35 were significantly higher than those in LN-EDS35, LN-EDS40, LHe-EDS40 groups (P<0.05). In summary, LHe as cryogen, the supplementation of 17.5% EG and 17.5% DMSO cryoprotectant in the vitrification solution could reduce cryoinjury caused by vitrification at some extent, and significantly improved developmental capacity in vitro of MⅡ oocytes after vitrification-thawing, which helped to optimize the vitrification procedures. This study provides a theoretical basis for the efficient vitrification of bovine MⅡ oocytes using LHe as cryogen.

Excessive use of acetaminophen (APAP) can promote liver oxidative stress and cause liver damage. Forsythiaside A (FA), as the phenylethanol glycoside active component of the Chinese herb Forsythia, may have a hepatoprotective effect. To investigate the effect of FA on APAP-induced oxidative stress, C57BL/6 mice (Mus musculus) and alpha mouse liver 12 (AML12) cells were randomly divided into 4 groups: Blank control (NC), APAP-induced liver injury group (APAP), FA control group (FA), and FA treatment group (APAP+FA). Serum and cell supernatants were analyzed for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity to determine the effective concentration of FA for treating APAP-induced liver injury. Levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), and hydrogen peroxide (H₂O₂) in mouse liver tissue were measured, and pathological changes in liver tissue were observed to explore the therapeutic effect of FA on APAP-induced liver injury. A pregnane X receptor (PXR) overexpression plasmid was constructed, and C57BL/6 mice and AML12 cells were divided into 5 groups: normal control (NC), APAP-induced liver injury (APAP), APAP+PXR overexpression (APAP+PXR(OE)), FA treatment (APAP+FA), and FA treatment with PXR overexpression (APAP+FA+PXR(OE)). The effects of FA on SOD activity, GSH, MDA, H₂O₂, tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) content, PXR protein and gene expression levels, and liver pathology were observed, and a rescue experiment was designed to verify the potential mechanism. The results showed that FA significantly reduced ALT and AST activity, as well as MDA and H₂O₂ content in APAP model mice (P<0.05), increased SOD activity and GSH content (P<0.05), and improved liver tissue pathology. FA also significantly reduced MDA, H₂O₂ content, and TNF-α, IL-1β, IL-6 levels (P<0.05), and increased SOD activity and GSH content (P<0.05) by inhibiting PXR. In conclusion, FA ameliorated oxidative stress by inhibiting PXR, thereby exerting a protective effect against APAP-induced liver injury. This study provides new ideas for the treatment of drug-induced liver injury and a theoretical foundation for the hepatoprotective effect of FA through inhibiting PXR.

As a crucial cellular matrix in influenza vaccine production, MDCK (Madin-Darby Canine Kidney) cells' potential tumorigenicity remains a key factor constraining vaccine safety. Double cortin-like kinase 1 (DCLK1), a tumor stem cell marker, is highly expressed in diverse malignancies and participates in malignant progression, though its role in MDCK cell tumorigenicity remains unclear. Our research group proteomic studies identified differentially expressed protein DCLK1 between high-tumorigenic and non-tumorigenic MDCK cell lines. This study established stably DCLK1-low and overexpressed MDCK cell lines using lentiviral vector-mediated gene transfection. The effects of DCLK1 on multiple biological characteristics of MDCK cells were evaluated through CCK-8 assays, scratch assay, Transwell migration, and clonation formation experiments. Results showed that DCLK1 knockdown had no significant impact on cell morphology or proliferation capacity, but significantly reduced cell migration, invasion, and low-density proliferation capabilities. Conversely, overexpression enhanced these abilities. Further investigation revealed that DCLK1 knockdown downregulated tumor-associated factors including uroplakin 3A (UPK3A), while upregulating hepatoprotein B receptor 2 (ephrin type-B receptor 2, EphB2) and ataxin-1 (ATAXN1). DCLK1 overexpression showed no significant effects on these factors. Furthermore, constructing tumor-prone MDCK cell models in BALB/c nude mice (Mus musculus) and performing staining analysis on liver tissues revealed that vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule 1 (VCAM-1) expression showed no clear correlation with tumor development in MDCK cells. This suggested their tumorigenicity may not be regulated by these 2 factors. The study demonstrated that DCLK1 might participate in the tumorigenic process of MDCK cells by regulating downstream target gene expression. This study provides crucial insights into understanding the mechanisms underlying MDCK cell tumorigenesis, identify potential therapeutic targets for future research on reducing tumorigenicity, and hold significant implications for enhancing vaccine safety in MDCK cell applications.

