BACKGROUND: Ovarian cancer remains a therapeutic challenge due to high recurrence rates and chemoresistance, necessitating novel agents from natural sources. In our previous work, a triterpenoid saponin Raddeanoside R7 (R7) was isolated and characterized from Pulsatilla cernua (Thunb.) Bercht. ex J. Presl (P. cernua). Here, R7 was investigated for its anti-ovarian cancer potential. METHODS: The cytotoxic effects of R7 on Caov3 and OVCAR3 cells were evaluated using the CCK-8 assay. Cell migration was assessed via the Transwell migration assay. Flow cytometry was employed to analyze cell cycle distribution and apoptosis. The role of reactive oxygen species (ROS) production in R7’s inhibitory effects was investigated through fluorometric analysis using the DCFH-DA probe. The underlying mechanisms were further explored through integrated transcriptomics and network pharmacology, with core targets validated by molecular docking. The effect of R7 on underlying signaling pathway was examined by western blotting. RESULTS: R7 exhibited potent cytotoxicity against Caov3 and OVCAR3 cells, with half-maximal inhibitory concentrations (IC50) values of 7.27 µM and 8.25 µM, leading to significant inhibition of cell viability. The Transwell migration assay showed that R7 treatment reduced the number of migrated cells by over 60%. Flow cytometry analysis revealed that R7 induced S phase cell cycle arrest and promoted apoptosis, with early and late apoptosis rates increasing to a total of approximately 68% and 29.9% of Caov3 and OCVAR3 cells, respectively. Furthermore, ROS production was identified as a key mediator of these inhibitory effects. Integrated analysis identified CCND2 and HSPA8 as core targets, which was confirmed by molecular docking. Counterintuitively, activation of the PI3K-AKT pathway was found to potentially mediate the therapeutic effects of R7. CONCLUSIONS: R7 acts as a novel agent against ovarian cancer by inhibiting proliferation and migration, inducing S phase arrest and apoptosis, and elevating ROS levels. Its anti-tumor effects are potentially mediated through a context-dependent activation of the PI3K-AKT pathway, nominating R7 as a promising therapeutic candidate for further investigation.

Pulsatilla chinensis (Bge.) Regel is a widely used Chinese medicinal herb and is prone to adulteration because of morphological similarity among congeneric species. It shows substantial quality variations across Northeast, North, and East China. This study combined hyperspectral imaging (HSI) with three feature-wavelength selection algorithms, including competitive adaptive reweighted sampling (CARS), successive projections algorithm (SPA), and uninformative variable elimination (UVE), to identify eight Pulsatilla species and trace the geographical origins of P. chinensis from three major producing areas. Six classifiers were compared, including support vector machine (SVM), multilayer perceptron (MLP), gated recurrent unit (GRU), convolutional neural network (CNN), Transformer, and a CNN+Transformer hybrid. Spectral differences among species and origins were detected in bands associated with chemical constituents, such as proteins. Deep learning models outperformed conventional machine learning. For species identification, the CNN+Transformer hybrid with CARS achieved the highest accuracy of 85.41%, reduced the spectral dimensionality by 65.9%, and improved the accuracy by 7.15% over the full-spectrum input. For geographical origin tracing, the same hybrid reached 88.58% accuracy using full-spectrum data. These results indicate that HSI integrated with deep learning enables rapid, nondestructive species identification and geographical origin tracing of Pulsatilla for quality assessment of traditional Chinese medicine.