As a vital member of the human recombination Q (RecQ) helicase family, Bloom syndrome mutated protein (BLM) plays essential roles in multiple cellular metabolic processes, including DNA replication, repair, recombination, transcription, and telomere maintenance. Structural studies on BLM helicase have revealed that, in addition to the 3 canonical conserved domains characteristic of the RecQ helicase family—the helicase domain (HD), RecQ C-terminal domain (RecQ-Ct) and helicase and RNase D C-terminal domain (HRDC)—a nuclear localization signal (NLS) is also identified at its carboxyl terminus (C-terminus).However, research on its amino terminus (N-terminus) remains limited. This research group's structural analysis of the BLM N-terminus has found that a large number of threonine phosphorylation sites were present in the region spanning 619~843 bp. It was hypothesized that these sites play a critical function in the post-translational modifications (PTMs) of BLM helicase, yet the cellular proteins involved in this physiological process remain incompletely elucidated. To screen for cellular proteins that interact with the N-terminal 301~650 peptide fragment of BLM helicase (BLM_301~650) and further characterize their interaction with the BLM N-terminus, this present study employed immunoprecipitation-high-performance liquid chromatography tandem mass spectrometry (IP-HPLC-MS/MS) to identify BLM_301~650-interacting proteins in the prostate cancer (PC3) cell line. Bioinformatic analyses of the screened interacting proteins were performed using the database for annotation, visualization and integrated discovery (DAVID) and search tool for the retrieval of interacting genes/proteins (STRING). The subcellular colocalization between BLM_301~650 and its interacting proteins was validated via immunofluorescence assay, while the interaction relationship was confirmed by Pull-down assays. IP-HPLC-MS/MS results demonstrated that a total of 172 proteins specifically binding to BLM_301~650 were identified. KEGG pathway enrichment analysis indicated that these interacting proteins were predominantly enriched in the ribosome pathway. Protein-protein interaction (PPI) network analysis further identified ribosomal protein S27a (RPS27A) as a key interacting factor of BLM_301~650. Immunofluorescence and Pull-down assays verified that BLM_301~650 and RPS27A colocalized in the nucleoplasm and exhibited a direct targeting relationship. Collectively, the direct interaction between RPS27A and the N?terminal region of BLM helicase serves as a novel molecular bridge for the crosstalk between the ribosomal pathway and the DNA repair pathway, and is of great scientific significance for elucidating the mechanisms underlying genome stability maintenance, tumorigenesis, and DNA damage response.