Pulsatilla chinensis (Bge.) Regel is a widely used Chinese medicinal herb and is prone to adulteration because of morphological similarity among congeneric species. It shows substantial quality variations across Northeast, North, and East China. This study combined hyperspectral imaging (HSI) with three feature-wavelength selection algorithms, including competitive adaptive reweighted sampling (CARS), successive projections algorithm (SPA), and uninformative variable elimination (UVE), to identify eight Pulsatilla species and trace the geographical origins of P. chinensis from three major producing areas. Six classifiers were compared, including support vector machine (SVM), multilayer perceptron (MLP), gated recurrent unit (GRU), convolutional neural network (CNN), Transformer, and a CNN + Transformer hybrid. Spectral differences among species and origins were detected in bands associated with chemical constituents. Deep learning models outperformed conventional machine learning. For species identification, the CNN + Transformer hybrid with CARS achieved the highest accuracy of 85.41%, reduced the spectral dimensionality by 65.9%, and improved the accuracy by 7.15% over the full-spectrum input. For geographical origin tracing, the same hybrid reached 88.58% accuracy using full-spectrum data. These results indicate that HSI integrated with deep learning (CNN + Transformer) enables rapid, nondestructive species identification and geographical origin tracing of Pulsatilla for quality assessment of Traditional Chinese Medicine.

This study sequenced and assembled the complete chloroplast genome of Adenophora pinifolia via next-generation sequencing. The genome is 166,976 bp, with a 121,189 bp LSC, 26,959 bp SSC, and two 9,414 bp IR regions. Its total GC content is 38.33%. Annotation revealed 127 genes: 74 protein-coding, 8 rRNA, 38 tRNA, and 7 pseudogenes. Maximum likelihood phylogenetic analysis showed A. pinifolia is closely related to Adenophora erecta and Adenophora remotiflora, forming a monophyletic clade. These data support its phylogenetic research and resource utilization.

Anemone sylvestris Linnaeus (1753) is a species in the Ranunculaceae family within the genus Anemone. It is mainly distributed in China, Croatia, and Poland. The chloroplast genome has a total length of 161,033 bp, comprising a large single-copy region of 81,144 bp, a small single-copy region of 17,393 bp, and two inverted repeat regions of 31,248 bp each. The GC content is 37.54%. A total of 134 genes were annotated, including 90 protein-coding genes, 8 rRNA genes, and 36 tRNA genes. Phylogenetic analysis using the maximum likelihood method robustly supported the close relationship of A. sylvestris with other Anemone species, namely A. raddeana, A. reflexa, and A. altaica, forming a well-defined monophyletic clade. The complete chloroplast genome of A. sylvestris reported here provides a useful resource for subsequent phylogenetic, identification, and resource related studies.

Cytoplasmic male sterility (CMS), a maternally inherited trait, is a valuable approach employed for heterosis utilization in plant breeding programs. Radish is an important vegetable crop that exhibits significant heterosis. The utilization of CMS lines as female parents is essential for efficient F1 hybrid seed production in radish. Among various CMS types, Ogura CMS is widely used, and its molecular mechanism has been extensively studied in radish and other Brassica crops, including cabbage and broccoli. However, the molecular modulator and underlying regulatory mechanism of DCGMS in radish remain elusive. In this study, six radish genotypes were identified as DCGMS with molecular markers based on the male sterility gene orf463. Microspore abortion of DCGMS line appeared shortly after the tetrad stage, when the tapetal cells grew abnormally large. Based on transcriptome data and bioinformatic analyses, RsMYB3R-1 was identified as being highly expressed in stamens, and its encoded protein was predicted to target mitochondria. Yeast one-hybrid and dual-luciferase assays demonstrated that RsMYB3R-1 bound to the orf463 promoter. Transient overexpression of RsMYB3R-1 in radish cotyledons could increase the expression level of orf463. This result may cause mitochondrial energy production dysfunction and abnormal pollen development in the DCGMS lines. Taken together, our findings elucidate that the RsMYB3R-1-orf463 module plays a pivotal role in mediating pollen abortion in the DCGMS line of radish. These results not only offer novel insights into the nucleus–mitochondria interaction, but also establish a foundation for deciphering the molecular mechanisms underlying DCGMS male sterility, thereby facilitating the efficient utilization of heterosis in radish breeding.