As a typical economic bivalve, inhabiting intertidal mudflats, the blood clam (Tegillarca granosa) is prone to selectively accumulate the heavy metal cadmium (Cd). Sodium calcium exchangers (NCXs) are crucial ion transporters on the cell membrane. To explore the role of T. granosa sodium calcium exchanger 1 (TgNCX1) during the selective Cd enrichment process, the blood clams were exposed to 0.089 μmol/L Cd & Cu stress for 120 h. The enrichment characteristic of Cd in T. granosa was detected by inductively coupled plasma-mass spectrometry (ICP-MS), then the sequence features of TgNCX1 were analyzed by the bioinformatics approaches, and its expression profiles were performed by qPCR and Western blot techniques. The results showed that the full-length cDNA of TgNCX1 (GenBank No. PQ255942.1) was 3 001 bp, containing an open reading frame of 2 409 bp and encoding 802 amino acids. Sequence analysis revealed that TgNCX1 belonged to the Calcium/Cation antiporter (CaCA) superfamily, and was a hydrophilic membrane protein with 9 transmembrane domains and 3 Ca²⁺-binding sites, indicating its capability of binding and transporting Ca²⁺. Tissue-specific expression analysis showed that TgNCX1 had the highest expression in the gills of T. granosa, suggesting that gill may serve as a critical channel for Cd²⁺ entry into the organism. The results of temporal expression showed that the expression of TgNCX1 gene and its encoded protein in the gills first increased, and subsequently decreased under Cd & Cu stress. During the early stage of the metal stress, the expression of TgNCX1 gene and its encoded protein increased significantly and peaked at 24 h, which was consistent with Cd enrichment pattern. It was presumed that the reverse transport mode of TgNCX1 may be triggered by Cd²⁺ competitively binding to the Ca²⁺-binding domain, driving Cd²⁺ influx through the electrochemical gradient of Na⁺, thereby mediating the short-term selective Cd enrichment in T. granosa. After 24 h of Cd & Cu stress, the expression level of TgNCX1 and its encoded protein decreased significantly, while Cd content increased markedly, suggesting that TgNCX1 may not be involved in the long-term selective Cd enrichment of T. granosa, and this process may rely on other transporters or ion channel proteins. This study further reveals the intrinsic regulatory mechanism of selective Cd enrichment, and provide a theoretical basis for cultivating new varieties with low Cd accumulation in T. granosa.

Soil microorganisms play a pivotal role in biogeochemical cycles. Understanding the evolution of research paradigms in this field is essential to advancing ecological theory and facilitating sustainable agriculture. To delineate the research trajectory and emerging frontiers of soil microbiology, this study compiled a comprehensive dataset encompassing 28 000 Chinese and English publications (1999~2024) from the WOS (Web of Science) core collection and China National Knowledge Infrastructure (CNKI). Using multi-metric analysis and knowledge mapping tools (VOSviewer and CiteSpace), keyword co-occurrence clustering, burst detection, and timeline analysis were conducted. The results indicated the following: (1) Global knowledge output in this field expanded substantially, with English-language literature growing at an average annual rate of 6.85%, while Chinese publications experienced accelerated growth since 2020. (2) Research hotspots clustered around 5 major thematic areas: Nutrient cycling, pollution remediation, community characteristics, environmental dynamics, and climate regulation. (3) Keyword burst analysis identified a three-stage paradigm shift: An initial focus on diversity characterization (1999~2010), a transition to metagenome-driven exploration (2011~2018), and a recent emphasis on interaction networks and multifunctional mechanisms (2019~2024). Future research should prioritize multi-scale dynamic monitoring of soil microbiomes, the mechanisms of functional redundancy under global change, and AI-enabled modeling of microbial interaction networks. The developed "hotspot-trend-frontier" analytical framework provides theoretical and methodological support for optimizing research strategies in soil microbiology and guiding ecological restoration practices.

Soil cadmium (Cd) contamination has become an increasingly serious environmental problem under intensified industrial and agricultural activities, posing significant threats to crop production and ecological security. Microbial remediation has attracted considerable attention as an environmentally friendly and cost-effective strategy. Synthetic microbial communities (SynComs), which are artificially assembled microbial consortia composed of functionally defined strains, represent controllable microbial systems capable of performing complex ecological functions through cooperative interactions. In recent years, SynComs have emerged as powerful tools for investigating plant-microbe interactions and for developing microbiome-based environmental solutions. This review summarizes the principles and design strategies for constructing SynComs and outlines the main approaches for their optimization and functional regulation. In addition, the potential roles of SynComs in plant-microbe interaction studies and environmental management are discussed, with a particular focus on their application in enhancing plant tolerance to cadmium stress. Overall, this review aims to provide theoretical insights into the ecological and molecular mechanisms underlying SynCom-mediated plant tolerance to heavy metal stress, and to offer new perspectives for the application of SynComs in the remediation of heavy metal-contaminated soils and sustainable agricultural development.