Radish (Raphanus sativus L.) is a cool-season root vegetable crop grown worldwide. Heat stress (HS) severely restricts radish taproot formation, resulting in the loss of yield and quality. However, the regulatory mechanism underlying radish taproot thickness under HS conditions has been largely unexplored. Here, we identified a heat-induced HSP protein, HEAT SHOCK PROTEIN 22 (RsHSP22), via comparative proteome analysis. Yeast one-hybrid (Y1H) assays, dual-luciferase reporter assays, and electrophoretic mobility shift assays demonstrated that WRKY DNA-BINDING PROTEIN 18 (RsWRKY18) and HEAT STRESS TRANSCRIPTION FACTOR A2 (RsHSFA2) bind to the W-box and HSE element of the promoter of RsHSP22 to activate its expression, respectively. RsWRKY18 and RsHSFA2 are heat-inducible and nucleus-localized transcription activators, and their expression levels in the heat-tolerant genotype "NAU-XBC" were higher compared to the heat-susceptible genotype "NAU-YB" after a short- or long-term HS treatment at the taproot thickening stage. Overexpression of RsWRKY18 and RsHSFA2 alleviated oxidative damage under HS and conferred heat tolerance in Arabidopsis (Arabidopsis thaliana) and radish, whereas silencing RsWRKY18 resulted in heat susceptibility and limited taproot thickening in radish. Furthermore, RsWRKY18 bound to the promoters of several taproot thickening-related genes, including Xyloglucan endotransglucosylase/hydrolase 32 (RsXTH32), Alpha-expansin 9 (RsEXPA9), KNOTTED-like from Arabidopsis thaliana 1 (RsKNAT1), and WUSCHEL homeobox-containing 14 (RsWOX14). Interestingly, RsWRKY18 interacted with RsHSFA2 coordinately to enhance the transactivation activity of both RsWRKY18 and RsHSFA2 on their target genes, especially under HS conditions. Overall, these results show that the module RsWRKY18-RsHSFA2 regulates the HS response and taproot thickening in radish and helps facilitate the development of heat-tolerant cultivars with superior yield and quality in radish and other root vegetables.

Cadmium (Cd), a non-essential and toxic heavy metal, accumulates in radish taproot and adversely affects radish safety and quality. Pleiotropic drug resistance (PDR) proteins play a crucial role in heavy metal uptake and accumulation in plants. However, the PDR gene-mediated regulation of Cd accumulation remains unexplored in radish. RsPDR12 gene expression significantly increased under Cd stress. RsPDR12 demonstrated Cd transport activity and decreased Cd accumulation in yeast cells, and its overexpression reduced root Cd concentration by Cd2+ efflux in Arabidopsis. RsPDR12 alleviated Cd stress by enhancing membrane permeability and reactive oxygen species (ROS) scavenging in radish plants. RsWRKY15 was identified as the upstream regulatory factor of RsPDR12. Dual-luciferase assay demonstrated that RsWRKY15 bound to the RsPDR12 promoter to activate its transcription. RsWRKY15 expression and promoter activity were significantly induced under Cd stress. RsWRKY15 overexpression mitigated oxidative damage and reduced root Cd concentration in radish and Nicotiana benthamiana plants, respectively. These findings advance understanding of the molecular mechanism underlying the RsWRKY15–RsPDR12 mediated regulatory network of Cd accumulation and support the genetic improvement of low Cd-accumulation cultivars in radish breeding programs.

The DMP (Domain of Unknown Function 679 Membrane Proteins) gene family consists of proteins specifically expressed in plant membranes with functions associated with gamete fusion. However, its functional characterization remains largely unexplored in radishes (Raphanus sativus L.). In this study, a total of 13 DMP genes were identified in the radish genome; they were distributed unevenly across six chromosomes. Phylogenetic analysis revealed five distinct clades, based on the classification and nomenclature of this gene family in Arabidopsis. RsDMP9 was grouped with the haploid-inducing genes AtDMP8 and AtDMP9, which play a potential role in haploid induction. Light, growth, hormone, and stress-responsive elements were identified in the promoter of RsDMP9. Quantitative real-time polymerase chain reaction demonstrated the preferential expression of RsDMP9 in pollen, while other RsDMP genes showed differential expression in the roots, petals, and sepals. The RsDMP9 protein was confirmed to be localized to the plasma membrane. The identification and cloning of endogenous U6 promoter sequences from the radish genome were achieved based on the conserved U6 small nuclear RNA (snRNA) sequences of Arabidopsis thaliana. In vivo imaging and a dual-luciferase reporter system indicated that RsU6-6 exhibited strong transcriptional activity comparable to that of AtU6-1; truncation of RsU6-6 to 326 base pairs could enhance transcriptional activity, making it ideal for single guide RNA (sgRNA) expression and multiplex editing by avoiding promoter crosstalk. Additionally, the CRISPR/Cas9 system driven by RsU6-6 has been shown to achieve a high mutation frequency by protoplast transformation technology, thus demonstrating the effective validation of the CRISPR/Cas9 editing vector in radish protoplasts. These findings provide insights into the evolutionary conservation and functional diversification of RsDMP genes, and this protoplast-based editing system would facilitate effectively validating gene function and precision improvement of important traits in radish breeding programs.

Protoplasts serve as a versatile platform for genetic transformation and somatic hybridization. Radish (Raphanus sativus L.) is an important root vegetable crop, and its taproot is derived mainly from the hypocotyl and main root. However, an efficient protoplast isolation and transient transformation system for radish, particularly for hypocotyl tissues, has not been well established. In this study, we developed an optimized, tissue-specific protocol for protoplast isolation and transformation from radish cotyledon and hypocotyl. Adding 0.4% polyvinylpyrrolidone (PVP) to the enzyme solution significantly improved protoplast yield and viability while reducing intracellular reactive oxygen species (ROS) accumulation. Under optimized conditions, cotyledon-derived protoplasts (CDPs) achieved a yield of 3.28 × 106 protoplasts g-1 fresh weight (FW) with 95.4% viability and 76.62% transformation efficiency, whereas hypocotyl-derived protoplasts (HDPs) reached 1.48 × 106 protoplasts g-1 FW with 94.3% viability and 69.33% transformation efficiency. The protocol was effective across radish genotypes and several Brassicaceae root crops. Furthermore, the resulting protoplasts supported subcellular localization and gene function assays, and rapid assessment of the editing efficiency of CRISPR/Cas9 vectors carrying different Cas9 promoters and single-guide RNAs. Overall, this system would provide a practical tool for gene function analysis and precision breeding in radish and related root vegetable crops.

Conventional methods for analyzing monosaccharides, such as 1-Phenyl-3-methyl-5-pyrazolone (PMP) derivatization coupled with high-performance liquid chromatography (HPLC), struggle to detect ketoses and acidic sugars. This limitation hinders the precise structural analysis of polysaccharides in traditional Chinese medicine (TCM). A methylation-assisted Gas Chromatography-Mass Spectrometry (GC-MS) approach was developed to address this issue in the current study. By optimizing the methylation process and using GC-MS with a BR-17 column and an Electron Ionization (EI) source, simultaneous detection of ten different monosaccharides was achieved within 20 min. GC-MS profiling of monosaccharides generally gives two peaks per analyte, except in the cases of fructose and galacturonic acid. Notably, fructose (Fru) was identified for the first time in the polysaccharides of Polygonatum species: P. sibiricum at 18.55 μg/mg, P. cyrtonema at 16.90 μg/mg, and P. kingianum at 14.35 μg/mg, overcoming the limitations of the inability to detect ketoses in the PMP-HPLC method. Data analysis revealed distinct species-specific markers: P. kingianum polysaccharides (PKP) uniquely contained α-galacturonic acid (α-Gal: 19.95 μg/mg), whereas P. sibiricum polysaccharides (PSP) featured six monosaccharides, including glucose (Glc: 135.45 μg/mg), mannose (Man: 78.95 μg/mg), and ribose (Rib: 26.90 μg/mg), with a total of 299.50 μg/mg. P. cyrtonema polysaccharides (PCP) showed elevated levels of glucuronic acid (GlcA, 38.40 μg/mg). This derivatization-enhanced GC-MS method effectively fingerprints TCM polysaccharides, enabling clear differentiation of Polygonatum species through analysis of monosaccharide composition and total sugar content. This approach establishes clear structural distinctions, overcoming the limitations of conventional chromatographic techniques and enhancing the quality control and authentication of TCM polysaccharides.