Shikonin, a highly liposoluble naphthoquinone compound isolated from the dried roots of Lithospermum erythrorhizon, has been demonstrated to possess multiple biological functions, including antimicrobial, anti-inflammatory, anti-heat stress, and wound-healing promoting activities. Given its unique bioactive properties, this compound exhibits significant application potential in green antibiotic-free fields such as alleviating transport stress in livestock and poultry, promoting animal wound repair, developing feed additives, advancing the development of antibiotic-free feed, and food preservation. Therefore, this article comprehensively reviewed the mechanisms of action of shikonin against pathogenic microorganisms and summarized its potential applications and research findings in livestock production, including its role in alleviating transport stress, promoting wound repair, and serving as a feed additive. The review provides a reference for the development and utilization of shikonin in animal production.

Aeonium arboreum and its cultivars, A. arboreum and its range of succulent cultivars in the genus Aeonium of the Crassulaceae family, are important indoor ornamental plants. The absence of a transformation system in this kind of plant restricts its molecular-level studies. This study utilized the A. arboreum cultivars A. arboreum 'Halloween', A. arboreum 'Velour European' and A. arboreum as materials. Different whorl leaves and different strains of Agrobacterium rhizogenes were used to produce hairy roots under non-tissue cultured conditions, aiming to establish a non-tissue cultured hairy root transformation system for A. arboreum and its cultivars. The results indicated that using intermediate whorl leaves and the Agrobacterium rhizogenes strain ArQual could achieve the highest hairy root induction rates, with hairy root induction rates of 93.33%, 96.67% and 83.33% for A. arboreum 'Halloween', A. arboreum 'Velour European' and A. arboreum, respectively. After infection with ArQual carrying the pBI121 vector, the hairy roots produced by the intermediate whorl leaves of the A. arboreum and its cultivars turned blue upon GUS staining. The transformation efficiency of hairy root from A. arboreum 'Halloween', A. arboreum 'Velour European' and A. arboreum were 89.93%, 82.22% and 69.80%, respectively. The anthocyanin-regulating transcription factor AaMYB113 from A. arboreum 'Halloween' was used to validate the hairy roots transformation system of the A. arboreum and its cultivars, and the results showed that compared with the hairy roots transformed with an empty vector, the hairy roots transformed with AaMYB113 exhibited significantly redder coloration. Both anthocyanin content and the expression levels of anthocyanin biosynthetic structural genes in 'Halloween' hairy roots were significantly increased (P<0.05), demonstrating the feasibility of this transformation system. This study established a non-tissue cultured transformation system of hairy roots in A. arboreum and its cultivars, providing technical support for subsequent functional gene research and molecular breeding in these plants.

Metal-organic frameworks (MOFs) represent a class of emerging porous crystalline materials constructed from metal ions serving as nodes and organic ligands acting as linkers. They exhibit high porosity, extensive specific surface area, and excellent biocompatibility, rendering them suitable for diverse applications in biomedical fields, gas storage, trace element detection, and sensing. Through the strategic design of specific metal ion and organic ligand combinations, MOFs can generate efficient photodynamic effects, thereby garnering considerable research interest in antibacterial and antitumor applications. In this study, silver metal-organic framework (Ag-BTC) was synthesized via a solvothermal method employing silver nitrate (AgNO₃) and 1,3,5-benzenetricarboxylic acid (BTC) as precursors. The physicochemical properties of Ag-BTC were characterized. Its light-induced reactive oxygen species (ROS) generation capacity and pH-responsive behavior were investigated using electron paramagnetic resonance (EPR) and inductively coupled plasma-optical emission spectrometry (ICP-OES). Antibacterial activity against Staphylococcus pseudintermedius was evaluated via the spread plate method and scanning electron microscopy (SEM). The results demonstrate that, under simulated sunlight irradiation for 10 minutes, Ag-BTC simultaneously produced singlet oxygen (¹O₂) and hydroxyl radicals (·OH). Notably, its decomposition efficiency in weakly acidic conditions was approximately two-fold higher compared to neutral environments, indicating pronounced pH-responsive characteristics. The antibacterial rate against S. pseudintermedius reached 97.4%, inducing bacterial membrane shrinkage, perforation, and DNA leakage, corroborating its significant antibacterial effects. This study provides a theoretical foundation for the potential application of Ag-BTC in the veterinary clinical treatment of bacterial skin infections.