IntroductionBlechnopsis orientalis (L.) C. Presl is a medicinal and edible fern species belonging to the Blechnaceae family. Currently, the complete mitochondrial genome of B. orientalis, as well as those of other Blechnaceae species, remains unreported, and studies on fern mitochondrial genome are limited.MethodsIn this study, the B. orientalis mitochondrial genome was sequenced using both Nanopore PromethION and Illumina NovaSeq 6000 platforms. Genome annotation was performed using MITOFY and MFANNOT, with structural visualization via OGDRAW. In-depth analyses were conducted, including assessments of non-synonymous/synonymous mutation ratios (Ka/Ks), codon usage bias, repeat sequence identification, RNA editing site prediction, collinearity, and the identification of homologous fragments between chloroplast and mitochondrial genomes. Finally, we employed both the maximum likelihood (ML) and Bayesian (BI) methods to analyze the phylogenetic relationships among B. orientalis and nine other fern and lycophyte species.ResultsThe mitochondrial genome of B. orientalis has a complex structure comprising 80 contigs, with a total length of 501,663 bp and a GC content of 48.53%. A total of 179 genes were identified, including 40 protein-coding genes (PCGs), 98 tRNA genes, 40 rRNA genes, and one pseudogene (rps11). Phylogenetic analysis based on PCGs from both chloroplast genome and mitochondrial genome aligned with the relationships described in the Pteridophyte Phylogeny Group I (PPG I) system. Further comparison with mitochondrial genome of ten other reported fern and lycophyte species revealed that the mitochondrial genome PCGs in these plants are highly conserved, despite significant genome rearrangements among mitochondrial genome.DiscussionThe findings of this study provide valuable insights into the evolutionary analysis of B. orientalis and contribute to understanding the characteristics and evolutionary relationships of mitochondrial genome in ferns and lycophytes.

Salt stress is a major environmental factor limiting the production and quality of plants worldwide. Radish (Raphanus sativus L.), one of the most important root crops, is susceptible to salt stress worldwide. Plasma membrane intrinsic proteins (PIPs) have been identified to play a crucial role in regulating plants' salt tolerance. However, the underlying molecular regulatory mechanisms involved in salt stress tolerance are largely unknown. Here, a salt-induced water transport gene RsPIP2-1 associated with the regulatory mechanisms in response to salt stress was clarified in radish. Overexpression of RsPIP2-1 had high-water channel and H2O2 transport activity in Xenopus laevis oocytes and yeast, and it also conferred prominently salt tolerance through promoting reactive oxygen species (ROS) scavenging and enhancing antioxidant enzyme activity in transgenic radish. Moreover, yeast one-hybrid (Y1H) was used to screen the upstream regulators of RsPIP2-1, and two ethylene-responsive transcription factors including RsCBF2 and RsERF18 were identified. Y1H, dual-luciferase assay (DLA) and electrophoretic mobility shift assays (EMSA) showed that these two genes could active the transcription of RsPIP2-1 by directly binding to the DRE/CRT element and GCC-box element in its promoter. In addition, the salt tolerance and the expression levels of these two transcription factors could be significantly upregulated when treated with exogenous application of an ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), while the plants' resistance as well as the expression patterns could be reduced when exposure to the inhibitor of ethylene action (AgNO3), suggesting that RsCBF2 and RsERF18 positively regulated the salt tolerance in a manner of dependent on ethylene synthesis pathway. Taken together, these findings uncover a novel transcriptional regulatory module based on the RsCBF2/RsERF18-RsPIP2-1 underlying salt tolerance in radish and could provide new insights into the salt-tolerant vegetable crop breeding programs.

Acting as a nucleus-localized transcriptional activator, RsWRKY75 promotes ROS scavenging and Cd efflux by activating the transcription of RsAPX1 and RsPDR8 in radish. Radish (Raphanus sativus L.) is an important economical root vegetable crop worldwide. As a toxic heavy metal, cadmium (Cd) can dramatically hamper radish taproot quality as well as threaten human health. Although the WRKY transcription factors (TFs) play crucial roles in plant response to Cd stress, how WRKY TFs mediate Cd uptake and efflux remains elusive in radish. Herein, the RsWRKY75, belonging to the WRKY-IIc sub-group, displayed high expression in vascular cambium at the expanding stage, whose promoter activity and expression were obviously induced by Cd exposure at 24 h in radish root. RsWRKY75 was localized primarily to the nucleus and had transactivation activity in yeast and tobacco leaf cells. Transient transformation indicated that RsWRKY75 promoted Cd-induced ROS scavenging in radish cotyledons. Overexpression of RsWRKY75 led to increased root elongation but decreased Cd accumulation in Arabidopsis plants. Both in vitro and in vivo assays revealed that RsWRKY75 bound to the RsAPX1 promoter and activated its expression to eliminate excessive ROS accumulation. Moreover, RsWRKY75 activated RsPDR8 transcription by directly binding to its promoter, thereby promoting Cd efflux in the radish root. Collectively, we revealed a novel module of RsWRKY75-mediated ROS scavenging and Cd efflux in radish. These results would facilitate to establish genetic strategies to achieve RsWRKY75-dependent Cd extrusion and detoxification in radish.

Cold stress adversely affects crop growth and development. WRKY transcription factors play the critical role in regulating abiotic stress response and balancing plant growth and cold stress defense. However, the roles of WRKY in the trade-off between root growth and cold stress response are still rarely known in radish. Herein, RsWRKY49 was specifically expressed in the radish root and the expression was highly induced by low temperature in the root of cold-tolerant radish genotype 'NAU-RG' compared to the cold-sensitive radish genotype 'NAU-XBC'. Overexpression of RsWRKY49 in 'NAU-XBC' enhanced cold tolerance, while interference of its expression in 'NAU-RG' increased cold sensitivity. The increase in cell division activity and root meristem size was observed in the radish hairy root overexpressing RsWRKY49 under both normal and low-temperature conditions, demonstrating its ability to regulate cold stress response and root growth. Natural variation in the RsWRKY49 promoter affects the differences in its expression level in different cold-tolerant radish genotypes, thereby modulating cold tolerance. Comparative promoter analysis identified additional cis-acting regulatory elements (ten TATA boxes, two ABRE elements, and one DRE element) in the RsWRKY49 promoter of 'NAU-RG', which showed enhanced promoter activity compared to that of 'NAU-XBC' under cold stress. In addition, RsWRKY49 could transactivate RsCBF2 and RsNR2 expression to regulate cold stress response. These results provide insights into the molecular mechanism underlying WRKY TFs balancing root growth and defense to cold stress in radish and would facilitate achieving genetic improvement of cold-tolerant cultivars in radish breeding programs.

Spring-type Brassica rapa L. is a valuable genetic resource for breeding early-maturing crops, offering advantages such as early flowering and rapid maturation. However, the genetic mechanisms governing flowering time (FT) in spring-type B. rapa remain insufficiently understood. In this study, we investigated the flowering-time trait of an extremely early-maturing landrace, “Haoyou 11”, originating from the Qinghai-Tibetan Plateau. Initial mapping was conducted using an F2 population derived from the cross between Haoyou 11 and Dahuang (a late-flowering spring-type landrace of B. rapa). A major quantitative trait locus (QTL) for flowering time, designated qFTA06, was identified within a 1.70 Mb interval on chromosome A06 using genotyping-by-sequencing (GBS) and bulked segregant analysis sequencing (BSA-seq). The locus qFTA06 was subsequently fine-mapped to a 75.16 kb region with a set of near-isogenic lines (NILs), and BrCDF3, a gene encoding a Dof transcription factor, was identified as the causal gene underlying qFTA06. Virus-induced gene silencing (VIGS) experiments revealed that BrCDF3 acts as a negative regulator of flowering time under long-day (LD) conditions, with sequence variation contributing to the early-flowering phenotype in Haoyou 11. Phenotypic analysis of NILs showed that NIL-E, carrying the BrCDF3 allele from Haoyou 11, flowered approximately 7 days earlier than NIL-L, which harbors the BrCDF3 allele from Dahuang. By employing CRISPR/Cas9 technology, we further validated that the homologous gene BnCDF3 also functions as a negative regulator of flowering time in Brassica napus L., and analyzed natural variations in the CDF3 gene across natural populations. This study provides new insights into the genetic basis of flowering time in spring-type B. rapa, advancing early-maturity breeding efforts in crops.

The lung, a fragile yet crucial organ for breathing in humans, is susceptible to harm from harmful external elements. This study advances the extraction of Arctium lappa polysaccharide (ALP) by our team, aiming to broaden the possibilities for novel medications targeting lung conditions. The structure of ALP was further analyzed by congo red analysis. Effect of ALP pre-treatment was checked by assessing antioxidant markers, inflammatory cytokines and antiapoptosis in vitro and in vivo. Congo red analysis showed that ALP had a stable three-helical conformation. ALP conferred protection against acute lung injury (ALI) induced by lipopolysaccharide (LPS) through the attenuation of lung tissue injury and inhibition of cell apoptosis. The apoptosis rate in lung tissues was markedly reduced in ALP-treated groups compared to the LPS group (P < 0.01). This protective effect is primarily attributed to dual mechanisms: the suppression of inflammatory cytokine synthesis and the enhancement of antioxidant responses. ALP significantly reduced the levels of LPS-induced inflammatory cytokines (P < 0.05) including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in vitro and in vivo experiments. In addition, ALP alleviated LPS-induced lung injury by reducing malondialdehyde (MDA) and upregulating superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and the overall antioxidant capacity (T-AOC) contents. The western blot analysis of mice lung tissues revealed that ALP pretreatment significantly attenuated the LPS-induced upregulation of key inflammatory signaling proteins (P < 0.05), including TLR4, MyD88, p65, P-p65, i-κB, p-i-κB, JNK, and p38 protein levels. The results of RAW264.7 cell experiments further veriffed that ALP inhibited TLR4/NF-κB/MAPK pathway. In conclusion, ALP can reduce LPS-induced ALI by inhibiting inflammation and oxidative stress.

Conium maculatum L. (1753) is a toxic biennial plant in the Apiaceae family. It is native to Europe and North Africa and is listed as an invasive species in China. Its 153,090 bp chloroplast genome has 132 genes (37 tRNA genes, 8 rRNA genes, and 87 protein-coding genes), structured into 2 IRs (25,087 bp), one LSC (85,716 bp) and one SSC(17,200 bp), with 37.52% GC content. Phylogenetically, it is closest to Angelica sinensis, Trachydium subnudum, Glehnia littoralis, Heracleum hemsleyanum, Peucedanum praeruptorum, and Saposhnikovia divaricata. These findings support genetic preservation of the species and Coniumphylogenetic study.

Cold stress adversely affects crop growth and development. Radish is an important root vegetable crop, and its taproot formation is susceptible to low temperatures. However, the molecular basis of the cold stress response has not yet been fully dissected in radish. Here, a sucrose phosphate synthase gene (RsSPS1) was identified through a genome-wide association study and transcriptome analysis. RsSPS1 was responsible for sucrose synthesis, and sucrose was shown to be involved in taproot growth, cambium activity, and cold tolerance in radish. RsSPS1 regulated cambium activity and cold stress response by modulating sucrose content. Moreover, RsWRKY40 was identified as the upstream transcription activator of RsSPS1 by binding to its promoter. RsWRKY40 functioned in cambium activity and cold tolerance by modulating RsSPS1-mediated sucrose accumulation. Furthermore, RsWRKY40 promoted the RsCBF1 and RsCBF2 expression levels, resulting in elevated cold resilience. RsWRKY40 also enhanced its own transcription, forming a positive auto-regulatory loop to regulate cold stress response in radish. Together, a transcription module of RsWRKY40 orchestrated cold stress response by integrating sucrose accumulation and the CBF-dependent pathway was uncovered. These findings would provide novel insight into the molecular mechanism underlying cold-responsive sucrose accumulation and cambium activity and facilitate the genetic improvement of cold tolerance in radish breeding programs.